- Email: [email protected]
Amino acids microbial production
S24
Abstracts / Journal of Biotechnology 185S (2014) S18–S36
ite Design (FCCCD) under Response Surface Methodology (RSM) using three important parameters, the optimal preparation condition of CLEA-lipase shows that the highest activity achieved is 9.407 U or 83% of the activity of the free lipase. It was prepared using 20% saturated (NH4 )2 SO4 as the precipitant, 60 mM glutaraldehyde as the cross-linker and 0.169 mM BSA as the feeder. The optimal reaction temperature and pH for both CLEA-lipase and free lipase differed, where they were 60 ◦ C and 8.2 and 45 ◦ C and 8 respectively. A systematic study of temperature and pH stability showed that CLEA-lipase is more stable than free lipase. Results also show that the prepared CLEA-lipase retained more than 50% of the initial activity after five repeated runs. The observed high stability and recyclability of CLEA-lipase prepared from CPH demonstrated that it has potential to be used in different industrial applications. http://dx.doi.org/10.1016/j.jbiotec.2014.07.079 Engineering of membrane desaturases with new specificities Pichová Iva ∗ , Bucek Aleˇs, Matouˇsková Petra, Svatoˇs Aleˇs Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Czech Republic E-mail address: [email protected] (P. Iva). Membrane-bound fatty acid desaturases (FADs) play a prominent role in the biosynthesis of unsaturated fatty acids (FA). The large variety of FA unsaturation patterns is enabled by the high diversity of substrate, regio-, and stereo-specificities of FADs but the lack of membrane desaturase crystal structures has made it difficult to elucidate the mechanism and control of these specificities. We identified novel insect desaturases that share high amino acid sequence identity but displays different regio- and stereospecificities. We prepared chimeras and point mutants of FADs from Manduca sexta and analysed their products by GC–MS. The results allowed us to identify the structural motif determining the desaturase specificities and to obtain desaturases with altered specificities. The results also revealed a flexibility of desaturases in accommodation of different FAs. Our findings represent a step toward understanding the mechanism of desaturation of fatty acids by membrane-bound desaturases and toward engineering of membrane desaturases with new specificities. This work has been supported by project TAO1011461 from TACR.
tion was observed from only olive waste. Apple and pomegranate wastes could not be utilized by the selected yeast. Pigment and biomass formation kinetics were performed during ten days under optimum growth conditions (22 ◦ C, 4,5 pH, 70% moisture) of the yeast. Astaxanthin was extracted chemically and determined by spectral analysis at its maximum wavelength value, 474 nm. 5.72–8.67 mg/L astaxanthin was produced from olive waste during the fermentation period. http://dx.doi.org/10.1016/j.jbiotec.2014.07.081 Novel expression strategies of enzymes for usage in food technology Sebastian Christopher Spohner 1,∗ , Hendrich Quitmann 2 , Peter Czermak 3 1 Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Giessen, Germany 2 Faculty of Biology and Chemistry, Justus Liebig University Giessen, Giessen, Germany 3 Department of Chemical Engineering, Kansas State University, Manhattan, USA
E-mail address: [email protected] (S.C. Spohner). Steviol glycosides are natural sweet tasting constituents of Stevia rebaudiana. This plant is native to subtropical and tropical regions of western North America to South America. The main steviol glycoside in the leaves is Stevioside with an amount of 5–10%. Further sweet compounds are Rebaudioside A (2–4%), Rebaudioside C (1–2%) and Dulcosid A (0.2–0.7%). Purified steviol glycosides are a white to slightly yellow powder. In December 2011, steviol glycosides were approved for sale in Europe as sweeteners (E960). Even highly purified steviol glycosides still possess taste attributes such as bitterness, liquorice flavour and sweet aftertaste. The flavour notes become more prominent as the concentration increases. For the enzymatic preparation of steviol glycosides enzymes can be used. Therefore specific fungal enzymes have been overexpressed in the methylotrophic yeast Pichia pastoris under the control of the constitutive GAP promoter using a novel innovative fast-forward high throughput cloning strategy. High amounts of enzyme have been harvested and concentrated via a simple ultrafiltration using ceramic membranes. The next steps will be the enzyme characterisation and a process development. http://dx.doi.org/10.1016/j.jbiotec.2014.07.082
http://dx.doi.org/10.1016/j.jbiotec.2014.07.080 Amino acids microbial production Food & Feed Biotechnology Astaxanthin pigment production by ATTC 24202 from fruit wastes using solid state fermentation method Derya Dursun ∗ , Efe Aytu˘g Altınc¸ic¸ek, Michael Bo˘guso˘glu, Ali Cos¸kun Dalgıc¸ Department of Food Engineering, Gaziantep University, Gaziantep, Turkey E-mail address: [email protected] (D. Dursun). Solid state fermentation (SSF) for the production of astaxanthin pigment by a specific yeast, ATCC 24202 (Xanthophyllomyces dendrorhous) from olive, apple and pomegranate wastes processed industrially was investigated in this study. Astaxanthin produc-
Alexander Borisiv Ratkov ∗ , Ignat Iankov Dimov, Rositza Plamenova Petkova, Julia Georgieva Krasteva, Filip Ivanov Filipov Department of Biotechnology, IM-BAS, Sofia, Bulgaria E-mail address: [email protected] (A.B. Ratkov). The presentation gives a short overview of the world market for amino acids. Attempt and achievements in investigations and developments of the technologies for some amino acids production at the Institute of Microbiology, Bulgarian Academy of Sciences are summarized. Improvements in selection of different microbial strain-producers for microbial production of l-lysine, l-valine, l-leucine as well as for simultaneous production of l-lysine plus lthreonine are briefly analyzed. Developments and achievements
Abstracts / Journal of Biotechnology 185S (2014) S18–S36
in bioprocess technology, i.e. development of lab scale, semiindustrial scale and production scale technologies for production of some of the above mentioned amino acids are summarized too. Specificity and efficiency of applying of fed-batch and repeated fedbatch (fed-batch with droppings) methods for cultivation of lysine producers as well as the industrial attempt of applying and regular production of the technology for microbial production of l-lysine from one side and for cultivation of the producers and technology specificities for production of branched chain amino acid (l-valine and l-leucine) from the other side are shortly discussed. http://dx.doi.org/10.1016/j.jbiotec.2014.07.083 Innovative approach for active food packaging using cinnamaldehyde Alfonsina Sara Tarantino 1 , Vincenzo Maria De Benedictis 2,∗ , Marta Madaghiele 1 , Christian Demitri 1 , Alessandro Sannino 1 1 University of Salento, Department of Innovation for Engineering, Via Monteroni, Campus Ecotekne, 73100 Lecce, Italy 2 High Tech District – DHITECH Scarl, via S. Trinchese, 61, 73100 Lecce, Italy
E-mail address: [email protected] (V.M. De Benedictis). In order to maintain food quality and freshness, it is necessary to select correct materials and packaging technologies. In this context, current tendencies include the development of packaging materials that interact with the product. Recently, increasing attention has been paid to develop and test films with antimicrobial properties that show promise in improving food safety and shelf life. The ultimate aim of this work is to develop a novel approach to improve food safety, by controlling the growth of food-borne and food-spoilage microorganisms, while reducing the use of synthetic preservatives. Bioactive films, based on chitosan and cinnamaldehyde, are envisaged for food packaging. Since the formation of the Schiff base between chitosan and cinnamaldehyde is reversible under suitable conditions, the films show the potential to activate the release of the antimicrobial cinnamaldehyde when necessary, and to control the release rate, thereby exerting either lethal or inhibitory effects against food pathogens or spoilage microorganisms present in foodstuffs. The studies demonstrate that small concentrations of CA are sufficient to actively functionalize chitosan substrates. Moreover, active food packaging based on chitosan films incorporating CA molecules can increase the shelf life of the product. http://dx.doi.org/10.1016/j.jbiotec.2014.07.084
S25
Catalase positive cocci and volatile compounds in fermented sausages Aybike Kamilo˘glu 1,∗ , Bilge Sayın 2 , Güzin Kaban 3 , Mükerrem Kaya 3 1 Department of Food Engineering, Bayburt University, Bayburt, Turkey 2 Department of Food Engineering, Ardahan University, Ardahan, Turkey 3 Department of Food Engineering, Ataturk University, Erzurum, Turkey
E-mail address: [email protected] (A. Kamilo˘glu). Aroma is a very important characteristic for fermented sausages. Aroma compounds are generated as a result of carbohydrate, protein and lipid degradation. In fermented sausages major volatiles of aroma components are developed from microbial degradation of aminoacids. Aromatic aminoacids (AAC) tyrosine, tryptophan and phenylalanine, branched-chain amino acids (BCAA) valine, leucine, isoleucine and sulphur containing amino acids (methionine and cysteine) are important amino acids for aroma formation in fermented sausages. Staphylococcus carnosus and Staphylococcus xylosus are preferred starter cultures for fermented sausages. They have several effects on technological characteristics such as catalase activity, colour stability, nitrite and nitrate reduction and decreasing rancid odour by preventing oxidation of fatty acids. Also, their proteolytic and lipolytic activities enhance the quality of fermented sausages by generation of flavour-active compounds and their precursors, such as amino acids, free fatty acids, aldehydes, amines, ketons, and alcohols. In this review, effects of catalase positive cocci, especially S. carnosus and S. xylosus, on the flavour of fermented sausage are discussed. http://dx.doi.org/10.1016/j.jbiotec.2014.07.085 Probiotic meat products Bilge Sayın 1,∗ , Aybike Kamilo˘glu 2 , Mükerrem Kaya 3 , Güzin Kaban 3 1 Department of Food Engineering, Ardahan University, Ardahan, Turkey 2 Department of Food Engineering, Bayburt University, Bayburt, Turkey 3 Department of Food Engineering, Atatürk University, Erzurum, Turkey
E-mail address: [email protected] (B. Sayın). Fermentation is probably one of the oldest forms of food preservation. Fermentation is an important phase in the production of many food products all over the world. In recent years, there has been a growing acceptance on that fermented meat products are important mediums in transferring probiotic microorganisms to human body. Probiotic bacteria that can be used in the manufacturing of dry fermented sausages should survive in fermented products. Also, these microorganisms should dominate other microorganisms found in finished products. On the other hand, it is an important factor that bacterial viability may be affected by the high curing salt and the low values of water activity and pH in the products. Therefore, characteristics of probiotic bacteria strains used in the production play important roles. Thus, strains demonstrating both probiotic potential and technological performance would bring about advantages for dry fermented sausages. These strains should have such abilities as the stomach acidity resistance, lysozyme and bile resistance and the ability to colonize the human intestinal tract using mechanisms of adhesion or binding to intesti-
Abstracts / Journal of Biotechnology 185S (2014) S18–S36
ite Design (FCCCD) under Response Surface Methodology (RSM) using three important parameters, the optimal preparation condition of CLEA-lipase shows that the highest activity achieved is 9.407 U or 83% of the activity of the free lipase. It was prepared using 20% saturated (NH4 )2 SO4 as the precipitant, 60 mM glutaraldehyde as the cross-linker and 0.169 mM BSA as the feeder. The optimal reaction temperature and pH for both CLEA-lipase and free lipase differed, where they were 60 ◦ C and 8.2 and 45 ◦ C and 8 respectively. A systematic study of temperature and pH stability showed that CLEA-lipase is more stable than free lipase. Results also show that the prepared CLEA-lipase retained more than 50% of the initial activity after five repeated runs. The observed high stability and recyclability of CLEA-lipase prepared from CPH demonstrated that it has potential to be used in different industrial applications. http://dx.doi.org/10.1016/j.jbiotec.2014.07.079 Engineering of membrane desaturases with new specificities Pichová Iva ∗ , Bucek Aleˇs, Matouˇsková Petra, Svatoˇs Aleˇs Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Czech Republic E-mail address: [email protected] (P. Iva). Membrane-bound fatty acid desaturases (FADs) play a prominent role in the biosynthesis of unsaturated fatty acids (FA). The large variety of FA unsaturation patterns is enabled by the high diversity of substrate, regio-, and stereo-specificities of FADs but the lack of membrane desaturase crystal structures has made it difficult to elucidate the mechanism and control of these specificities. We identified novel insect desaturases that share high amino acid sequence identity but displays different regio- and stereospecificities. We prepared chimeras and point mutants of FADs from Manduca sexta and analysed their products by GC–MS. The results allowed us to identify the structural motif determining the desaturase specificities and to obtain desaturases with altered specificities. The results also revealed a flexibility of desaturases in accommodation of different FAs. Our findings represent a step toward understanding the mechanism of desaturation of fatty acids by membrane-bound desaturases and toward engineering of membrane desaturases with new specificities. This work has been supported by project TAO1011461 from TACR.
tion was observed from only olive waste. Apple and pomegranate wastes could not be utilized by the selected yeast. Pigment and biomass formation kinetics were performed during ten days under optimum growth conditions (22 ◦ C, 4,5 pH, 70% moisture) of the yeast. Astaxanthin was extracted chemically and determined by spectral analysis at its maximum wavelength value, 474 nm. 5.72–8.67 mg/L astaxanthin was produced from olive waste during the fermentation period. http://dx.doi.org/10.1016/j.jbiotec.2014.07.081 Novel expression strategies of enzymes for usage in food technology Sebastian Christopher Spohner 1,∗ , Hendrich Quitmann 2 , Peter Czermak 3 1 Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Giessen, Germany 2 Faculty of Biology and Chemistry, Justus Liebig University Giessen, Giessen, Germany 3 Department of Chemical Engineering, Kansas State University, Manhattan, USA
E-mail address: [email protected] (S.C. Spohner). Steviol glycosides are natural sweet tasting constituents of Stevia rebaudiana. This plant is native to subtropical and tropical regions of western North America to South America. The main steviol glycoside in the leaves is Stevioside with an amount of 5–10%. Further sweet compounds are Rebaudioside A (2–4%), Rebaudioside C (1–2%) and Dulcosid A (0.2–0.7%). Purified steviol glycosides are a white to slightly yellow powder. In December 2011, steviol glycosides were approved for sale in Europe as sweeteners (E960). Even highly purified steviol glycosides still possess taste attributes such as bitterness, liquorice flavour and sweet aftertaste. The flavour notes become more prominent as the concentration increases. For the enzymatic preparation of steviol glycosides enzymes can be used. Therefore specific fungal enzymes have been overexpressed in the methylotrophic yeast Pichia pastoris under the control of the constitutive GAP promoter using a novel innovative fast-forward high throughput cloning strategy. High amounts of enzyme have been harvested and concentrated via a simple ultrafiltration using ceramic membranes. The next steps will be the enzyme characterisation and a process development. http://dx.doi.org/10.1016/j.jbiotec.2014.07.082
http://dx.doi.org/10.1016/j.jbiotec.2014.07.080 Amino acids microbial production Food & Feed Biotechnology Astaxanthin pigment production by ATTC 24202 from fruit wastes using solid state fermentation method Derya Dursun ∗ , Efe Aytu˘g Altınc¸ic¸ek, Michael Bo˘guso˘glu, Ali Cos¸kun Dalgıc¸ Department of Food Engineering, Gaziantep University, Gaziantep, Turkey E-mail address: [email protected] (D. Dursun). Solid state fermentation (SSF) for the production of astaxanthin pigment by a specific yeast, ATCC 24202 (Xanthophyllomyces dendrorhous) from olive, apple and pomegranate wastes processed industrially was investigated in this study. Astaxanthin produc-
Alexander Borisiv Ratkov ∗ , Ignat Iankov Dimov, Rositza Plamenova Petkova, Julia Georgieva Krasteva, Filip Ivanov Filipov Department of Biotechnology, IM-BAS, Sofia, Bulgaria E-mail address: [email protected] (A.B. Ratkov). The presentation gives a short overview of the world market for amino acids. Attempt and achievements in investigations and developments of the technologies for some amino acids production at the Institute of Microbiology, Bulgarian Academy of Sciences are summarized. Improvements in selection of different microbial strain-producers for microbial production of l-lysine, l-valine, l-leucine as well as for simultaneous production of l-lysine plus lthreonine are briefly analyzed. Developments and achievements
Abstracts / Journal of Biotechnology 185S (2014) S18–S36
in bioprocess technology, i.e. development of lab scale, semiindustrial scale and production scale technologies for production of some of the above mentioned amino acids are summarized too. Specificity and efficiency of applying of fed-batch and repeated fedbatch (fed-batch with droppings) methods for cultivation of lysine producers as well as the industrial attempt of applying and regular production of the technology for microbial production of l-lysine from one side and for cultivation of the producers and technology specificities for production of branched chain amino acid (l-valine and l-leucine) from the other side are shortly discussed. http://dx.doi.org/10.1016/j.jbiotec.2014.07.083 Innovative approach for active food packaging using cinnamaldehyde Alfonsina Sara Tarantino 1 , Vincenzo Maria De Benedictis 2,∗ , Marta Madaghiele 1 , Christian Demitri 1 , Alessandro Sannino 1 1 University of Salento, Department of Innovation for Engineering, Via Monteroni, Campus Ecotekne, 73100 Lecce, Italy 2 High Tech District – DHITECH Scarl, via S. Trinchese, 61, 73100 Lecce, Italy
E-mail address: [email protected] (V.M. De Benedictis). In order to maintain food quality and freshness, it is necessary to select correct materials and packaging technologies. In this context, current tendencies include the development of packaging materials that interact with the product. Recently, increasing attention has been paid to develop and test films with antimicrobial properties that show promise in improving food safety and shelf life. The ultimate aim of this work is to develop a novel approach to improve food safety, by controlling the growth of food-borne and food-spoilage microorganisms, while reducing the use of synthetic preservatives. Bioactive films, based on chitosan and cinnamaldehyde, are envisaged for food packaging. Since the formation of the Schiff base between chitosan and cinnamaldehyde is reversible under suitable conditions, the films show the potential to activate the release of the antimicrobial cinnamaldehyde when necessary, and to control the release rate, thereby exerting either lethal or inhibitory effects against food pathogens or spoilage microorganisms present in foodstuffs. The studies demonstrate that small concentrations of CA are sufficient to actively functionalize chitosan substrates. Moreover, active food packaging based on chitosan films incorporating CA molecules can increase the shelf life of the product. http://dx.doi.org/10.1016/j.jbiotec.2014.07.084
S25
Catalase positive cocci and volatile compounds in fermented sausages Aybike Kamilo˘glu 1,∗ , Bilge Sayın 2 , Güzin Kaban 3 , Mükerrem Kaya 3 1 Department of Food Engineering, Bayburt University, Bayburt, Turkey 2 Department of Food Engineering, Ardahan University, Ardahan, Turkey 3 Department of Food Engineering, Ataturk University, Erzurum, Turkey
E-mail address: [email protected] (A. Kamilo˘glu). Aroma is a very important characteristic for fermented sausages. Aroma compounds are generated as a result of carbohydrate, protein and lipid degradation. In fermented sausages major volatiles of aroma components are developed from microbial degradation of aminoacids. Aromatic aminoacids (AAC) tyrosine, tryptophan and phenylalanine, branched-chain amino acids (BCAA) valine, leucine, isoleucine and sulphur containing amino acids (methionine and cysteine) are important amino acids for aroma formation in fermented sausages. Staphylococcus carnosus and Staphylococcus xylosus are preferred starter cultures for fermented sausages. They have several effects on technological characteristics such as catalase activity, colour stability, nitrite and nitrate reduction and decreasing rancid odour by preventing oxidation of fatty acids. Also, their proteolytic and lipolytic activities enhance the quality of fermented sausages by generation of flavour-active compounds and their precursors, such as amino acids, free fatty acids, aldehydes, amines, ketons, and alcohols. In this review, effects of catalase positive cocci, especially S. carnosus and S. xylosus, on the flavour of fermented sausage are discussed. http://dx.doi.org/10.1016/j.jbiotec.2014.07.085 Probiotic meat products Bilge Sayın 1,∗ , Aybike Kamilo˘glu 2 , Mükerrem Kaya 3 , Güzin Kaban 3 1 Department of Food Engineering, Ardahan University, Ardahan, Turkey 2 Department of Food Engineering, Bayburt University, Bayburt, Turkey 3 Department of Food Engineering, Atatürk University, Erzurum, Turkey
E-mail address: [email protected] (B. Sayın). Fermentation is probably one of the oldest forms of food preservation. Fermentation is an important phase in the production of many food products all over the world. In recent years, there has been a growing acceptance on that fermented meat products are important mediums in transferring probiotic microorganisms to human body. Probiotic bacteria that can be used in the manufacturing of dry fermented sausages should survive in fermented products. Also, these microorganisms should dominate other microorganisms found in finished products. On the other hand, it is an important factor that bacterial viability may be affected by the high curing salt and the low values of water activity and pH in the products. Therefore, characteristics of probiotic bacteria strains used in the production play important roles. Thus, strains demonstrating both probiotic potential and technological performance would bring about advantages for dry fermented sausages. These strains should have such abilities as the stomach acidity resistance, lysozyme and bile resistance and the ability to colonize the human intestinal tract using mechanisms of adhesion or binding to intesti-