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Spray drying strategy for encapsulation of bioactive peptide powders for food applications
Advanced Powder Technology xxx (xxxx) xxx
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Perspective article
Spray drying strategy for encapsulation of bioactive peptide powders for food applications q Yong Wang a, Cordelia Selomulya a,b,⇑ a b
Department of Chemical Engineering, Monash University, 18 Alliance Lane, Clayton, VIC 3800, Australia School of Chemical Engineering, UNSW, Sydney, NSW 2052, Australia
a r t i c l e
i n f o
Article history: Received 1 July 2019 Received in revised form 28 October 2019 Accepted 30 October 2019 Available online xxxx Keywords: Bioactive peptide Spray drying Encapsulation Food proteins
a b s t r a c t Spray drying is commonly used to manufacture food and dairy powders at industrial scale. Bioactive peptides, as hydrolysis products from various food proteins, are gaining interest due to their biological functionality including immunomodulation, antimicrobial, and antioxidant properties. This article reviews several bioactive peptides/hydrolysates from different food sources, focusing on their processing and final powder characteristics. Additionally, we propose a strategy for encapsulation of food peptide/hydrolysate via spray drying. The identification of suitable drying parameters and/or formulation could help overcome the limitations associated with the current processing of food peptides. Ó 2019 The Society of Powder Technology Japan. Published by Elsevier B.V. and The Society of Powder Technology Japan. All rights reserved.
1. Introduction The application of peptides and protein hydrolysate to food is at the forefront of innovation for modern nutrition and represents a promising approach for efficient delivery of bioactive compounds. In line with this, various peptides for food applications have recently attracted interest. Bioactive peptides are specific protein fragments that are inactive within the sequence of the parent protein, but may exert various physiological functions after release during digestion [1]. In general, peptide refers to a more uniform protein fragment, while hydrolysate is a complex proteolytic mixture. However, unlike in the pharmaceutical industry, food peptide or protein hydrolysate do not have a strict distinction. Therefore, both terms will be used here interchangeably. Biological properties reported for peptides derived from food proteins include anti-aging, anti-cancer, anti-cariogenic, antidiabetic, anti-hypertensive, anti-microbial, anti-oxidative stress, anti-inflammatory, cholesterol lowering, growth enhancing, immunomodulatory, mineral binding, radical scavenging, regulation of glucose and insulin homeostasis, and satiety regulating [1–5]. In addition, the application of peptides into food is beneficial for populations with special nutritional needs such as infants, q This is the perspective article submitted for Virtual Special Issue: the 30th Anniversary Collection of Advanced Powder Technology. ⇑ Corresponding author at: Department of Chemical Engineering, Monash University, 18 Alliance Lane, Clayton, VIC 3800, Australia. E-mail addresses: [email protected], cordelia.selomulya@unsw. edu.au (C. Selomulya).
athletes and the elderly, due to its ease of digestion [6]. Thus the development of functional foods or nutraceuticals fortified with peptide is increasing [7]. Generally, food peptides require spray drying to be converted into powder for further applications. High cost of production, poor solubility of purified peptides powders, and consumer acceptance are among some of the challenges for commercialization of foodderived bioactive peptides [1,8]. Thus, a systematic design of spray drying trials is needed to optimize the process parameters to preserve the desirable attributes of the peptide. The effect of drying on the functional properties such as bioactivity and bioavailability of peptides needs to be considered. It is also necessary to account for the state of peptide particles after drying, including moisture content, shelf life, solubility, ease of use, taste/odour control, and others [9]. Since spray drying is one of the most energy consuming and expensive steps in the production of protein hydrolysates, care must also be taken to balance the energy consumption of the drying process, inlet/outlet temperature selection, and the final quality of the product. [10]. In industrial production, often a compromise must be made between the yield of hydrolysate that can be achieved versus the amount of water that must be removed to obtain a dry final product, e.g. adding less water during hydrolysis to minimize drying cost [10,11]. There are a number of studies on peptide that mainly focus on the amino acid composition [12–14], while microencapsulation of food peptide is also of interest [15]. Although with the same amino acid composition, the peptide functionality may vary greatly depending on the powder properties. Peptide is more fragmented
https://doi.org/10.1016/j.apt.2019.10.034 0921-8831/Ó 2019 The Society of Powder Technology Japan. Published by Elsevier B.V. and The Society of Powder Technology Japan. All rights reserved.
Please cite this article as: Y. Wang and C. Selomulya, Spray drying strategy for encapsulation of bioactive peptide powders for food applications, Advanced Powder Technology, https://doi.org/10.1016/j.apt.2019.10.034
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than protein, which makes the drying process more complicated if carrier is added into formulation and might interact with the bioactive peptides [9]. Another key consideration in encapsulation of peptides via spray drying is how well these thermally labile materials resist heat denaturation by hot air [16]. Although it is a common procedure to process protein-based food into powders, to the best of our knowledge, only limited studies have been done on developing a spray drying strategy to maximize bioactivity and functionality of food peptides, especially in their pure or concentrated forms. Here, we briefly summarized recent studies on the spray drying of food peptides, and propose areas for further investigation to achieve better quality and expand the application of food peptide powders. 2. Current research on food peptide drying The existing literature on food protein peptide or hydrolysate drying can be divided into three categories: (i) Direct drying of pure peptide [17,18]; (ii) Co-drying with processing aids (e.g. maltodextrin or starch), but not necessarily encapsulated even with claims of microencapsulation in some cases [19–21]; (iii) Microencapsulation (mainly using wall materials to encapsulate peptide, but may also use peptide as wall material to encapsulate other components) [9,22,23]. The main purpose of the first two categories is to manufacture powders rich in peptide for use as food or nutraceutical ingredients, while the main purpose of last category is on maintaining bioactivity or masking the smell/taste of the peptide. 2.1. Dairy peptide As major components of dairy protein, whey and casein peptides are spray dried and sold as powdered ingredients. Ma et al. compared the effects of spray drying and freeze drying on bioactivity, taste, and hygroscopicity of whey hydrolysate, and found that spray-dried pure whey hydrolysate showed similar bitter taste, hygroscopicity, and solubility compared to freeze-dried whey hydrolysate [17]. Contreras et al. compared the spray-dried and freeze-dried casein peptides, and found that the angiotensin converting enzyme (ACE)-inhibitory activity and the antihypertensive properties of casein hydrolysed products were maintained after spray drying or freeze drying [24]. Moreover, the spray-dried casein hydrolysate powder still showed their antihypertensive effects when added into yoghurts, indicating their potential as food ingredient [24]. The allergic properties of bovine milk proteins could be reduced by enzymatic hydrolysis [25]. But their relatively high cost and bitter taste may restrict the use of milk peptide in infant milk formula. However, recent studies demonstrated the stability of spray-dried whey hydrolysate powders throughout thermal and spray drying processes during infant milk formula manufacture, with the potential to lower drying cost by decreasing liquid viscosity, and increasing storage stability by reducing lactose crystallization [26,27]. On the other hand, the high hygroscopicity of whey hydrolysate powders may decrease the storage stability of infant milk formula by lowering the glass transition temperature during storage [28]. In addition, spray-dried whey protein hydrolysate were more susceptible to sticking due to the low glass transition temperature, also with lower surface polarity when compared to intact proteins, which need to be taken into consideration when using whey hydrolysate to fortify other dairy products [29,30]. Drusch et al. found that both casein hydrolysate and whey hydrolysate showed higher surface activity than casein or whey protein at the air-water interface, because of lower molecular weight and more ionisable groups. Thus the hydrolysed milk
proteins can be used to alter the surface composition of the spray-dried particles [31]. Better encapsulation of fish oil from oxidation was shown with whey hydrolysate as wall material, compared to b-lactoglobulin, reportedly due to higher accessibility of antioxidant amino acids from wall material due to enzymatic hydrolysis [22]. Microencapsulation of whey hydrolysate with whey protein and sodium alginate did not change the immunomodulatory activity, but could lower their hygroscopicity and solubility, and provide bitterness-masking effect [17]. Maltodextrin-encapsulated whey protein hydrolysate was significantly less hygroscopic than without encapsulation [32], while microencapsulation also helped to improve gastrointestinal stability of the whey protein hydrolysate [23]. For casein peptide, the bitter flavour can be reduced by spray drying with different carriers including gelatine and soy protein isolate, gum Arabic, and maltodextrin [33–36]. However, the rate of dissolution of casein hydrolysate in water may decrease significantly due to the introducing of wall material in encapsulation process [34]. 2.2. Chicken peptide Chicken meat could be hydrolysed and spray-dried into hydrolysate powders. Higher inlet temperature in spray drying resulted in lower powder moisture and bulk density, as well as spherical shape of hydrolysate powder [20]. In addition, high inlet air temperature (170–200 °C) in spray drying may also bring advantages like higher antioxidant activity of chicken meat hydrolysate powder as evidenced in precious study [37]. On the contrary, lower drying temperature (120–160 °C) combined with low feed flow rate could minimize the difference between inlet and outlet temperature in spray drying, in order to achieve high product recovery for chicken peptides [37]. Maltodextrin and gum Arabic could be used as carrier to improve the drying performance of chicken meat peptide by reducing the stickiness and hygroscopicity of hydrolysate powders, and also increasing their glass transition temperature [20,38]. Although freeze dried chicken skin hydrolysate showed higher water solubility, emulsion stability, and water holding capacity than the spray-dried powders, spray drying is still preferable due to its high production efficiency [39]. 2.3. Fish peptide Fish meat, especially the by-product of fillet, could be spraydried into powder for application in food or as animal feed. Spray-dried Black Tilapia fish hydrolysate powders showed high in vitro digestibility (up to 92.1%) at both inlet temperature (150 °C or 180 °C), and lower drying inlet temperature (150 °C) was preferable to achieve higher protein efficiency ratio in animal feed [21]. In another study, an inlet temperature of 185 °C was used to dry the protein hydrolysate of different variety of fish including Mackerel, White Fish, Blue Whiting, and Sprat [18]. The bitter flavours of dried fish hydrolysate powders were found to be relatively mild compared with the hydrolysates from milk proteins, showing their potential to be added into food matrix. Spraydried Sauvage fish protein hydrolysate powder (inlet temperature 180 °C) showed better oil absorption capacity and emulsion activity when the hydrolysis was finished by alkali or acid instead of enzyme, but the enzyme-hydrolysed powder has higher water solubility than the alkali or acid hydrolysis powders [40]. Pretreatment with ethanol could slow down the non-enzymatic browning of spray-dried fish hydrolysate powders (inlet temperature 180 °C), and also mask the bitter taste [41]. It has been suggested that despite the extensive studies of the hydrolysis process, further studies on the drying and storing of fish hydrolysates are necessary for industrial application [11,18].
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2.4. Other food peptides There are several features that may limit the application of plant proteins, including low solubility. Research on plant protein hydrolysate has the potential to bring new applications into the food industry. Spray-dried soy protein hydrolysate powder showed significantly higher solubility than soy protein isolate around pH 4.5, which is the isoelectric point of soybean protein. Soy hydrolysate also showed higher foaming capacity and lower viscosity, while retaining gel-forming ability, which is favourable for application in meat product [42]. Wang et al. investigated the effects of inlet drying air temperature (140–200 °C), feed flow rate (3– 12 mL/min), and solids concentration (25–40%) on the properties of spray-dried soy protein peptide powders, and found an optimized spray drying condition for soy peptide (160 °C inlet temperature, 6 mL/min feed rate, and 35% solid concentration) based on powder characteristics including colour, morphology, density, and antioxidant capacity [43]. Using maltodextrin as carrier, spray-dried flaxseed protein hydrolysate powder could not only show good solubility (92–96%), but also retain high antioxidant activity [44]. Egg white protein hydrolysate powder exhibited similar solubility and antioxidant activities when dried by either spray drying or freeze drying. However, spray drying could lower the emulsifying and foaming properties of egg white protein hydrolysate powder when compared with the freeze-dried powder, which might be caused by the low hydrophobicity of spray-dried peptide [45]. In another study, spray-dried egg white protein peptides (inlet temperature 180 °C) showed high in vitro angiotensin converting enzyme (ACE) inhibitory activity, which might be beneficial in preventing hypertension, heart failure, myocardial infarction, and diabetic nephropathy [46]. In particular, spray-dried egg white peptide Arg-Val-Pro-Ser-Leu (RVPSL) showed the potential as a functional food ingredient with highest ACE-inhibitory activity [47], while peptide Asp-His-Thr-Lys-Glu (DHTKE) had the best oxygen radical absorbance capacity [48]. Spray-dried mussel meat protein hydrolysate powders had a higher storage stability in comparison to chicken breast hydrolysate powder [49]. The addition of maltodextrin or gum Arabic as carrier might increase the glass transition temperature of mussel meat protein hydrolysate powders (from 64 °C to 69–97 °C) and decrease water adsorption, which is beneficial for storage stability [49]. The spray-dried mussel meat protein hydrolysate with maltodextrin carrier showed potential to be used as umami seasoning in instant noodles [50].
3. Drying strategy for food peptide Based on the analysis of existing data and our previous drying research experience, we recommend the following parameters to be considered when spray drying food protein peptide/hydrolysate into powders (shown in Fig. 1). Before drying, the hydrolysis process and the characteristics of the peptide/hydrolysate need to be considered. Several factors in the hydrolysis process affect the properties of hydrolysate, which in turn affects the drying process. The degree of hydrolysis, molecular weight distribution of the hydrolysate, and the amino acid composition may affect or even to some extent determined the water-binding ability of the peptide molecules [51]. This further affects the rheological property, surface charge, heat tolerance, and peptide interaction of the suspension before drying [52]. Other factors are easier to control, such as the moisture content and pH of the suspension, but also have a large impact on the subsequent drying process. Due to the variation in the functionality of peptides, the separation and purification steps can affect their
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bioactivity [4,53], which is crucial when claiming functionality of the ingredients in food. During drying, various parameters need to be adjusted carefully to accommodate the drying requirements of the peptide. Optimum drying conditions and tailored matrix formulations are required to avoid severe heat damage leading to loss in bioactivity [54]. Similar to the spray drying of other powders, the critical parameters that can be controlled are inlet temperature, outlet temperature and feeding rate [16]. Some operating parameters are determined by the type of spray dryer, such as evaporation efficiency and air flow, such that suitable conditions need to be determined for a specific dryer (e.g. drying aids or carriers) where computational modelling may help [55]. Unlike the drying of pure peptide, more factors need to be taken into consideration for encapsulation, including the carrier type, ratio between peptide and carrier, and pre-treatments, etc [16,56,57]. In addition, the understanding of droplet drying kinetics via single droplet drying may provide fundamental understanding of changes that occur in the peptide suspension during drying. Our works have demonstrated the use of single droplet drying to provide insight into micro-structural changes during drying [58,59], in addition to drying kinetics data that can be used in modelling the drying process including water vapor, drying rate, shrinking, etc [54,60,61]. Novel spray drying methodology, such as nano spray drying, developed to enhance powder solubility of pharmaceutical peptides, might also help to preserve the functionality of food peptide by reducing the droplet size and the drying duration [62]. After drying, the analysis of the powder is also critical. The physical characteristics of peptide powder, including particle size distribution, moisture content, morphology, and others, are relevant to properties such as solubility and shelf stability. The formation of porous granules can improve solubility in water, while the physical structure of the powder are correlated with the digestibility and peptide release [35,63].The thermal properties of peptide powder, such as glass transition temperature, together with other physical properties, will help to quantify flowability, stickiness, and stability of the peptide powder [64]. Both the functionality and sensory properties of peptide powders determine their applications in food matrix. In addition to solubility, emulsifying and foaming capacities will also determine the application of peptide powder [65]. Water activity and storage stability not only affects the peptide powder itself, but also affects the whole formulated food [26,66,67]. In some cases, the peptide powder may induce bitter taste and undesirable odour [9]. This requires consideration of the appropriate drying condition to mask the negative effect to food matrix. On the contrary, there are some cases where the peptide powder may introduce pleasant taste like umami, especially from those generated from fish and seaweed [1,68]. Bioavailability needs to be assessed after drying to ensure the bioactive peptides can still function normally [9,69]. The last thing to note is the complex composition of food matrix, where a comprehensive analysis is needed since any change can bring unexpected effect on flavour or texture.
4. Opportunities for research on food peptide powders In the previous section, although we try to list most of the technical areas to be considered in the peptide drying, there are some areas worthy of further investigation. These include how to maintain the bioactivity of peptide during the drying process, how to optimize the drying parameters to generate peptide powders with desired properties, and how to render peptide powders more compatible with the food matrix.
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Fig. 1. An overview of relevant process parameters and properties in the spray drying of food protein peptide/hydrolysate.
4.1. Drying protection The fundamental issue is how to develop methods for drying food peptide/hydrolysate without introducing unexpected changes in relevant powder properties. At the same time, consideration should be made regarding the thermal impact on peptide bioactivity during the spray drying process, which usually cannot be avoided but should be minimized. The co-drying of peptide with bulking agent (not necessarily encapsulation in this particular case), such as maltodextrin or starch, may help to protect peptide from severe denaturation [19,38,49]. Otherwise, lowering the inlet temperature could also help to protect the thermalsensitive material. Our previous research showed that the addition of ethanol could decrease the inlet temperature by 20 °C while maintain the same drying efficiency and powder quality, and lead to secondary structure change of protein, as shown in Fig. 2 [70].
4.2. Encapsulation and targeted release While microencapsulation is perhaps the area that has witnessed the greatest progress in the past few years for food peptide drying, there is still the need to optimise encapsulation efficiency and release profile, which play a crucial role in protecting the functionality of peptide [23,34,44,71]. As shown in Fig. 3, our previous work on the encapsulation of curcumin has illustrate the possibility using whey protein isolate as a wall material to increase the loading efficiency of curcumin by increasing their aqueous solubility through hydrophobic interaction with whey protein isolate [60]. In the future, strategies will be needed to increase encapsulation efficiency, lower wall material concentration, and introduce targeted release profile [70]. In particular, there is a need to develop special assays for encapsulated peptides to properly analyse the bioactivity & bioavailability of peptide after the encapsulation and release process [72,73].
Fig. 2. Ethanol as spray drying aids helps to lower the drying temperature and modify the structure of dried powders [67]. (A) Inlet temperature lowered from 200 °C to 180 °C when 30% ethanol was used; (B) Secondary structure of whey protein isolate (WPI) changed when different amount of ethanol was added.
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Fig. 3. SEM images of microparticles of whey protein isolate (WPI)-curcumin complexes spray dried at different inlet temperatures of (A) 150 °C and (B) 110 °C [58].
4.3. Granulation The granulation of food peptide powder may improve solubility in aqueous system and avoid fine particles when mixing with other food ingredients. In recent years, there have been significant advances in our understanding of how wetting, nucleation, growth and break-up combine to control granulation processes [74]. However, controlling granulation still remains a challenge for food peptide, with fundamental understanding of agglomeration and break-up still needed. As peptide/hydrolysate consist of smaller molecules than proteins and increasingly smaller powders are produced after spray drying, effective granulation is needed to reduce dust formation.
4.4. Masking flavour The need for new manufacturing technologies will be exacerbated in situations where peptide/hydrolysate powders may introduce unpleasant flavours into food matrix. Different combinations of spray drying parameters and encapsulation carriers may lead to some improvements, but it is difficult to completely solve the problem [75]. More recently, increasing interest has been shown in applying masking agents (such as xylitol, sucrose and maltodextrin) to improve the sensory properties [76].
5. Conclusion Food peptide/hydrolysate has various health benefits but the lack of suitable processing methods limits their range of applications in the food sector. We propose a set of parameters to be considered to optimise the spray drying of peptide powders. The focus on the following areas is needed to further improve the properties of spray-dried peptide powder, including thermal protection during drying, better encapsulation and targeted release, improved powder granulation, masking unpleasant taste or odour, extending shelf life, and enhancing compatibility in food matrix to further expand the application of peptide in functional foods.
Acknowledgement We acknowledge the funding for Yong Wang from the Department of Chemical Engineering, Monash University and for Cordelia Selomulya from the Australian Research Council (ARC) Future Fellowship (FT140101256).
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Perspective article
Spray drying strategy for encapsulation of bioactive peptide powders for food applications q Yong Wang a, Cordelia Selomulya a,b,⇑ a b
Department of Chemical Engineering, Monash University, 18 Alliance Lane, Clayton, VIC 3800, Australia School of Chemical Engineering, UNSW, Sydney, NSW 2052, Australia
a r t i c l e
i n f o
Article history: Received 1 July 2019 Received in revised form 28 October 2019 Accepted 30 October 2019 Available online xxxx Keywords: Bioactive peptide Spray drying Encapsulation Food proteins
a b s t r a c t Spray drying is commonly used to manufacture food and dairy powders at industrial scale. Bioactive peptides, as hydrolysis products from various food proteins, are gaining interest due to their biological functionality including immunomodulation, antimicrobial, and antioxidant properties. This article reviews several bioactive peptides/hydrolysates from different food sources, focusing on their processing and final powder characteristics. Additionally, we propose a strategy for encapsulation of food peptide/hydrolysate via spray drying. The identification of suitable drying parameters and/or formulation could help overcome the limitations associated with the current processing of food peptides. Ó 2019 The Society of Powder Technology Japan. Published by Elsevier B.V. and The Society of Powder Technology Japan. All rights reserved.
1. Introduction The application of peptides and protein hydrolysate to food is at the forefront of innovation for modern nutrition and represents a promising approach for efficient delivery of bioactive compounds. In line with this, various peptides for food applications have recently attracted interest. Bioactive peptides are specific protein fragments that are inactive within the sequence of the parent protein, but may exert various physiological functions after release during digestion [1]. In general, peptide refers to a more uniform protein fragment, while hydrolysate is a complex proteolytic mixture. However, unlike in the pharmaceutical industry, food peptide or protein hydrolysate do not have a strict distinction. Therefore, both terms will be used here interchangeably. Biological properties reported for peptides derived from food proteins include anti-aging, anti-cancer, anti-cariogenic, antidiabetic, anti-hypertensive, anti-microbial, anti-oxidative stress, anti-inflammatory, cholesterol lowering, growth enhancing, immunomodulatory, mineral binding, radical scavenging, regulation of glucose and insulin homeostasis, and satiety regulating [1–5]. In addition, the application of peptides into food is beneficial for populations with special nutritional needs such as infants, q This is the perspective article submitted for Virtual Special Issue: the 30th Anniversary Collection of Advanced Powder Technology. ⇑ Corresponding author at: Department of Chemical Engineering, Monash University, 18 Alliance Lane, Clayton, VIC 3800, Australia. E-mail addresses: [email protected], cordelia.selomulya@unsw. edu.au (C. Selomulya).
athletes and the elderly, due to its ease of digestion [6]. Thus the development of functional foods or nutraceuticals fortified with peptide is increasing [7]. Generally, food peptides require spray drying to be converted into powder for further applications. High cost of production, poor solubility of purified peptides powders, and consumer acceptance are among some of the challenges for commercialization of foodderived bioactive peptides [1,8]. Thus, a systematic design of spray drying trials is needed to optimize the process parameters to preserve the desirable attributes of the peptide. The effect of drying on the functional properties such as bioactivity and bioavailability of peptides needs to be considered. It is also necessary to account for the state of peptide particles after drying, including moisture content, shelf life, solubility, ease of use, taste/odour control, and others [9]. Since spray drying is one of the most energy consuming and expensive steps in the production of protein hydrolysates, care must also be taken to balance the energy consumption of the drying process, inlet/outlet temperature selection, and the final quality of the product. [10]. In industrial production, often a compromise must be made between the yield of hydrolysate that can be achieved versus the amount of water that must be removed to obtain a dry final product, e.g. adding less water during hydrolysis to minimize drying cost [10,11]. There are a number of studies on peptide that mainly focus on the amino acid composition [12–14], while microencapsulation of food peptide is also of interest [15]. Although with the same amino acid composition, the peptide functionality may vary greatly depending on the powder properties. Peptide is more fragmented
https://doi.org/10.1016/j.apt.2019.10.034 0921-8831/Ó 2019 The Society of Powder Technology Japan. Published by Elsevier B.V. and The Society of Powder Technology Japan. All rights reserved.
Please cite this article as: Y. Wang and C. Selomulya, Spray drying strategy for encapsulation of bioactive peptide powders for food applications, Advanced Powder Technology, https://doi.org/10.1016/j.apt.2019.10.034
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than protein, which makes the drying process more complicated if carrier is added into formulation and might interact with the bioactive peptides [9]. Another key consideration in encapsulation of peptides via spray drying is how well these thermally labile materials resist heat denaturation by hot air [16]. Although it is a common procedure to process protein-based food into powders, to the best of our knowledge, only limited studies have been done on developing a spray drying strategy to maximize bioactivity and functionality of food peptides, especially in their pure or concentrated forms. Here, we briefly summarized recent studies on the spray drying of food peptides, and propose areas for further investigation to achieve better quality and expand the application of food peptide powders. 2. Current research on food peptide drying The existing literature on food protein peptide or hydrolysate drying can be divided into three categories: (i) Direct drying of pure peptide [17,18]; (ii) Co-drying with processing aids (e.g. maltodextrin or starch), but not necessarily encapsulated even with claims of microencapsulation in some cases [19–21]; (iii) Microencapsulation (mainly using wall materials to encapsulate peptide, but may also use peptide as wall material to encapsulate other components) [9,22,23]. The main purpose of the first two categories is to manufacture powders rich in peptide for use as food or nutraceutical ingredients, while the main purpose of last category is on maintaining bioactivity or masking the smell/taste of the peptide. 2.1. Dairy peptide As major components of dairy protein, whey and casein peptides are spray dried and sold as powdered ingredients. Ma et al. compared the effects of spray drying and freeze drying on bioactivity, taste, and hygroscopicity of whey hydrolysate, and found that spray-dried pure whey hydrolysate showed similar bitter taste, hygroscopicity, and solubility compared to freeze-dried whey hydrolysate [17]. Contreras et al. compared the spray-dried and freeze-dried casein peptides, and found that the angiotensin converting enzyme (ACE)-inhibitory activity and the antihypertensive properties of casein hydrolysed products were maintained after spray drying or freeze drying [24]. Moreover, the spray-dried casein hydrolysate powder still showed their antihypertensive effects when added into yoghurts, indicating their potential as food ingredient [24]. The allergic properties of bovine milk proteins could be reduced by enzymatic hydrolysis [25]. But their relatively high cost and bitter taste may restrict the use of milk peptide in infant milk formula. However, recent studies demonstrated the stability of spray-dried whey hydrolysate powders throughout thermal and spray drying processes during infant milk formula manufacture, with the potential to lower drying cost by decreasing liquid viscosity, and increasing storage stability by reducing lactose crystallization [26,27]. On the other hand, the high hygroscopicity of whey hydrolysate powders may decrease the storage stability of infant milk formula by lowering the glass transition temperature during storage [28]. In addition, spray-dried whey protein hydrolysate were more susceptible to sticking due to the low glass transition temperature, also with lower surface polarity when compared to intact proteins, which need to be taken into consideration when using whey hydrolysate to fortify other dairy products [29,30]. Drusch et al. found that both casein hydrolysate and whey hydrolysate showed higher surface activity than casein or whey protein at the air-water interface, because of lower molecular weight and more ionisable groups. Thus the hydrolysed milk
proteins can be used to alter the surface composition of the spray-dried particles [31]. Better encapsulation of fish oil from oxidation was shown with whey hydrolysate as wall material, compared to b-lactoglobulin, reportedly due to higher accessibility of antioxidant amino acids from wall material due to enzymatic hydrolysis [22]. Microencapsulation of whey hydrolysate with whey protein and sodium alginate did not change the immunomodulatory activity, but could lower their hygroscopicity and solubility, and provide bitterness-masking effect [17]. Maltodextrin-encapsulated whey protein hydrolysate was significantly less hygroscopic than without encapsulation [32], while microencapsulation also helped to improve gastrointestinal stability of the whey protein hydrolysate [23]. For casein peptide, the bitter flavour can be reduced by spray drying with different carriers including gelatine and soy protein isolate, gum Arabic, and maltodextrin [33–36]. However, the rate of dissolution of casein hydrolysate in water may decrease significantly due to the introducing of wall material in encapsulation process [34]. 2.2. Chicken peptide Chicken meat could be hydrolysed and spray-dried into hydrolysate powders. Higher inlet temperature in spray drying resulted in lower powder moisture and bulk density, as well as spherical shape of hydrolysate powder [20]. In addition, high inlet air temperature (170–200 °C) in spray drying may also bring advantages like higher antioxidant activity of chicken meat hydrolysate powder as evidenced in precious study [37]. On the contrary, lower drying temperature (120–160 °C) combined with low feed flow rate could minimize the difference between inlet and outlet temperature in spray drying, in order to achieve high product recovery for chicken peptides [37]. Maltodextrin and gum Arabic could be used as carrier to improve the drying performance of chicken meat peptide by reducing the stickiness and hygroscopicity of hydrolysate powders, and also increasing their glass transition temperature [20,38]. Although freeze dried chicken skin hydrolysate showed higher water solubility, emulsion stability, and water holding capacity than the spray-dried powders, spray drying is still preferable due to its high production efficiency [39]. 2.3. Fish peptide Fish meat, especially the by-product of fillet, could be spraydried into powder for application in food or as animal feed. Spray-dried Black Tilapia fish hydrolysate powders showed high in vitro digestibility (up to 92.1%) at both inlet temperature (150 °C or 180 °C), and lower drying inlet temperature (150 °C) was preferable to achieve higher protein efficiency ratio in animal feed [21]. In another study, an inlet temperature of 185 °C was used to dry the protein hydrolysate of different variety of fish including Mackerel, White Fish, Blue Whiting, and Sprat [18]. The bitter flavours of dried fish hydrolysate powders were found to be relatively mild compared with the hydrolysates from milk proteins, showing their potential to be added into food matrix. Spraydried Sauvage fish protein hydrolysate powder (inlet temperature 180 °C) showed better oil absorption capacity and emulsion activity when the hydrolysis was finished by alkali or acid instead of enzyme, but the enzyme-hydrolysed powder has higher water solubility than the alkali or acid hydrolysis powders [40]. Pretreatment with ethanol could slow down the non-enzymatic browning of spray-dried fish hydrolysate powders (inlet temperature 180 °C), and also mask the bitter taste [41]. It has been suggested that despite the extensive studies of the hydrolysis process, further studies on the drying and storing of fish hydrolysates are necessary for industrial application [11,18].
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2.4. Other food peptides There are several features that may limit the application of plant proteins, including low solubility. Research on plant protein hydrolysate has the potential to bring new applications into the food industry. Spray-dried soy protein hydrolysate powder showed significantly higher solubility than soy protein isolate around pH 4.5, which is the isoelectric point of soybean protein. Soy hydrolysate also showed higher foaming capacity and lower viscosity, while retaining gel-forming ability, which is favourable for application in meat product [42]. Wang et al. investigated the effects of inlet drying air temperature (140–200 °C), feed flow rate (3– 12 mL/min), and solids concentration (25–40%) on the properties of spray-dried soy protein peptide powders, and found an optimized spray drying condition for soy peptide (160 °C inlet temperature, 6 mL/min feed rate, and 35% solid concentration) based on powder characteristics including colour, morphology, density, and antioxidant capacity [43]. Using maltodextrin as carrier, spray-dried flaxseed protein hydrolysate powder could not only show good solubility (92–96%), but also retain high antioxidant activity [44]. Egg white protein hydrolysate powder exhibited similar solubility and antioxidant activities when dried by either spray drying or freeze drying. However, spray drying could lower the emulsifying and foaming properties of egg white protein hydrolysate powder when compared with the freeze-dried powder, which might be caused by the low hydrophobicity of spray-dried peptide [45]. In another study, spray-dried egg white protein peptides (inlet temperature 180 °C) showed high in vitro angiotensin converting enzyme (ACE) inhibitory activity, which might be beneficial in preventing hypertension, heart failure, myocardial infarction, and diabetic nephropathy [46]. In particular, spray-dried egg white peptide Arg-Val-Pro-Ser-Leu (RVPSL) showed the potential as a functional food ingredient with highest ACE-inhibitory activity [47], while peptide Asp-His-Thr-Lys-Glu (DHTKE) had the best oxygen radical absorbance capacity [48]. Spray-dried mussel meat protein hydrolysate powders had a higher storage stability in comparison to chicken breast hydrolysate powder [49]. The addition of maltodextrin or gum Arabic as carrier might increase the glass transition temperature of mussel meat protein hydrolysate powders (from 64 °C to 69–97 °C) and decrease water adsorption, which is beneficial for storage stability [49]. The spray-dried mussel meat protein hydrolysate with maltodextrin carrier showed potential to be used as umami seasoning in instant noodles [50].
3. Drying strategy for food peptide Based on the analysis of existing data and our previous drying research experience, we recommend the following parameters to be considered when spray drying food protein peptide/hydrolysate into powders (shown in Fig. 1). Before drying, the hydrolysis process and the characteristics of the peptide/hydrolysate need to be considered. Several factors in the hydrolysis process affect the properties of hydrolysate, which in turn affects the drying process. The degree of hydrolysis, molecular weight distribution of the hydrolysate, and the amino acid composition may affect or even to some extent determined the water-binding ability of the peptide molecules [51]. This further affects the rheological property, surface charge, heat tolerance, and peptide interaction of the suspension before drying [52]. Other factors are easier to control, such as the moisture content and pH of the suspension, but also have a large impact on the subsequent drying process. Due to the variation in the functionality of peptides, the separation and purification steps can affect their
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bioactivity [4,53], which is crucial when claiming functionality of the ingredients in food. During drying, various parameters need to be adjusted carefully to accommodate the drying requirements of the peptide. Optimum drying conditions and tailored matrix formulations are required to avoid severe heat damage leading to loss in bioactivity [54]. Similar to the spray drying of other powders, the critical parameters that can be controlled are inlet temperature, outlet temperature and feeding rate [16]. Some operating parameters are determined by the type of spray dryer, such as evaporation efficiency and air flow, such that suitable conditions need to be determined for a specific dryer (e.g. drying aids or carriers) where computational modelling may help [55]. Unlike the drying of pure peptide, more factors need to be taken into consideration for encapsulation, including the carrier type, ratio between peptide and carrier, and pre-treatments, etc [16,56,57]. In addition, the understanding of droplet drying kinetics via single droplet drying may provide fundamental understanding of changes that occur in the peptide suspension during drying. Our works have demonstrated the use of single droplet drying to provide insight into micro-structural changes during drying [58,59], in addition to drying kinetics data that can be used in modelling the drying process including water vapor, drying rate, shrinking, etc [54,60,61]. Novel spray drying methodology, such as nano spray drying, developed to enhance powder solubility of pharmaceutical peptides, might also help to preserve the functionality of food peptide by reducing the droplet size and the drying duration [62]. After drying, the analysis of the powder is also critical. The physical characteristics of peptide powder, including particle size distribution, moisture content, morphology, and others, are relevant to properties such as solubility and shelf stability. The formation of porous granules can improve solubility in water, while the physical structure of the powder are correlated with the digestibility and peptide release [35,63].The thermal properties of peptide powder, such as glass transition temperature, together with other physical properties, will help to quantify flowability, stickiness, and stability of the peptide powder [64]. Both the functionality and sensory properties of peptide powders determine their applications in food matrix. In addition to solubility, emulsifying and foaming capacities will also determine the application of peptide powder [65]. Water activity and storage stability not only affects the peptide powder itself, but also affects the whole formulated food [26,66,67]. In some cases, the peptide powder may induce bitter taste and undesirable odour [9]. This requires consideration of the appropriate drying condition to mask the negative effect to food matrix. On the contrary, there are some cases where the peptide powder may introduce pleasant taste like umami, especially from those generated from fish and seaweed [1,68]. Bioavailability needs to be assessed after drying to ensure the bioactive peptides can still function normally [9,69]. The last thing to note is the complex composition of food matrix, where a comprehensive analysis is needed since any change can bring unexpected effect on flavour or texture.
4. Opportunities for research on food peptide powders In the previous section, although we try to list most of the technical areas to be considered in the peptide drying, there are some areas worthy of further investigation. These include how to maintain the bioactivity of peptide during the drying process, how to optimize the drying parameters to generate peptide powders with desired properties, and how to render peptide powders more compatible with the food matrix.
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Fig. 1. An overview of relevant process parameters and properties in the spray drying of food protein peptide/hydrolysate.
4.1. Drying protection The fundamental issue is how to develop methods for drying food peptide/hydrolysate without introducing unexpected changes in relevant powder properties. At the same time, consideration should be made regarding the thermal impact on peptide bioactivity during the spray drying process, which usually cannot be avoided but should be minimized. The co-drying of peptide with bulking agent (not necessarily encapsulation in this particular case), such as maltodextrin or starch, may help to protect peptide from severe denaturation [19,38,49]. Otherwise, lowering the inlet temperature could also help to protect the thermalsensitive material. Our previous research showed that the addition of ethanol could decrease the inlet temperature by 20 °C while maintain the same drying efficiency and powder quality, and lead to secondary structure change of protein, as shown in Fig. 2 [70].
4.2. Encapsulation and targeted release While microencapsulation is perhaps the area that has witnessed the greatest progress in the past few years for food peptide drying, there is still the need to optimise encapsulation efficiency and release profile, which play a crucial role in protecting the functionality of peptide [23,34,44,71]. As shown in Fig. 3, our previous work on the encapsulation of curcumin has illustrate the possibility using whey protein isolate as a wall material to increase the loading efficiency of curcumin by increasing their aqueous solubility through hydrophobic interaction with whey protein isolate [60]. In the future, strategies will be needed to increase encapsulation efficiency, lower wall material concentration, and introduce targeted release profile [70]. In particular, there is a need to develop special assays for encapsulated peptides to properly analyse the bioactivity & bioavailability of peptide after the encapsulation and release process [72,73].
Fig. 2. Ethanol as spray drying aids helps to lower the drying temperature and modify the structure of dried powders [67]. (A) Inlet temperature lowered from 200 °C to 180 °C when 30% ethanol was used; (B) Secondary structure of whey protein isolate (WPI) changed when different amount of ethanol was added.
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Fig. 3. SEM images of microparticles of whey protein isolate (WPI)-curcumin complexes spray dried at different inlet temperatures of (A) 150 °C and (B) 110 °C [58].
4.3. Granulation The granulation of food peptide powder may improve solubility in aqueous system and avoid fine particles when mixing with other food ingredients. In recent years, there have been significant advances in our understanding of how wetting, nucleation, growth and break-up combine to control granulation processes [74]. However, controlling granulation still remains a challenge for food peptide, with fundamental understanding of agglomeration and break-up still needed. As peptide/hydrolysate consist of smaller molecules than proteins and increasingly smaller powders are produced after spray drying, effective granulation is needed to reduce dust formation.
4.4. Masking flavour The need for new manufacturing technologies will be exacerbated in situations where peptide/hydrolysate powders may introduce unpleasant flavours into food matrix. Different combinations of spray drying parameters and encapsulation carriers may lead to some improvements, but it is difficult to completely solve the problem [75]. More recently, increasing interest has been shown in applying masking agents (such as xylitol, sucrose and maltodextrin) to improve the sensory properties [76].
5. Conclusion Food peptide/hydrolysate has various health benefits but the lack of suitable processing methods limits their range of applications in the food sector. We propose a set of parameters to be considered to optimise the spray drying of peptide powders. The focus on the following areas is needed to further improve the properties of spray-dried peptide powder, including thermal protection during drying, better encapsulation and targeted release, improved powder granulation, masking unpleasant taste or odour, extending shelf life, and enhancing compatibility in food matrix to further expand the application of peptide in functional foods.
Acknowledgement We acknowledge the funding for Yong Wang from the Department of Chemical Engineering, Monash University and for Cordelia Selomulya from the Australian Research Council (ARC) Future Fellowship (FT140101256).
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