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Biochemical, microbial and sensory evaluation of white soft cheese made from cow and lupin milk
Accepted Manuscript Biochemical, microbial and sensory evaluation of white soft cheese made from mixture cow and lupin milk Mohamed O. Elsamani , Siddig S. Habbani , Elfadil E. Babiker , Isam A. Mohamed Ahmed PII:
S0023-6438(14)00232-1
DOI:
10.1016/j.lwt.2014.04.027
Reference:
YFSTL 3863
To appear in:
LWT - Food Science and Technology
Received Date: 10 November 2011 Revised Date:
21 July 2012
Accepted Date: 13 April 2014
Please cite this article as: Elsamani, M.O., Habbani, S.S., Babiker, E.E., Mohamed Ahmed, I.A., Biochemical, microbial and sensory evaluation of white soft cheese made from mixture cow and lupin milk, LWT - Food Science and Technology (2014), doi: 10.1016/j.lwt.2014.04.027. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
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Biochemical, microbial and sensory evaluation of white
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soft cheese made from cow and lupin milk
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Mohamed O. Elsamania, Siddig S. Habbania, Elfadil E. Babikerb and Isam A. Mohamed
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Ahmedc,d* a
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Department of Food Science and Technology, Faculty of Agriculture, Omdurman Islamic University, Omdurman, Sudan
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Department of Food Science and Nutrition, College of Food and Agricultural Sciences, King Saud University, P. O. Box 2460, Riyadh 11451, Kingdom of Saudi Arabia
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Department of Food Science and Technology, Faculty of Agriculture, University of Khartoum, Khartoum North, Shambat
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Department of Applied Resources Chemistry, Faculty of Agriculture, Tottori University, Tottori 680-8553, Japan
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E-mail: [email protected]
Corresponding author: Tel.:+81 857 31 5443; Fax: +81 857 31 5347
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Abstract
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The aim of the present study was to evaluate some physicochemical, microbiological and sensory
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properties of fresh and matured (75 days) soft cheeses made with mixtures of cow milk and 0,
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25, 50 and 75 mL/100 mL of lupin milk. A remarkable increase in cheese yield was observed
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with increasing the level of lupin milk to the mixture. Compared to cow milk cheese, the protein
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content was significantly (P ≤ 0.05) increased while ash was decreased with the increase in the
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level of lupin milk for both fresh and matured cheese. However, fat content, total solids and
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acidity were increased only for fresh cheese and decreased for mature one compared to that of
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cow milk. The pH showed significant (P ≤ 0.05) reduction when the levels of lupin milk
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increased for fresh cheese while for matured cheese it slightly decreased. The total bacterial
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count is within the range that naturally exists in milk containing foods. The others
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microorganisms such as fungi, mould, E. coli, and Salmonella were not existed in both types of
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cheese. Regardless of cheese colour, incorporation of lupin milk at low concentration (25
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mL/100 mL) significantly (P ≤ 0.05) enhanced the taste, texture, flavor, and overall acceptability
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of both fresh and mature cheese.
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Keyword: Legumes, Lupinus albus, Lupin milk, sensory quality, white soft cheese
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1. Introduction Species of the genus Lupinus have been used for human nutrition for over 6000 years. Of
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them, white lupin (Lupinus albus L.) is one of the oldest agricultural crops widely used in the
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world not only as a protein source in fodder production but also for soil improvement (Huyghe,
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1997). In the past decades, the production and consumption of white lupin have increased due to
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the capability of lupin plants to grow in adverse climates, their tolerance to poor soils and their
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high crop yields (1000–2000 kg/ha) (Linnemann & Dijkstra, 2002). The protein content of lupin
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seeds ranges from 30 to 40 g/100 g with an amino acid composition comparable to that of soy
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bean. Although lupin seeds are mainly used as a feed, they are acquiring increasing importance
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as a source of vegetable proteins for human consumption in many countries. Hence, increased
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consumer awareness on health issues has produced a dramatic and fast-growing demand of plant
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protein food products. In particular, lupin proteins are receiving attention in terms of health
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benefits, particularly in relation to a number of conditions now known as ‘metabolic syndrome’
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which includes a cluster of factors such as, obesity, high blood pressure, insulin resistance and
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elevated blood cholesterol (Arnoldi, 2005). Lupin enriched foods have the potential to:
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beneficially influence satiety (appetite suppression) and energy balance (Lee et al., 2006),
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beneficially influence glycaemic control (Hall, Thomas & Johnson, 2005), improve blood lipids
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(Martins et al., 2005), reduce hypertension (Pilvi, Jauhiainen, Cheng, Mervaala, Vapaatalo, &
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Korpela, 2006) and improve bowel health (Johnson, Chua, Hall & Baxter, 2006). Today, a
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market for the use of lupin seeds in food has been developed and different products are available,
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such as pasta, bakery products containing lupin flour, meat products, beverages and tofu
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(Jayasena, Khu, & Nasar-Abbas, 2010; Paraskevopoulou, Provatidou, Tsotsiou, & Kiosseoglou,
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2010).
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Cheese is a dairy product that has played a key role in human nutrition for centuries. The
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broad range of different cheeses available is based mainly on regional conditions and production
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technology, which has been repeatedly adapted and optimized. The main objective has always
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been and still is to convert milk, which is perishable, into a product with a longer shelf life whilst
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preserving most of its nutrients (Mohamed Ahmed, Babiker, & Mori, 2010). The growth in the
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ready meals sector in recent years had been reflected in the increase in the demand for cheese as
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a food ingredient. Shredded, diced, sliced, and even liquid cheese has been developed to meet the
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needs of the modern food industry, as convenience foods continue to grow in popularity (Byrne,
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1999). The need to increase the amount of cheese from a given amount of milk is an economic
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necessity when the milk supply is limited, but the demand for such product is high. Research
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efforts have been directed mainly toward evaluating procedures for using soy bean protein
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isolate as an extender in cheeses with limited success due to either the difficulty in incorporating
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the soy protein isolate into the finished cheese or because of quality issues with the finished
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product. Seeds of other plants (such as Lupinus, reported as a possible substitute of soy in human
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foodstuffs) have not received enough attention, not because they do not contain proteins that may
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be functionally or nutritionally equal to soy bean, but rather because of economic reasons.
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However, this situation may change, and it seems advisable at present to explore the
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development and utilization of a variety of foods derived from or containing a wide range of
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plant proteins. Recent attempts have been made to process lupin seeds to produce milk-like,
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dairy replacement products, similar to those derived from soybeans (Jayasena et al., 2010).
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World cheese consumption has greatly increased during the last decade; there has been a
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shortage of milk production. Thus, the substitution and/or supplementation of cow milk with
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lupin milk would improve the yield and nutritional quality of cheese at a comparatively lower
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cost. Up to now, no work has been published on producing cheese using cow milk supplemented
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with lupin milk. Therefore, the aim of the present study was to investigate the effect of
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incorporation of lupin milk at different levels to cow milk on physicochemical, microbiological
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and sensory characteristics of cheese.
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2. Materials and methods
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2.1 Materials
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Seeds lupin (L. albus L.) of a local variety (Rubatab) were purchased from a local market
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(Omdurman, Sudan). Fresh cow milk samples were obtained from the University of Khartoum
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Farm, Shambat, Sudan. Rennet tablets were obtained from Christen Hansen’s Laboratorium
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(Copenhagen, Denmark). Unless otherwise stated all chemicals used in this study were of
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analytical grade.
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2.2 Preparation of lupin milk
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Lupin milk was extracted from the seeds by the method illustrated in Figure 1. Briefly,
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lupin seeds were cleaned manually to remove the dust, broken and infected seeds and then boiled
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in distilled water for 30 min to prevent seedling formation during soaking. The seed coat was
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removed by hand and then cotyledons were soaked in distilled water at a 1:3 (Cotyledons: water)
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ratio for 3 days at room temperature (25 ºC) with frequent changes of soaking water to washout
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alkaloids to decrease the bitter taste of the final product. At the end of the soaking period, the
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soaking water was decanted and the beans were washed. The hydrated cotyledons were then
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ground using a blender (Moulinex, Ecully, France) for 5 min at high speed to get lupin paste.
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Warm water (40-50 ºC) was added gradually during grinding until the total soluble solids of the
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extract reached 10 g/100 mL (0.5 kg/5 L water). The mixture was homogenized using ultrasonic
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homogenizer (Bandelin electronic GmbH and Co. KG, Berlin, Germany) and then centrifuged at
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2600 × g for 5 min. The extract was filtered through cheese cloth and then through 0.5 mm sieve.
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The resulting precipitate was dried in oven and then powdered using electric grinder to get lupin
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flour, whereas, the homogenized filtrate was considered as lupin milk. The milk was thermally
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treated at 100 ºC for 20 min to inactivate the lipoxygenase, and then cooled to 4 ºC. The milk
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stored at 4 ºC for further use.
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2.3 Cheese making process
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The cheese was prepared from pure cow milk or substituting cow milk at 25, 50, and 75
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mL/100 mL for lupin milk. The milk and the mixture (5 kg) were heated to 62 ºC for 30 min and
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then cooled to 40 ºC. Salt of a concentration of 6 mg/100g and rennet tablets at a rate of one
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tablet per 50 kg of milk were added. The mixture was stirred for 10 min and left for coagulation
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for 3 to 4 hrs. The coagulated curd was cut for whey separation, and then the curd of the cheese
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was poured into a wooden mould (50 x 50 x 20 cm) lined with a clean cloth and pressed by a
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heavy weight (5 kg) overnight. Then the cheese was removed from the mould and cut into small
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cubes. The whey of each cheese was collected in a separate container, boiled for five minutes,
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cooled and used for preservation of the cheese. To obtain a mature cheese, the cheese cubes
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were stored in sealed polyethylene bags at 5 ºC for 75 days. Both fresh and mature cheeses were
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analyzed for physicochemical and sensory characteristics.
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2.4 Chemical composition
The chemical composition of lupin seeds was determined according to the Standard
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Official Methods of Analysis (AOAC, 1990). Total carbohydrate of the seeds was calculated by
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difference. Fresh cow milk and lupin milk were analyzed for moisture, protein (Nx6.38 for cow’s
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milk and Nx5.8 for lupin’s milk) and ash following the standard methods (AOAC 1990). Protein
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and ash contents of cheese were also determined according to AOAC (1990) methods. Fat
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content of cow and lupin milk as well as that of cheeses was determined using Gerber tube
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following the method of Bradly, Arnold, Barbano, Semerad, Smith, & Vines (1992).
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2.5 Minerals determination
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Minerals were determined from the samples by the dry ashing method that is described by
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Chapman & Pratt (1982). Briefly, lupin seeds were dried and ashed at 500 °C under gradual
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increase in temperature (50 °C/h). About 2.0 g of samples was acid-digested with diacid mixture
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(HNO3 ⁄HClO4, 5:1, v ⁄ v) in a digestion chamber. The digested samples were dissolved in
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double-distilled water and filtered (Whatman No. 42). The filtrate was made to 50 mL with
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double-distilled water and was used for determination of total minerals. Sodium and potassium
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were determined according to the standard official methods (AOAC, 1990) using a Flame
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Photometer (Coring 410, Corning Medical and Scientific, Halstead Essex, UK). Calcium and
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phosphorous were determined following the method of Chapman and Pratt (1982).
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2.6 Determination of titratable acidity and pH
The acidity of cow and lupin milk before and after processing was determined according to
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the standard official methods (AOAC, 1990). About 10 ml of each were pipette into clean
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porcelain dish and five drops of phenolphthalein indicator were added and titrated with NaOH
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(0.1 mol equi/L). For the titratable acidity of cheese, 10 g of each type were weighed and placed
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in a conical flask. Distilled water at 40 ºC was added to the sample until the volume in the flask
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rose up to 105 mL. The flask was vigorously agitated and filtered through Whatman No.43 filter
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paper. Twenty five mL of the filtrate were pipetted in a 75 mL beaker and then five drops of
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phenolphthalein indicator were added to the filtrate and titrated against NaOH (0.1 mol equi/L)
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till a faint pink colour that lasted for about 30 seconds was obtained. The titratable acidity was
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then calculated as described in the official method (AOAC, 1990). The pH was determined using
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a pH meter, model pH 210 Microprocessor (Hanna instruments, Padova, Italy).
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2.7 Determination of total solids content
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Total solids content of the samples was determined according to AOAC (1990) methods.
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About 5 mL of each sample and 2 grams of cheese were weighed into three pairs of pre-weighed
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aluminum dishes. The weight of each sample and the dish was recorded. The dishes were put in
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an air oven at 100 ºC for 3 h, then placed in a desiccator to cool for 30 min and weighed. 8
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Heating, cooling and weighing were repeated several times to get a final weight of less than 0.5
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mg. Total solids content of each of the three samples was calculated as follows:
Total solids (g/100 g) =
Weight of sample after drying (g) × 100 Weight of sample before drying (g)
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2.8 Microbiological analysis
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All media were obtained in a dehydrated form and stored in a hygroscopic environment in a
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cool dry place away from light and prepared according to the manufacturer's instructions. The
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bacterial count was determined according to Houghtby, Maturin, and Koenig (1992) using
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standard plate count nutrient agar media. The plates were incubated at 37 ºC for 48 h. Typical
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colonies in the selected dilution were counted (25-250 colonies in each dilution). MacConkey
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broth purple medium was used for screening of coliform bacteria in both milk and cheese
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samples. The Salmonella was cultivated on nutrient broth media. The Wart agar media that
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contained chloramphenicol (200 mg/L) was used to screen moulds. For the determination of
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yeast and fungi MEA media contained chloramphenicol (200 mg/L) was used. The plates were
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incubated at 30 ºC for 48 h.
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2.9 Sensory evaluation
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The sensory characteristics of the cheeses were evaluated following the IDF standards
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(FIL-IDF 99B 1995). A trained panel of 12 members, composed of adult male (4; age ranged
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from 25 to 35) and female (8, age ranged from 26 to 40), was assigned to determine the quality 9
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of the fresh and mature cheese (colour, flavor, taste, texture and overall acceptability). Members
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were asked to score 1–5 hedonic scale (1 = poor, 2 = acceptable, 3= good, 4 = very good and 5 =
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excellent). The samples were randomized and presented using tag for each one. To determine the
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differences in judges’ response, the mean scores were analyzed by Duncan’s multiple range tests.
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2.10 Statistical analysis
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Statistical analyses were carried out using Minitab programme (1998). Three separate
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samples were analyzed and mean values were calculated. The data were assessed by analysis of
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variance (ANOVA) and by Duncan’s Multiple Range Test with a probability P ≤ 0.05.
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3. Results and discussion
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3.1 Biochemical composition of lupin seeds, lupin milk, cow milk and mixtures of cow and lupin
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milk
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The chemical composition of lupin seeds is shown in Table 1. The results showed that
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lupin seeds contain considerable amount of protein, carbohydrate, fat, and fiber. The contents of
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ash, moisture and macro-minerals such as Ca, Na, K, and P are comparable to those of many
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lupins. The protein content of lupin variety under investigation is very high compared to that
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reported for other lupin varieties (Jayasena et al., 2010). It has also been reported that seed
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protein content of wild populations of lupin varieties grown in Spain ranged from 23.8 to 33.6
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g/100 g (Pastor-Cavada, Juan, Pastor, Alaiz, & Vioque, 2009). Fat content of lupin seeds was
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10.4 g/100 g which is greater than that reported previously for some others lupin varieties
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(Vecerek, Suchy, Strakova & Machacek, 2008). Ash and fiber contents of local variety were
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lower than those reported for others lupins (Vecerek et al., 2008). Calcium and phosphorus were
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within the range of others lupins (Vecerek et al., 2008).
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Table 2 shows the results of the biochemical analysis of lupin milk, cow milk and the
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mixtures of cow and lupin milk. It is observed that lupin milk had higher (P ≤0.05) protein
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content than cow milk. As expected, the protein content of the mixture of cow and lupin milk is
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gradually increased (P ≤0.05) with increasing the concentration of lupin milk in the mixture. The
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difference in protein content of milk types is likely due to the fact that, during debittering
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process, carbohydrates, alkaloids, tannins and other compounds were partially washed out
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(Erbas, 2010; Jimenez-Martinez, Hernandez-Sanchez, Alvarez-Manilla, Robledo-Quintos,
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Martinez-Herrera, & Davila-Ortiz, 2001), thus causing an increase in the relative proportion of
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proteins. The protein content of lupin milk was 10 times lower than that of lupin seeds, but is
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still higher than that of cow milk. The decrease in protein content of lupin milk might be due to
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the removal of the seeds hull which may contain some proteins as well as the removal of soluble
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proteins during processing of the milk (Erbas, 2010). On the other hand, the fat content was
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higher in lupin milk than in cow milk, whereas, total solids are higher in cow milk compared to
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lupin milk. The fat content of the mixture of cow and lupin milk is significantly increased (P
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≤0.05) with increasing the concentration of lupin milk in the mixture. The total solids, on the
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other hand, was proportionally decreased (P ≤0.05) when the concentration of lupin milk in the
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mixture was increased. It is well known that, fat in milk helps to produce flavour, aroma and
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body in mature cheese (Horne & Banks, 2004). Both milk types showed non significant (P ≥
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0.05) difference with respect to acidity and pH. The reduction in the total solids of lupin milk
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might be due to the removal of the dry matter during milk processing as it has been previously
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reported that debittering reduced the total solids of lupin milk (Erbas, 2010; Jimenez-Martinez et
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al., 2001). The chemical composition of milk, especially the contents of protein, fat, calcium and
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pH, has a major influence on several aspects of cheese manufacture, especially rennet
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coagulability, gel strength, curd synergistic, and hence cheese composition and cheese yield
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(Fox, & Cogan, 2004).
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3.2 Lupin-based cheese formation and yield
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Substituting cow milk at 25, 50, and 75 mL/ 100 mL for lupin milk was observed to form
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cheese, without major effect on the curd formation compared to control cheese of pure cow milk
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(Fig. 2). Moreover, a remarkable increase in cheese yield was observed with increasing
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supplementation level of lupin milk. The mean value of cheese yield of pure cow milk was
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found to be 0.925 kg/ 5 kg milk and then gradually increased (P ≤0.05) to 0.975 kg/ 5 kg, 1.0
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kg/5 kg, and 1.12 kg/5kg with increasing lupin milk level to 25, 50 and 75 mL/100 mL,
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respectively. The enhancement of cheese yield could be attributed to the higher content of fat
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and protein in lupin milk compared to cow milk as well as low losses of total soluble solids in
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whey. In contrast, cheese yield decrease was reported in whey protein concentrate fortified milk
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cheese (Lobato-Calleros, Reyes-Hernandez, Beristain, Hornelas-Uribe, Sanchez-Garcia, &
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Vernon-Carter, 2007) and sesame protein isolate supplemented fresh cheese (Lu, Schmitt, &
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Chen, 2010). On the other hand, whey produced during cheese making was ranged from 2.5 to 3
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liter, with a lower level in cheese made from pure cow milk. This may likely be due to higher
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casein level in pure milk which leads to a slow rate of exclusion of whey or may be to Ca++
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activity which is an important factor in determining the rate of whey drainage. Additionally,
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slightly lower pH of lupin milk and the mixtures thereof could also contribute to the higher whey
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drainage of lupin based cheeses compared to the control cheese. Both higher casein
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concentration and pH were reported to decrease the curd syneresis and whey drainage (Daviau et
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al., 2000).
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3.3 Biochemical characteristics of lupin-based cheese
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Table 3 shows the biochemical characteristics of fresh soft cheese made with cow milk
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with and without lupin milk. The concentrations of protein, fat, total solids and acidity were
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significantly (P ≤0.05) increased with increasing lupin milk level in fresh soft cheese compared
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to pure milk fresh cheese. On the other hand, ash and pH showed significant (P ≤0.05) reduction
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as the level of lupin milk increased. Higher protein content (11.96 g/100 g) was observed when
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cow milk was mixed with 75 mL/100 mL lupin milk compared to that made from pure cow milk.
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The protein content of the soft cheese under investigation is slightly lower than that reported for
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Chinese traditional soy cheese known as sufu or furu (Ahmad, Yang, Ning, & Randhawa, 2008).
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As the level of lupin milk of fresh cheese increased over 25 mL/100 mL, fat content was
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significantly (P ≤0.05) increased which improved the quality of lupin containing cheese. Fats
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are known to play a major role in the rheology and texture as well as flavour of the product by
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acting as a source of fatty acids which in turn may be catabolized to flavour compounds, e.g.,
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methyl ketones, esters, thioesters and lactones (Collins, McSweeney, & Wilkinson, 2003). Total
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solids content of fresh cheese was significantly (P ≤0.05) increased with increase in the level of
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lupin milk. This might be due to lower water content of lupin-based cheese compared to that of
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control cheese as the previous result showed that the whey exclusion of lupin-based cheese was
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higher compared to that of the control cheese. The increase in total solids in lupin-based cheese
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could also be attributed to higher protein and fat contents in lupin milk compared to cow milk.
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The results obtained were within the range reported for white cheese prepared from cow milk (El
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Owni & Hamed, 2009). As the level of lupin milk increased in fresh cheese a significant (P
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≤0.05) reduction in ash content was observed compared to that of pure milk which could be
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attributed to the lower amount of ash in lupin milk. For unknown reasons, the acidity of fresh
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cheese was significantly (P ≤0.05) increased when the level of lupin milk was increased.
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Significantly (P ≤0.05) low acidity value was observed in soft fresh cheese made from pure cow
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milk, compared to that obtained for cheese containing lupin milk. This could be attributed to
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change in the proportion of lactose which is converted by lactic acid bacteria to lactic acid. The
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complete and rapid metabolism of lactose in cheese curd is essential for the production of good
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quality cheese since the presence of a fermentable carbohydrate may lead to the development of
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an undesirable secondary flora (Fox, Guinee, Cogan, & McSweeney, 2000). Since low acidity
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curd normally contains more moisture than high acidity curd, one might expect more syneresis
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and higher whey drainage of low acidity curd. The pH of fresh soft cheese was significant (P
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≤0.05) decreased when the level lupin milk was increased in the cheese compared to that of fresh
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cheese prepared from pure milk. The reduction of the pH could be attributed to the increased
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acidity of the cheese with higher level of lupin milk. Since the increase in the acidity is always
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contradictory with the pH.
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Table 4 shows the impact of maturation for 75 days on biochemical characteristics of pure
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milk cheese and that manufactured by addition of lupin milk of different levels. Mature cheese
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prepared from a mixture of pure milk and lupin milk contained higher (P ≤0.05) amount of
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protein compared to that prepared from pure milk. When compared to fresh soft cheese, a 14
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remarkable reduction in the protein content was observed in all types of mature cheese. Despite
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inactivation of the enzymes by heating, the reduction in protein content could be due to
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hydrolysis of the protein in cheese by the residual chymosin, indigenous proteinases and/or
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microbial proteinases from starter and non-starter microorganisms. Primary proteolysis of cheese
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proteins is mainly the result of the action of endogenous proteinases (i.e., plasmin and somatic
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cell proteinases) and residual coagulant. However, proteinases from starter and nonstarter
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microorganisms can also be active in the degradation of cheese proteins and peptides. Lactic acid
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bacteria are weakly proteolytic, but possess a very comprehensive range of proteinases and/or
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peptidases capable of hydrolyzing casein-derived peptides to small peptides and amino acids.
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Proteolysis is the most complex, and in most cases, the most important biochemical event which
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occurs during cheese ripening and it contributes to the development of cheese texture as well as
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flavour (Sousa, Ardo, & McSweeney, 2001). Fat content of mature control cheese as well as that
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contain 25 mL/100 mL lupin milk was slightly increased compared to fresh soft cheese.
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However, the fat content of mature cheese significantly (P ≤0.05) decreased with increase in
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lupin milk level compared to that of cheese prepared from pure milk. Ash content of mature
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cheese significantly (P ≤0.05) increased at all levels of lupin milk compared to fresh cheese. The
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increases in the ash content during storage might be due to the decrease in moisture content as
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well as absorption of salt by cheese curd. Total solids increased in mature cow milk cheese
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compared to the fresh one. However, mature cheese prepared from a mixture of lupin and cow
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milk had low total solids compared to fresh cheese. Moreover, as the level of lupin milk
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increased, total solids of mature cheese were decreased. The acidity of mature cheese prepared
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from pure milk was significantly (P ≤0.05) increased compared to fresh cheese. The
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enhancement of the acidity of pure milk cheese might be due to the formation of lactic acid by
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lactic acid bacteria during fermentation of lactose. However, addition of lupin milk significantly
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(P ≤0.05) reduced the acidity of mature cheese compared to fresh one. This might be due to the
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lack and/or low concentration of fermentable sugar (lactose) in lupin containing cheeses. The
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complete and rapid metabolism of the lactose and its constituent monosaccharides in cheese curd
324
is essential for the production of good quality cheese since the presence of a fermentable
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carbohydrate may lead to the development of an undesirable secondary flora during ripening.
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The acidity depends primarily on type and quantity of starter added, pre-acidification applied,
327
temperature and duration of acid development. The pH of lupin containing mature cheese was
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significantly (P ≤0.05) higher than that of fresh one. As the level of lupin milk increased the pH
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of mature cheese gradually (P ≤0.05) decreased and reached 6.10 at 75 mL/100 mL level but still
330
higher than that of fresh cheese. The rises of the pH of cheeses during ripening could be
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attributed to the formation of alkaline nitrogenous compounds (such as ammonia) and/or
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catabolism of lactic acid and thus reduce the acidity of the cheeses.
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3.4 Microbial analysis lupin-based cheese
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Table 5 shows the microbial evaluation of cheeses made from cow milk mixed with
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different levels of lupin milk. The total bacterial count is within the range that naturally exists in
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milk containing foods. Compared to pure milk cheese, cheese made with 75 mL/100 mL lupin
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milk had lower bacterial count followed by that made of 25 mL/100 mL lupin milk. Higher
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bacterial count was observed when cow milk substituted with 50 mL/100 mL for lupin milk.
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Other microorganisms such as fungi, mould, E. coli, and Salmonella were not detected. Similar
341
results of bacterial count as well as absence of fungi, mould, E. coli and salmonella were also
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observed in lupin milk before incorporation into cheeses (data not shown). The absence of such
343
microorganisms in both lupin milk and the cheese made thereof may be due to the extraction
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method used for the preparation of lupin milk in which clean water was used for the extraction as
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well as pasteurization of the milk used in the cheese making. Similar observation was observed
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in vegetable-based dairy products (Aminigo, Metzger, & Lehtola, 2009). In addition to the use of
347
high temperature, high level of NaCl (1.4–2.6 mol/L) when added to the cheese milk will control
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the growth of the indigenous microflora (Mohamed Ahmed et al., 2010).
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3.5 Sensory characteristics of lupin-based cheese
The sensory characteristics of fresh soft cheese made using different levels of lupin milk
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are shown in Table 6. The cheese prepared by adding different levels of lupin milk was not
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significantly (P ≥ 0.05) differed from that of pure milk with respect to taste and texture except
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that prepared from 75 mL/100 mL lupin milk which had lower scores of taste and texture. Higher
355
taste score was obtained for cheese made with 25 mL/100 mL lupin milk and the lower one was
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observed for cheese made with 75mL/100 mL lupin milk. Moreover, no significant (P ≥ 0.05)
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differences were observed in the color of all types of cheese. Addition of lupin milk at the level
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of 50 and 75 mL/100 mL resulted in low score of the color of the cheese compared to addition of
359
25 mL/100 mL lupin milk and that of pure milk. Incorporation of lupin milk in cheese
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significantly (P ≤0.05) improved the flavour of it. Higher score of flavor was obtained for
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cheese made with 75 mL/100 mL lupin milk whereas the lower one was observed in cheese
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prepared from pure milk. This probably could be due to the degradation of carbohydrates to
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lactic acid and release of flavour components such as acetaldehyde or decomposition of fat into
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volatile fatty acid. There are significant (P ≤0.05) differences in the overall acceptability
365
between pure milk cheese and those prepared by adding 50 and 75 mL/100 mL lupin milk. Panel
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test showed that fresh cheese prepared from 50 and 75 mL/100 mL lupin milk are less acceptable
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compared to that of pure milk and that prepared by mixing 25 mL/100 mL lupin milk. Strikingly,
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incorporation of lupin milk in the cheese at low concentration (25 mL/100 mL) does not caused
369
any defects, and even slightly improved, the sensory characteristics of fresh soft cheese
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compared to the control cheese made of pure cow milk. Whereas, high concentration (75 mL/
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100 mL) of lupin milk significantly (P ≤0.05) affect the sensory characteristics of fresh soft
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cheese compared to the control cheese and that made of 25 mL/100 mL lupin milk.
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Table 7 shows the sensory characteristics of mature cheese prepared from cow milk with or
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without lupin milk. It is noted that some sensory characteristics such as taste, texture and overall
375
acceptability in cheese made with 25 mL/100 mL lupin milk were improved during maturation
376
except colour and flavour compared to fresh cheese. For mature cheese of pure milk the taste,
377
color and overall acceptability were improved with exception of flavor and texture. Generally,
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the overall acceptability of all types of cheese was enhanced during maturation. The results
379
obtained agree with the findings of Tarakci and Kucukoner (2006) who reported that the overall
380
acceptability of cheese was increased during ripening. The results also showed that there are
381
significant (P ≤0.05) differences in the scores of taste, texture, flavor and overall acceptability
382
between all types of cheese including the control one. In lupin containing cheese, the increase of
383
lupin milk concentration significantly (P ≤0.05) reduced the scores of taste, texture and overall
384
acceptability of the cheese but it significantly (P ≤0.05) improved the flavor. Lowest scores of
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taste, texture, color, and overall acceptability among all cheeses including the control one were
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observed in cheese made with 75 mL/100 mL of lupin milk. Interestingly, incorporation of lupin
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milk in the cheese at low concentration (25 mL/100 mL) significantly (P ≤0.05) enhanced the
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taste, texture, flavor, and overall acceptability of the final mature cheese compared to the control
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cheese. With an exception of the flavor, at higher concentrations (50 and 75 mL/100 mL) of
390
lupin milk in the cheese these all sensory attributes were significantly (P ≤0.05) decreased
391
compare to those of control cheese. Overall, substitution of cow milk at 25 mL/100 mL for lupin
392
milk significantly enhanced most of the sensory attributes of final mature cheese, with slight
393
defect in the color compared to the control cheese of pure cow milk. The sensory characteristics
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of a cheese at the time of its consumption reflect the quality of milk from which it was produced,
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the processes used for the production and the physical and chemical changes that may occurred
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during maturation (Delahunty & Drake, 2004).
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4. Conclusion
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The results of this study demonstrated that lupin milk containing cheese at low substitution
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of cow milk (25 mL/100 mL) for lupin milk has close resemblance to that of cow milk cheese in
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some nutritional and sensory characteristics. Substitution of cow milk at 25 mL/100 mL for lupin
402
milk can be used for the production of white soft cheese without causing any major changes in
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quality attributes of both fresh and mature cheese. Moreover, incorporation of lupin milk in
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cheese manufactured resulted in cost saving and improvement of the nutritional value.
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References
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Johnson, S.K., Chua, V., Hall R.S., & Baxter A.L. (2006). Lupin kernel fibre foods improve
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bowel function and beneficially modify some putative faecal risk factors for colon cancer in
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Lee, Y.P., Mori, T., Sipsas, S., Barden, A., Puddey, I., Burke, V., Hall R., & Hodgson, J. (2006).
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J.E., & Vernon-Carter, E.J. (2007). Microstructure and texture of white fresh cheese made
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Lu, X., Schmitt, D., & Chen, S. (2010). Effect of sesame protein isolate in partial replacement of
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Martins, J.M., Riottot, M., de Abreu, M.C., Viegas-Crespo, A.M., Lança, M.J., Almeida, J.A.,
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Freire, J.B., & Bento, O.P. (2005). Cholesterol-lowering effects of dietary blue lupin
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Pastor-Cavada, E., Juan, R., Pastor, J. E., Alaiz, M., & Vioque, J. (2009). Analytical nutritional
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characteristics of seed proteins in six wild Lupinus species from Southern Spain. Food
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during cheese ripening. International Dairy Journal, 11, 327–345. Vecerek, V., Suchy, P., Strakova, E. & Machacek, M. (2008). Nutritive composition of seeds of
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the lupin varieties registered in the Czech republic. In: J.A. Palta and J.B. Berger (eds).
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Tarakci, Z., & Kucukoner, E. (2006). Changes in physicochemical, lipolysis and proteolysis of
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vacuum packed Turkish Kashar cheese during ripening. Journal of Central European
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Agriculture, 7, 459-464.
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Table 1. Chemical composition of the local variety of lupin seeds (dry weight) and
509
other lupines.
(g/100 g)
Local variety
Other Lupins*
(Rubatab) 46.3(±2.31)
Fat
10.4(±1.14)
Carbohydrate
27.1(±1.26)
Ash
1.6(±0.10)
Moisture
4.6(±0.11)
Fiber Mineral composition (g/100 g) Ca Na
P
4.5-8.0 ND
3.77-5.2 ND
10.0(±1.15)
12.8-16.9
0.4(±0.03)
0.27-0.48
0.01(±0.02)
ND
0.85(±0.1)
ND
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33.2-43.7
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Chemical composition
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0.86(±0.09)
*Vecerek et al. (2008). ND; not detected
511
Values are means of triplicate samples (±SD)
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0.55-0.92
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Parameter
Lupin milk
Cow milk
Protein (g/100 g)
4.90 (±0.10)
Fat (g/100 g)
Concentration of lupin milk (mL/100 mL) 50
75
3.90 (±0.08)
4.10 (±0.06)
4.50 (±0.09)
4.70 (±0.10)
5.00 (±0.12)
3.80 (±0.13)
4.20 (±0.08)
4.40 (±0.08)
4.80 (±0.12)
Acidity (% lactic acid)
0.30 (±0.01)
0.20 (±0.01)
0.23 (±0.04)
0.25 (±0.05)
0.26 (±0.05)
Total solids (g/100 g)
11.20 (±0.07)
12.60(±0.11)
12.30 (±0.18)
12.0 (±0.13)
11.40 (±0.10)
pH
6.30 (±0.10)
6.40 (±0.06)
6.38 (±0.05)
6.35 (±0.06)
6.31(±0.02)
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Values are means of triplicate samples (±SD)
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Table 2. Biochemical composition of lupin milk, cow milk and mixtures of cow and lupin milk
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Table 3. Biochemical composition (g/100 g) of fresh soft cheese made from cow milk supplemented with lupin milk of different levels. Concentration of lupin milk (mL/100 mL) 25
50
Protein
6.66c (±0.07)
8.90b(±0.12)
8.96b(±0.10)
11.96a(±0.11)
Fat
10.00c(±0.10)
10.00c(±0.11)
12.00b(±0.15)
12.33a(±0.18)
Ash
2.49a(±0.05)
1.13b(±0.06)
0.89c(±0.01)
0.69d(±0.03)
Total solids
38.23c(±0.64)
40.40b(±0.69)
40.63b(±0.61)
42.30a(±0.76)
Acidity (% lactic acid)
0.18d(±0.01)
0.80c(±0.03)
1.66b(±0.04)
2.66a(±0.08)
pH
6.43a(±0.09)
5.16d(±0.08)
5.46b(±0.10)
5.23c(±0.06)
Values are means of triplicates (±SD). are significantly different at (P ≤0.05).
a-d
SC
:Means average in the same raw with different letters
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Parameter
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519 520
Table 4. Biochemical composition (g/100 g) of mature soft cheese made from cow milk supplemented with lupin milk of different levels. Concentration of lupin milk (mL/100 mL) 25
50
Protein
3.03d(±0.04)
4.23b(±0.05)
4.70a(±0.07)
4.11c(±0.04)
Fat
11.66a(±0.13)
11.33b(±0.09)
10.33d(±0.11)
10.66c(±0.07)
Ash
3.19a(±0.06)
1.09d(±0.01)
1.46c(±0.03)
1.56b(±0.04)
Total solids
39.80a(±0.55)
39.20b(±0.23)
38.33c(±0.32)
37.80d(±0.19)
Acidity (% lactic acid)
0.80a(±0.03)
0.70b(±0.02)
0.40d(±0.01)
0.50c(±0.01)
pH
6.70a(±0.05)
6.63b(±0.04)
6.30c(±0.04)
6.10d(±0.03)
Values are means of triplicates (±SD). are significantly different at (P ≤0.05).
a-d
SC
:Means average in the same raw with different letters
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75
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Parameter
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Table 5. Microbial analysis of soft cheese made from cow milk supplemented with different levels of lupin milk Test
Bacterial count 526
Concentration of lupin milk (mL/100 mL) 0
25
50
3.3× 106 cfu/ml
2.6× 106 cfu/ml
5.6× 106 cfu/ml
Other microbes such as Fungi, Yeast, E. coli and Salmonella were not detected.
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30
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1.3× 106 cfu/ml
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528 529
Table 6. Sensory characteristics of fresh soft cheese made from cow milk supplemented with different levels of lupin milk Concentration of lupin milk (mL/100 mL) 25
50
75
Taste
3.50a(±0.04)
3.58a(±0.08)
3.50a(±0.05)
2.91b(±0.03)
Texture
3.50a(±0.03)
3.50a(±0.04)
3.58a(±0.09)
2.91b(±0.01)
Color
3.33a(±0.07)
3.33a(±0.09)
3.16a(±0.08)
3.16a(±0.10)
Flavor
2.75c(±0.02)
3.16b(±0.04)
3.16b(±0.06)
3.58a(±0.09)
Overall acceptability
3.00a(±0.03)
3.08a(±0.05)
2.66b(±0.01)
2.25c(±0.04)
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Values are means of 12 samples (±SD). a-c :Means average in the same raw with different letters are significantly different at (P ≤0.05). Hedonic scale (1–5, 1 = poor, 5 = excellent).
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Quality attribute
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Table 7. Sensory characteristics of mature soft cheese made from cow milk supplemented with different levels of lupin milk
0
25
50
Taste
3.75b(±0.02)
4.16a(±0.04)
3.08c(±0.02)
2.41d(±0.03)
Texture
3.50b(±0.03)
3.58a(±0.04)
3.25c(±0.01)
2.25d(±0.04)
Color
4.00a(±0.10)
3.25b(±0.07)
3.33b(±0.06)
3.16b(±0.08)
Flavor
2.75d(±0.01)
3.08c(±0.05)
3.58b(±0.05)
3.83a(±0.02)
Overall acceptability
3.75b(±0.03)
4.16a(±0.09)
3.08c(±0.04)
2.41d(±0.01)
544
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543
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541
75
Values are means of 12 samples (±SD). a-d :Means average in the same raw with different letters are significantly different at (P ≤0.05). Hedonic scale (1–5, 1 = poor, 5 = excellent).
538
540
SC
Quality attribute
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Concentration of lupin milk (mL/100 mL)
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Fig.1. Preparation of lupin milk from the seeds of Lupinus albus
547 548 549 33
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Fig. 2. White soft cheese produced using cow milk supplemented with lupin milk at different concentrations of 0 mL/100 mL (A), 25 mL/100 mL (B), 50 mL/100 mL (C), and 75 mL/100 mL (D).
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Physicochemical, microbiological and sensorial properties soft cheese made of mixture of cow and lupin milk were evaluated.
Lupin milk based cheese at low substitution (25 mL/100 mL) for lupin milk
sensory characteristics.
Substitution of cow milk at 25 mL/100 mL for lupin milk could be used without major changes in the quality of final cheese.
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has close resemblance to that of cow milk cheese in terms of biochemical and
Incorporating lupin milk into cheese could save money as well as improve the
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nutritional value of such product.
ACCEPTED MANUSCRIPT Research Highlight
Quality attributes of cheese made of mixture of cow and lupin milk were evaluated.
Substitution of cow milk at 25 mL/100 mL for lupin milk produced good quality cheese.
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Incorporating lupin milk into cheese could save money.
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S0023-6438(14)00232-1
DOI:
10.1016/j.lwt.2014.04.027
Reference:
YFSTL 3863
To appear in:
LWT - Food Science and Technology
Received Date: 10 November 2011 Revised Date:
21 July 2012
Accepted Date: 13 April 2014
Please cite this article as: Elsamani, M.O., Habbani, S.S., Babiker, E.E., Mohamed Ahmed, I.A., Biochemical, microbial and sensory evaluation of white soft cheese made from mixture cow and lupin milk, LWT - Food Science and Technology (2014), doi: 10.1016/j.lwt.2014.04.027. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
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Biochemical, microbial and sensory evaluation of white
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soft cheese made from cow and lupin milk
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Mohamed O. Elsamania, Siddig S. Habbania, Elfadil E. Babikerb and Isam A. Mohamed
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Ahmedc,d* a
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Department of Food Science and Technology, Faculty of Agriculture, Omdurman Islamic University, Omdurman, Sudan
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Department of Food Science and Nutrition, College of Food and Agricultural Sciences, King Saud University, P. O. Box 2460, Riyadh 11451, Kingdom of Saudi Arabia
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Department of Food Science and Technology, Faculty of Agriculture, University of Khartoum, Khartoum North, Shambat
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Department of Applied Resources Chemistry, Faculty of Agriculture, Tottori University, Tottori 680-8553, Japan
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E-mail: [email protected]
Corresponding author: Tel.:+81 857 31 5443; Fax: +81 857 31 5347
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Abstract
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The aim of the present study was to evaluate some physicochemical, microbiological and sensory
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properties of fresh and matured (75 days) soft cheeses made with mixtures of cow milk and 0,
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25, 50 and 75 mL/100 mL of lupin milk. A remarkable increase in cheese yield was observed
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with increasing the level of lupin milk to the mixture. Compared to cow milk cheese, the protein
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content was significantly (P ≤ 0.05) increased while ash was decreased with the increase in the
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level of lupin milk for both fresh and matured cheese. However, fat content, total solids and
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acidity were increased only for fresh cheese and decreased for mature one compared to that of
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cow milk. The pH showed significant (P ≤ 0.05) reduction when the levels of lupin milk
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increased for fresh cheese while for matured cheese it slightly decreased. The total bacterial
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count is within the range that naturally exists in milk containing foods. The others
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microorganisms such as fungi, mould, E. coli, and Salmonella were not existed in both types of
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cheese. Regardless of cheese colour, incorporation of lupin milk at low concentration (25
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mL/100 mL) significantly (P ≤ 0.05) enhanced the taste, texture, flavor, and overall acceptability
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of both fresh and mature cheese.
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Keyword: Legumes, Lupinus albus, Lupin milk, sensory quality, white soft cheese
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1. Introduction Species of the genus Lupinus have been used for human nutrition for over 6000 years. Of
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them, white lupin (Lupinus albus L.) is one of the oldest agricultural crops widely used in the
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world not only as a protein source in fodder production but also for soil improvement (Huyghe,
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1997). In the past decades, the production and consumption of white lupin have increased due to
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the capability of lupin plants to grow in adverse climates, their tolerance to poor soils and their
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high crop yields (1000–2000 kg/ha) (Linnemann & Dijkstra, 2002). The protein content of lupin
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seeds ranges from 30 to 40 g/100 g with an amino acid composition comparable to that of soy
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bean. Although lupin seeds are mainly used as a feed, they are acquiring increasing importance
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as a source of vegetable proteins for human consumption in many countries. Hence, increased
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consumer awareness on health issues has produced a dramatic and fast-growing demand of plant
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protein food products. In particular, lupin proteins are receiving attention in terms of health
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benefits, particularly in relation to a number of conditions now known as ‘metabolic syndrome’
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which includes a cluster of factors such as, obesity, high blood pressure, insulin resistance and
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elevated blood cholesterol (Arnoldi, 2005). Lupin enriched foods have the potential to:
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beneficially influence satiety (appetite suppression) and energy balance (Lee et al., 2006),
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beneficially influence glycaemic control (Hall, Thomas & Johnson, 2005), improve blood lipids
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(Martins et al., 2005), reduce hypertension (Pilvi, Jauhiainen, Cheng, Mervaala, Vapaatalo, &
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Korpela, 2006) and improve bowel health (Johnson, Chua, Hall & Baxter, 2006). Today, a
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market for the use of lupin seeds in food has been developed and different products are available,
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such as pasta, bakery products containing lupin flour, meat products, beverages and tofu
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(Jayasena, Khu, & Nasar-Abbas, 2010; Paraskevopoulou, Provatidou, Tsotsiou, & Kiosseoglou,
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2010).
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Cheese is a dairy product that has played a key role in human nutrition for centuries. The
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broad range of different cheeses available is based mainly on regional conditions and production
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technology, which has been repeatedly adapted and optimized. The main objective has always
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been and still is to convert milk, which is perishable, into a product with a longer shelf life whilst
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preserving most of its nutrients (Mohamed Ahmed, Babiker, & Mori, 2010). The growth in the
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ready meals sector in recent years had been reflected in the increase in the demand for cheese as
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a food ingredient. Shredded, diced, sliced, and even liquid cheese has been developed to meet the
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needs of the modern food industry, as convenience foods continue to grow in popularity (Byrne,
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1999). The need to increase the amount of cheese from a given amount of milk is an economic
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necessity when the milk supply is limited, but the demand for such product is high. Research
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efforts have been directed mainly toward evaluating procedures for using soy bean protein
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isolate as an extender in cheeses with limited success due to either the difficulty in incorporating
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the soy protein isolate into the finished cheese or because of quality issues with the finished
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product. Seeds of other plants (such as Lupinus, reported as a possible substitute of soy in human
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foodstuffs) have not received enough attention, not because they do not contain proteins that may
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be functionally or nutritionally equal to soy bean, but rather because of economic reasons.
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However, this situation may change, and it seems advisable at present to explore the
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development and utilization of a variety of foods derived from or containing a wide range of
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plant proteins. Recent attempts have been made to process lupin seeds to produce milk-like,
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dairy replacement products, similar to those derived from soybeans (Jayasena et al., 2010).
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World cheese consumption has greatly increased during the last decade; there has been a
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shortage of milk production. Thus, the substitution and/or supplementation of cow milk with
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lupin milk would improve the yield and nutritional quality of cheese at a comparatively lower
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cost. Up to now, no work has been published on producing cheese using cow milk supplemented
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with lupin milk. Therefore, the aim of the present study was to investigate the effect of
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incorporation of lupin milk at different levels to cow milk on physicochemical, microbiological
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and sensory characteristics of cheese.
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2. Materials and methods
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2.1 Materials
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Seeds lupin (L. albus L.) of a local variety (Rubatab) were purchased from a local market
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(Omdurman, Sudan). Fresh cow milk samples were obtained from the University of Khartoum
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Farm, Shambat, Sudan. Rennet tablets were obtained from Christen Hansen’s Laboratorium
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(Copenhagen, Denmark). Unless otherwise stated all chemicals used in this study were of
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analytical grade.
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2.2 Preparation of lupin milk
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Lupin milk was extracted from the seeds by the method illustrated in Figure 1. Briefly,
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lupin seeds were cleaned manually to remove the dust, broken and infected seeds and then boiled
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in distilled water for 30 min to prevent seedling formation during soaking. The seed coat was
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removed by hand and then cotyledons were soaked in distilled water at a 1:3 (Cotyledons: water)
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ratio for 3 days at room temperature (25 ºC) with frequent changes of soaking water to washout
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alkaloids to decrease the bitter taste of the final product. At the end of the soaking period, the
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soaking water was decanted and the beans were washed. The hydrated cotyledons were then
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ground using a blender (Moulinex, Ecully, France) for 5 min at high speed to get lupin paste.
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Warm water (40-50 ºC) was added gradually during grinding until the total soluble solids of the
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extract reached 10 g/100 mL (0.5 kg/5 L water). The mixture was homogenized using ultrasonic
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homogenizer (Bandelin electronic GmbH and Co. KG, Berlin, Germany) and then centrifuged at
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2600 × g for 5 min. The extract was filtered through cheese cloth and then through 0.5 mm sieve.
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The resulting precipitate was dried in oven and then powdered using electric grinder to get lupin
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flour, whereas, the homogenized filtrate was considered as lupin milk. The milk was thermally
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treated at 100 ºC for 20 min to inactivate the lipoxygenase, and then cooled to 4 ºC. The milk
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stored at 4 ºC for further use.
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2.3 Cheese making process
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The cheese was prepared from pure cow milk or substituting cow milk at 25, 50, and 75
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mL/100 mL for lupin milk. The milk and the mixture (5 kg) were heated to 62 ºC for 30 min and
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then cooled to 40 ºC. Salt of a concentration of 6 mg/100g and rennet tablets at a rate of one
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tablet per 50 kg of milk were added. The mixture was stirred for 10 min and left for coagulation
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for 3 to 4 hrs. The coagulated curd was cut for whey separation, and then the curd of the cheese
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was poured into a wooden mould (50 x 50 x 20 cm) lined with a clean cloth and pressed by a
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heavy weight (5 kg) overnight. Then the cheese was removed from the mould and cut into small
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cubes. The whey of each cheese was collected in a separate container, boiled for five minutes,
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cooled and used for preservation of the cheese. To obtain a mature cheese, the cheese cubes
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were stored in sealed polyethylene bags at 5 ºC for 75 days. Both fresh and mature cheeses were
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analyzed for physicochemical and sensory characteristics.
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2.4 Chemical composition
The chemical composition of lupin seeds was determined according to the Standard
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Official Methods of Analysis (AOAC, 1990). Total carbohydrate of the seeds was calculated by
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difference. Fresh cow milk and lupin milk were analyzed for moisture, protein (Nx6.38 for cow’s
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milk and Nx5.8 for lupin’s milk) and ash following the standard methods (AOAC 1990). Protein
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and ash contents of cheese were also determined according to AOAC (1990) methods. Fat
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content of cow and lupin milk as well as that of cheeses was determined using Gerber tube
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following the method of Bradly, Arnold, Barbano, Semerad, Smith, & Vines (1992).
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2.5 Minerals determination
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Minerals were determined from the samples by the dry ashing method that is described by
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Chapman & Pratt (1982). Briefly, lupin seeds were dried and ashed at 500 °C under gradual
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increase in temperature (50 °C/h). About 2.0 g of samples was acid-digested with diacid mixture
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(HNO3 ⁄HClO4, 5:1, v ⁄ v) in a digestion chamber. The digested samples were dissolved in
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double-distilled water and filtered (Whatman No. 42). The filtrate was made to 50 mL with
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double-distilled water and was used for determination of total minerals. Sodium and potassium
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were determined according to the standard official methods (AOAC, 1990) using a Flame
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Photometer (Coring 410, Corning Medical and Scientific, Halstead Essex, UK). Calcium and
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phosphorous were determined following the method of Chapman and Pratt (1982).
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2.6 Determination of titratable acidity and pH
The acidity of cow and lupin milk before and after processing was determined according to
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the standard official methods (AOAC, 1990). About 10 ml of each were pipette into clean
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porcelain dish and five drops of phenolphthalein indicator were added and titrated with NaOH
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(0.1 mol equi/L). For the titratable acidity of cheese, 10 g of each type were weighed and placed
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in a conical flask. Distilled water at 40 ºC was added to the sample until the volume in the flask
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rose up to 105 mL. The flask was vigorously agitated and filtered through Whatman No.43 filter
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paper. Twenty five mL of the filtrate were pipetted in a 75 mL beaker and then five drops of
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phenolphthalein indicator were added to the filtrate and titrated against NaOH (0.1 mol equi/L)
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till a faint pink colour that lasted for about 30 seconds was obtained. The titratable acidity was
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then calculated as described in the official method (AOAC, 1990). The pH was determined using
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a pH meter, model pH 210 Microprocessor (Hanna instruments, Padova, Italy).
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2.7 Determination of total solids content
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Total solids content of the samples was determined according to AOAC (1990) methods.
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About 5 mL of each sample and 2 grams of cheese were weighed into three pairs of pre-weighed
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aluminum dishes. The weight of each sample and the dish was recorded. The dishes were put in
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an air oven at 100 ºC for 3 h, then placed in a desiccator to cool for 30 min and weighed. 8
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Heating, cooling and weighing were repeated several times to get a final weight of less than 0.5
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mg. Total solids content of each of the three samples was calculated as follows:
Total solids (g/100 g) =
Weight of sample after drying (g) × 100 Weight of sample before drying (g)
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2.8 Microbiological analysis
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All media were obtained in a dehydrated form and stored in a hygroscopic environment in a
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cool dry place away from light and prepared according to the manufacturer's instructions. The
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bacterial count was determined according to Houghtby, Maturin, and Koenig (1992) using
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standard plate count nutrient agar media. The plates were incubated at 37 ºC for 48 h. Typical
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colonies in the selected dilution were counted (25-250 colonies in each dilution). MacConkey
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broth purple medium was used for screening of coliform bacteria in both milk and cheese
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samples. The Salmonella was cultivated on nutrient broth media. The Wart agar media that
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contained chloramphenicol (200 mg/L) was used to screen moulds. For the determination of
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yeast and fungi MEA media contained chloramphenicol (200 mg/L) was used. The plates were
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incubated at 30 ºC for 48 h.
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2.9 Sensory evaluation
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The sensory characteristics of the cheeses were evaluated following the IDF standards
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(FIL-IDF 99B 1995). A trained panel of 12 members, composed of adult male (4; age ranged
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from 25 to 35) and female (8, age ranged from 26 to 40), was assigned to determine the quality 9
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of the fresh and mature cheese (colour, flavor, taste, texture and overall acceptability). Members
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were asked to score 1–5 hedonic scale (1 = poor, 2 = acceptable, 3= good, 4 = very good and 5 =
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excellent). The samples were randomized and presented using tag for each one. To determine the
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differences in judges’ response, the mean scores were analyzed by Duncan’s multiple range tests.
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2.10 Statistical analysis
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Statistical analyses were carried out using Minitab programme (1998). Three separate
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samples were analyzed and mean values were calculated. The data were assessed by analysis of
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variance (ANOVA) and by Duncan’s Multiple Range Test with a probability P ≤ 0.05.
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3. Results and discussion
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3.1 Biochemical composition of lupin seeds, lupin milk, cow milk and mixtures of cow and lupin
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milk
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The chemical composition of lupin seeds is shown in Table 1. The results showed that
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lupin seeds contain considerable amount of protein, carbohydrate, fat, and fiber. The contents of
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ash, moisture and macro-minerals such as Ca, Na, K, and P are comparable to those of many
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lupins. The protein content of lupin variety under investigation is very high compared to that
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reported for other lupin varieties (Jayasena et al., 2010). It has also been reported that seed
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protein content of wild populations of lupin varieties grown in Spain ranged from 23.8 to 33.6
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g/100 g (Pastor-Cavada, Juan, Pastor, Alaiz, & Vioque, 2009). Fat content of lupin seeds was
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10.4 g/100 g which is greater than that reported previously for some others lupin varieties
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(Vecerek, Suchy, Strakova & Machacek, 2008). Ash and fiber contents of local variety were
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lower than those reported for others lupins (Vecerek et al., 2008). Calcium and phosphorus were
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within the range of others lupins (Vecerek et al., 2008).
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Table 2 shows the results of the biochemical analysis of lupin milk, cow milk and the
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mixtures of cow and lupin milk. It is observed that lupin milk had higher (P ≤0.05) protein
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content than cow milk. As expected, the protein content of the mixture of cow and lupin milk is
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gradually increased (P ≤0.05) with increasing the concentration of lupin milk in the mixture. The
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difference in protein content of milk types is likely due to the fact that, during debittering
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process, carbohydrates, alkaloids, tannins and other compounds were partially washed out
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(Erbas, 2010; Jimenez-Martinez, Hernandez-Sanchez, Alvarez-Manilla, Robledo-Quintos,
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Martinez-Herrera, & Davila-Ortiz, 2001), thus causing an increase in the relative proportion of
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proteins. The protein content of lupin milk was 10 times lower than that of lupin seeds, but is
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still higher than that of cow milk. The decrease in protein content of lupin milk might be due to
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the removal of the seeds hull which may contain some proteins as well as the removal of soluble
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proteins during processing of the milk (Erbas, 2010). On the other hand, the fat content was
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higher in lupin milk than in cow milk, whereas, total solids are higher in cow milk compared to
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lupin milk. The fat content of the mixture of cow and lupin milk is significantly increased (P
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≤0.05) with increasing the concentration of lupin milk in the mixture. The total solids, on the
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other hand, was proportionally decreased (P ≤0.05) when the concentration of lupin milk in the
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mixture was increased. It is well known that, fat in milk helps to produce flavour, aroma and
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body in mature cheese (Horne & Banks, 2004). Both milk types showed non significant (P ≥
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0.05) difference with respect to acidity and pH. The reduction in the total solids of lupin milk
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might be due to the removal of the dry matter during milk processing as it has been previously
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reported that debittering reduced the total solids of lupin milk (Erbas, 2010; Jimenez-Martinez et
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al., 2001). The chemical composition of milk, especially the contents of protein, fat, calcium and
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pH, has a major influence on several aspects of cheese manufacture, especially rennet
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coagulability, gel strength, curd synergistic, and hence cheese composition and cheese yield
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(Fox, & Cogan, 2004).
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3.2 Lupin-based cheese formation and yield
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Substituting cow milk at 25, 50, and 75 mL/ 100 mL for lupin milk was observed to form
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cheese, without major effect on the curd formation compared to control cheese of pure cow milk
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(Fig. 2). Moreover, a remarkable increase in cheese yield was observed with increasing
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supplementation level of lupin milk. The mean value of cheese yield of pure cow milk was
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found to be 0.925 kg/ 5 kg milk and then gradually increased (P ≤0.05) to 0.975 kg/ 5 kg, 1.0
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kg/5 kg, and 1.12 kg/5kg with increasing lupin milk level to 25, 50 and 75 mL/100 mL,
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respectively. The enhancement of cheese yield could be attributed to the higher content of fat
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and protein in lupin milk compared to cow milk as well as low losses of total soluble solids in
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whey. In contrast, cheese yield decrease was reported in whey protein concentrate fortified milk
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cheese (Lobato-Calleros, Reyes-Hernandez, Beristain, Hornelas-Uribe, Sanchez-Garcia, &
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Vernon-Carter, 2007) and sesame protein isolate supplemented fresh cheese (Lu, Schmitt, &
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Chen, 2010). On the other hand, whey produced during cheese making was ranged from 2.5 to 3
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liter, with a lower level in cheese made from pure cow milk. This may likely be due to higher
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casein level in pure milk which leads to a slow rate of exclusion of whey or may be to Ca++
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activity which is an important factor in determining the rate of whey drainage. Additionally,
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slightly lower pH of lupin milk and the mixtures thereof could also contribute to the higher whey
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drainage of lupin based cheeses compared to the control cheese. Both higher casein
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concentration and pH were reported to decrease the curd syneresis and whey drainage (Daviau et
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al., 2000).
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3.3 Biochemical characteristics of lupin-based cheese
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Table 3 shows the biochemical characteristics of fresh soft cheese made with cow milk
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with and without lupin milk. The concentrations of protein, fat, total solids and acidity were
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significantly (P ≤0.05) increased with increasing lupin milk level in fresh soft cheese compared
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to pure milk fresh cheese. On the other hand, ash and pH showed significant (P ≤0.05) reduction
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as the level of lupin milk increased. Higher protein content (11.96 g/100 g) was observed when
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cow milk was mixed with 75 mL/100 mL lupin milk compared to that made from pure cow milk.
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The protein content of the soft cheese under investigation is slightly lower than that reported for
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Chinese traditional soy cheese known as sufu or furu (Ahmad, Yang, Ning, & Randhawa, 2008).
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As the level of lupin milk of fresh cheese increased over 25 mL/100 mL, fat content was
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significantly (P ≤0.05) increased which improved the quality of lupin containing cheese. Fats
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are known to play a major role in the rheology and texture as well as flavour of the product by
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acting as a source of fatty acids which in turn may be catabolized to flavour compounds, e.g.,
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methyl ketones, esters, thioesters and lactones (Collins, McSweeney, & Wilkinson, 2003). Total
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solids content of fresh cheese was significantly (P ≤0.05) increased with increase in the level of
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lupin milk. This might be due to lower water content of lupin-based cheese compared to that of
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control cheese as the previous result showed that the whey exclusion of lupin-based cheese was
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higher compared to that of the control cheese. The increase in total solids in lupin-based cheese
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could also be attributed to higher protein and fat contents in lupin milk compared to cow milk.
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The results obtained were within the range reported for white cheese prepared from cow milk (El
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Owni & Hamed, 2009). As the level of lupin milk increased in fresh cheese a significant (P
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≤0.05) reduction in ash content was observed compared to that of pure milk which could be
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attributed to the lower amount of ash in lupin milk. For unknown reasons, the acidity of fresh
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cheese was significantly (P ≤0.05) increased when the level of lupin milk was increased.
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Significantly (P ≤0.05) low acidity value was observed in soft fresh cheese made from pure cow
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milk, compared to that obtained for cheese containing lupin milk. This could be attributed to
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change in the proportion of lactose which is converted by lactic acid bacteria to lactic acid. The
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complete and rapid metabolism of lactose in cheese curd is essential for the production of good
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quality cheese since the presence of a fermentable carbohydrate may lead to the development of
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an undesirable secondary flora (Fox, Guinee, Cogan, & McSweeney, 2000). Since low acidity
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curd normally contains more moisture than high acidity curd, one might expect more syneresis
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and higher whey drainage of low acidity curd. The pH of fresh soft cheese was significant (P
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≤0.05) decreased when the level lupin milk was increased in the cheese compared to that of fresh
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cheese prepared from pure milk. The reduction of the pH could be attributed to the increased
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acidity of the cheese with higher level of lupin milk. Since the increase in the acidity is always
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contradictory with the pH.
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Table 4 shows the impact of maturation for 75 days on biochemical characteristics of pure
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milk cheese and that manufactured by addition of lupin milk of different levels. Mature cheese
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prepared from a mixture of pure milk and lupin milk contained higher (P ≤0.05) amount of
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protein compared to that prepared from pure milk. When compared to fresh soft cheese, a 14
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remarkable reduction in the protein content was observed in all types of mature cheese. Despite
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inactivation of the enzymes by heating, the reduction in protein content could be due to
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hydrolysis of the protein in cheese by the residual chymosin, indigenous proteinases and/or
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microbial proteinases from starter and non-starter microorganisms. Primary proteolysis of cheese
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proteins is mainly the result of the action of endogenous proteinases (i.e., plasmin and somatic
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cell proteinases) and residual coagulant. However, proteinases from starter and nonstarter
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microorganisms can also be active in the degradation of cheese proteins and peptides. Lactic acid
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bacteria are weakly proteolytic, but possess a very comprehensive range of proteinases and/or
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peptidases capable of hydrolyzing casein-derived peptides to small peptides and amino acids.
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Proteolysis is the most complex, and in most cases, the most important biochemical event which
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occurs during cheese ripening and it contributes to the development of cheese texture as well as
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flavour (Sousa, Ardo, & McSweeney, 2001). Fat content of mature control cheese as well as that
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contain 25 mL/100 mL lupin milk was slightly increased compared to fresh soft cheese.
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However, the fat content of mature cheese significantly (P ≤0.05) decreased with increase in
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lupin milk level compared to that of cheese prepared from pure milk. Ash content of mature
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cheese significantly (P ≤0.05) increased at all levels of lupin milk compared to fresh cheese. The
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increases in the ash content during storage might be due to the decrease in moisture content as
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well as absorption of salt by cheese curd. Total solids increased in mature cow milk cheese
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compared to the fresh one. However, mature cheese prepared from a mixture of lupin and cow
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milk had low total solids compared to fresh cheese. Moreover, as the level of lupin milk
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increased, total solids of mature cheese were decreased. The acidity of mature cheese prepared
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from pure milk was significantly (P ≤0.05) increased compared to fresh cheese. The
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enhancement of the acidity of pure milk cheese might be due to the formation of lactic acid by
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lactic acid bacteria during fermentation of lactose. However, addition of lupin milk significantly
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(P ≤0.05) reduced the acidity of mature cheese compared to fresh one. This might be due to the
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lack and/or low concentration of fermentable sugar (lactose) in lupin containing cheeses. The
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complete and rapid metabolism of the lactose and its constituent monosaccharides in cheese curd
324
is essential for the production of good quality cheese since the presence of a fermentable
325
carbohydrate may lead to the development of an undesirable secondary flora during ripening.
326
The acidity depends primarily on type and quantity of starter added, pre-acidification applied,
327
temperature and duration of acid development. The pH of lupin containing mature cheese was
328
significantly (P ≤0.05) higher than that of fresh one. As the level of lupin milk increased the pH
329
of mature cheese gradually (P ≤0.05) decreased and reached 6.10 at 75 mL/100 mL level but still
330
higher than that of fresh cheese. The rises of the pH of cheeses during ripening could be
331
attributed to the formation of alkaline nitrogenous compounds (such as ammonia) and/or
332
catabolism of lactic acid and thus reduce the acidity of the cheeses.
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3.4 Microbial analysis lupin-based cheese
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Table 5 shows the microbial evaluation of cheeses made from cow milk mixed with
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different levels of lupin milk. The total bacterial count is within the range that naturally exists in
337
milk containing foods. Compared to pure milk cheese, cheese made with 75 mL/100 mL lupin
338
milk had lower bacterial count followed by that made of 25 mL/100 mL lupin milk. Higher
339
bacterial count was observed when cow milk substituted with 50 mL/100 mL for lupin milk.
340
Other microorganisms such as fungi, mould, E. coli, and Salmonella were not detected. Similar
341
results of bacterial count as well as absence of fungi, mould, E. coli and salmonella were also
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observed in lupin milk before incorporation into cheeses (data not shown). The absence of such
343
microorganisms in both lupin milk and the cheese made thereof may be due to the extraction
344
method used for the preparation of lupin milk in which clean water was used for the extraction as
345
well as pasteurization of the milk used in the cheese making. Similar observation was observed
346
in vegetable-based dairy products (Aminigo, Metzger, & Lehtola, 2009). In addition to the use of
347
high temperature, high level of NaCl (1.4–2.6 mol/L) when added to the cheese milk will control
348
the growth of the indigenous microflora (Mohamed Ahmed et al., 2010).
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3.5 Sensory characteristics of lupin-based cheese
The sensory characteristics of fresh soft cheese made using different levels of lupin milk
352
are shown in Table 6. The cheese prepared by adding different levels of lupin milk was not
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significantly (P ≥ 0.05) differed from that of pure milk with respect to taste and texture except
354
that prepared from 75 mL/100 mL lupin milk which had lower scores of taste and texture. Higher
355
taste score was obtained for cheese made with 25 mL/100 mL lupin milk and the lower one was
356
observed for cheese made with 75mL/100 mL lupin milk. Moreover, no significant (P ≥ 0.05)
357
differences were observed in the color of all types of cheese. Addition of lupin milk at the level
358
of 50 and 75 mL/100 mL resulted in low score of the color of the cheese compared to addition of
359
25 mL/100 mL lupin milk and that of pure milk. Incorporation of lupin milk in cheese
360
significantly (P ≤0.05) improved the flavour of it. Higher score of flavor was obtained for
361
cheese made with 75 mL/100 mL lupin milk whereas the lower one was observed in cheese
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prepared from pure milk. This probably could be due to the degradation of carbohydrates to
363
lactic acid and release of flavour components such as acetaldehyde or decomposition of fat into
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volatile fatty acid. There are significant (P ≤0.05) differences in the overall acceptability
365
between pure milk cheese and those prepared by adding 50 and 75 mL/100 mL lupin milk. Panel
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test showed that fresh cheese prepared from 50 and 75 mL/100 mL lupin milk are less acceptable
367
compared to that of pure milk and that prepared by mixing 25 mL/100 mL lupin milk. Strikingly,
368
incorporation of lupin milk in the cheese at low concentration (25 mL/100 mL) does not caused
369
any defects, and even slightly improved, the sensory characteristics of fresh soft cheese
370
compared to the control cheese made of pure cow milk. Whereas, high concentration (75 mL/
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100 mL) of lupin milk significantly (P ≤0.05) affect the sensory characteristics of fresh soft
372
cheese compared to the control cheese and that made of 25 mL/100 mL lupin milk.
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Table 7 shows the sensory characteristics of mature cheese prepared from cow milk with or
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without lupin milk. It is noted that some sensory characteristics such as taste, texture and overall
375
acceptability in cheese made with 25 mL/100 mL lupin milk were improved during maturation
376
except colour and flavour compared to fresh cheese. For mature cheese of pure milk the taste,
377
color and overall acceptability were improved with exception of flavor and texture. Generally,
378
the overall acceptability of all types of cheese was enhanced during maturation. The results
379
obtained agree with the findings of Tarakci and Kucukoner (2006) who reported that the overall
380
acceptability of cheese was increased during ripening. The results also showed that there are
381
significant (P ≤0.05) differences in the scores of taste, texture, flavor and overall acceptability
382
between all types of cheese including the control one. In lupin containing cheese, the increase of
383
lupin milk concentration significantly (P ≤0.05) reduced the scores of taste, texture and overall
384
acceptability of the cheese but it significantly (P ≤0.05) improved the flavor. Lowest scores of
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taste, texture, color, and overall acceptability among all cheeses including the control one were
386
observed in cheese made with 75 mL/100 mL of lupin milk. Interestingly, incorporation of lupin
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milk in the cheese at low concentration (25 mL/100 mL) significantly (P ≤0.05) enhanced the
388
taste, texture, flavor, and overall acceptability of the final mature cheese compared to the control
389
cheese. With an exception of the flavor, at higher concentrations (50 and 75 mL/100 mL) of
390
lupin milk in the cheese these all sensory attributes were significantly (P ≤0.05) decreased
391
compare to those of control cheese. Overall, substitution of cow milk at 25 mL/100 mL for lupin
392
milk significantly enhanced most of the sensory attributes of final mature cheese, with slight
393
defect in the color compared to the control cheese of pure cow milk. The sensory characteristics
394
of a cheese at the time of its consumption reflect the quality of milk from which it was produced,
395
the processes used for the production and the physical and chemical changes that may occurred
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during maturation (Delahunty & Drake, 2004).
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4. Conclusion
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The results of this study demonstrated that lupin milk containing cheese at low substitution
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of cow milk (25 mL/100 mL) for lupin milk has close resemblance to that of cow milk cheese in
401
some nutritional and sensory characteristics. Substitution of cow milk at 25 mL/100 mL for lupin
402
milk can be used for the production of white soft cheese without causing any major changes in
403
quality attributes of both fresh and mature cheese. Moreover, incorporation of lupin milk in
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cheese manufactured resulted in cost saving and improvement of the nutritional value.
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References
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Johnson, S.K., Chua, V., Hall R.S., & Baxter A.L. (2006). Lupin kernel fibre foods improve
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bowel function and beneficially modify some putative faecal risk factors for colon cancer in
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Lee, Y.P., Mori, T., Sipsas, S., Barden, A., Puddey, I., Burke, V., Hall R., & Hodgson, J. (2006).
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J.E., & Vernon-Carter, E.J. (2007). Microstructure and texture of white fresh cheese made
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Lu, X., Schmitt, D., & Chen, S. (2010). Effect of sesame protein isolate in partial replacement of
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milk protein on the rheological, textural and microstructural characteristics of fresh cheese
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Martins, J.M., Riottot, M., de Abreu, M.C., Viegas-Crespo, A.M., Lança, M.J., Almeida, J.A.,
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Freire, J.B., & Bento, O.P. (2005). Cholesterol-lowering effects of dietary blue lupin
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activity of a milk-clotting enzyme from Solanum dubium seeds and its enzymatic action on
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Pastor-Cavada, E., Juan, R., Pastor, J. E., Alaiz, M., & Vioque, J. (2009). Analytical nutritional
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characteristics of seed proteins in six wild Lupinus species from Southern Spain. Food
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during cheese ripening. International Dairy Journal, 11, 327–345. Vecerek, V., Suchy, P., Strakova, E. & Machacek, M. (2008). Nutritive composition of seeds of
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the lupin varieties registered in the Czech republic. In: J.A. Palta and J.B. Berger (eds).
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Tarakci, Z., & Kucukoner, E. (2006). Changes in physicochemical, lipolysis and proteolysis of
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vacuum packed Turkish Kashar cheese during ripening. Journal of Central European
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Agriculture, 7, 459-464.
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Table 1. Chemical composition of the local variety of lupin seeds (dry weight) and
509
other lupines.
(g/100 g)
Local variety
Other Lupins*
(Rubatab) 46.3(±2.31)
Fat
10.4(±1.14)
Carbohydrate
27.1(±1.26)
Ash
1.6(±0.10)
Moisture
4.6(±0.11)
Fiber Mineral composition (g/100 g) Ca Na
P
4.5-8.0 ND
3.77-5.2 ND
10.0(±1.15)
12.8-16.9
0.4(±0.03)
0.27-0.48
0.01(±0.02)
ND
0.85(±0.1)
ND
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33.2-43.7
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Chemical composition
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0.86(±0.09)
*Vecerek et al. (2008). ND; not detected
511
Values are means of triplicate samples (±SD)
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0.55-0.92
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Parameter
Lupin milk
Cow milk
Protein (g/100 g)
4.90 (±0.10)
Fat (g/100 g)
Concentration of lupin milk (mL/100 mL) 50
75
3.90 (±0.08)
4.10 (±0.06)
4.50 (±0.09)
4.70 (±0.10)
5.00 (±0.12)
3.80 (±0.13)
4.20 (±0.08)
4.40 (±0.08)
4.80 (±0.12)
Acidity (% lactic acid)
0.30 (±0.01)
0.20 (±0.01)
0.23 (±0.04)
0.25 (±0.05)
0.26 (±0.05)
Total solids (g/100 g)
11.20 (±0.07)
12.60(±0.11)
12.30 (±0.18)
12.0 (±0.13)
11.40 (±0.10)
pH
6.30 (±0.10)
6.40 (±0.06)
6.38 (±0.05)
6.35 (±0.06)
6.31(±0.02)
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Values are means of triplicate samples (±SD)
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Table 2. Biochemical composition of lupin milk, cow milk and mixtures of cow and lupin milk
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Table 3. Biochemical composition (g/100 g) of fresh soft cheese made from cow milk supplemented with lupin milk of different levels. Concentration of lupin milk (mL/100 mL) 25
50
Protein
6.66c (±0.07)
8.90b(±0.12)
8.96b(±0.10)
11.96a(±0.11)
Fat
10.00c(±0.10)
10.00c(±0.11)
12.00b(±0.15)
12.33a(±0.18)
Ash
2.49a(±0.05)
1.13b(±0.06)
0.89c(±0.01)
0.69d(±0.03)
Total solids
38.23c(±0.64)
40.40b(±0.69)
40.63b(±0.61)
42.30a(±0.76)
Acidity (% lactic acid)
0.18d(±0.01)
0.80c(±0.03)
1.66b(±0.04)
2.66a(±0.08)
pH
6.43a(±0.09)
5.16d(±0.08)
5.46b(±0.10)
5.23c(±0.06)
Values are means of triplicates (±SD). are significantly different at (P ≤0.05).
a-d
SC
:Means average in the same raw with different letters
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Parameter
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519 520
Table 4. Biochemical composition (g/100 g) of mature soft cheese made from cow milk supplemented with lupin milk of different levels. Concentration of lupin milk (mL/100 mL) 25
50
Protein
3.03d(±0.04)
4.23b(±0.05)
4.70a(±0.07)
4.11c(±0.04)
Fat
11.66a(±0.13)
11.33b(±0.09)
10.33d(±0.11)
10.66c(±0.07)
Ash
3.19a(±0.06)
1.09d(±0.01)
1.46c(±0.03)
1.56b(±0.04)
Total solids
39.80a(±0.55)
39.20b(±0.23)
38.33c(±0.32)
37.80d(±0.19)
Acidity (% lactic acid)
0.80a(±0.03)
0.70b(±0.02)
0.40d(±0.01)
0.50c(±0.01)
pH
6.70a(±0.05)
6.63b(±0.04)
6.30c(±0.04)
6.10d(±0.03)
Values are means of triplicates (±SD). are significantly different at (P ≤0.05).
a-d
SC
:Means average in the same raw with different letters
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523
75
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0
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Parameter
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Table 5. Microbial analysis of soft cheese made from cow milk supplemented with different levels of lupin milk Test
Bacterial count 526
Concentration of lupin milk (mL/100 mL) 0
25
50
3.3× 106 cfu/ml
2.6× 106 cfu/ml
5.6× 106 cfu/ml
Other microbes such as Fungi, Yeast, E. coli and Salmonella were not detected.
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527
30
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1.3× 106 cfu/ml
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528 529
Table 6. Sensory characteristics of fresh soft cheese made from cow milk supplemented with different levels of lupin milk Concentration of lupin milk (mL/100 mL) 25
50
75
Taste
3.50a(±0.04)
3.58a(±0.08)
3.50a(±0.05)
2.91b(±0.03)
Texture
3.50a(±0.03)
3.50a(±0.04)
3.58a(±0.09)
2.91b(±0.01)
Color
3.33a(±0.07)
3.33a(±0.09)
3.16a(±0.08)
3.16a(±0.10)
Flavor
2.75c(±0.02)
3.16b(±0.04)
3.16b(±0.06)
3.58a(±0.09)
Overall acceptability
3.00a(±0.03)
3.08a(±0.05)
2.66b(±0.01)
2.25c(±0.04)
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Values are means of 12 samples (±SD). a-c :Means average in the same raw with different letters are significantly different at (P ≤0.05). Hedonic scale (1–5, 1 = poor, 5 = excellent).
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Quality attribute
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Table 7. Sensory characteristics of mature soft cheese made from cow milk supplemented with different levels of lupin milk
0
25
50
Taste
3.75b(±0.02)
4.16a(±0.04)
3.08c(±0.02)
2.41d(±0.03)
Texture
3.50b(±0.03)
3.58a(±0.04)
3.25c(±0.01)
2.25d(±0.04)
Color
4.00a(±0.10)
3.25b(±0.07)
3.33b(±0.06)
3.16b(±0.08)
Flavor
2.75d(±0.01)
3.08c(±0.05)
3.58b(±0.05)
3.83a(±0.02)
Overall acceptability
3.75b(±0.03)
4.16a(±0.09)
3.08c(±0.04)
2.41d(±0.01)
544
EP
543
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542
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539
541
75
Values are means of 12 samples (±SD). a-d :Means average in the same raw with different letters are significantly different at (P ≤0.05). Hedonic scale (1–5, 1 = poor, 5 = excellent).
538
540
SC
Quality attribute
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Concentration of lupin milk (mL/100 mL)
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545 546
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Fig.1. Preparation of lupin milk from the seeds of Lupinus albus
547 548 549 33
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Fig. 2. White soft cheese produced using cow milk supplemented with lupin milk at different concentrations of 0 mL/100 mL (A), 25 mL/100 mL (B), 50 mL/100 mL (C), and 75 mL/100 mL (D).
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Physicochemical, microbiological and sensorial properties soft cheese made of mixture of cow and lupin milk were evaluated.
Lupin milk based cheese at low substitution (25 mL/100 mL) for lupin milk
sensory characteristics.
Substitution of cow milk at 25 mL/100 mL for lupin milk could be used without major changes in the quality of final cheese.
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has close resemblance to that of cow milk cheese in terms of biochemical and
Incorporating lupin milk into cheese could save money as well as improve the
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nutritional value of such product.
ACCEPTED MANUSCRIPT Research Highlight
Quality attributes of cheese made of mixture of cow and lupin milk were evaluated.
Substitution of cow milk at 25 mL/100 mL for lupin milk produced good quality cheese.
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Incorporating lupin milk into cheese could save money.
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