牦牛乳源嗜热链球菌直投式发酵剂的制备及应用
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摘要
为制备优良功能的牦牛乳直投式发酵剂,在脱脂乳添加量、甘油添加量、海藻糖添加量、谷氨酸钠添加量4个单因素基础上,选用正交试验优化菌株保护剂的工艺条件,并运用真空包装技术和2种国家食品检验标准、数字旋转黏度计以及专业人员的感官评价等方法,考察发酵剂的发酵性能。结果表明,菌株在p H 6. 86的缓冲剂、6 000 r/min离心10 min,以及-20℃的预冻条件下生长最佳;而脱脂乳、谷氨酸钠、甘油、海藻糖质量浓度分别为120、10、60和80 g/L的条件下,菌株存活率为90. 08%,发酵剂发酵性能最佳。该发酵剂应用于发酵酸奶生产中,酸奶具有活菌数高、发酵性能优、黏度高并具备改善牦牛乳的风味等感官优点。
In order to prepare an excellent yak milk vat starter,the amounts of skimmed milk,glycerin,trehalose and sodium glutamate added were used as single factors for orthogonal test to optimize the strain protectant. The starter was vacuum-packed and its fermentation condition was measured by two national food inspection standards,digital rotational viscometers and professional sensory evaluation. The results showed that the strain grew the best when centrifuged at 6 000 r/min for 10 min and pre-frozen at-20°C in a p H 6. 86 buffer. Moreover,the survival rate of the strain was 90. 08% and the fermentation performance of the starter was the best when adding 120 g/L skimmed milk,10 g/L sodium glutamate,60 g/L glycerin and 80 g/L trehalose. Furthermore,the yoghurt produced with this agent had characteristics of high viable bacteria,excellent fermentation performance,high viscosity,and improved flavor and texture of yak milk.
In order to prepare an excellent yak milk vat starter,the amounts of skimmed milk,glycerin,trehalose and sodium glutamate added were used as single factors for orthogonal test to optimize the strain protectant. The starter was vacuum-packed and its fermentation condition was measured by two national food inspection standards,digital rotational viscometers and professional sensory evaluation. The results showed that the strain grew the best when centrifuged at 6 000 r/min for 10 min and pre-frozen at-20°C in a p H 6. 86 buffer. Moreover,the survival rate of the strain was 90. 08% and the fermentation performance of the starter was the best when adding 120 g/L skimmed milk,10 g/L sodium glutamate,60 g/L glycerin and 80 g/L trehalose. Furthermore,the yoghurt produced with this agent had characteristics of high viable bacteria,excellent fermentation performance,high viscosity,and improved flavor and texture of yak milk.
引文
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[9]刘刚,梁琪,宋雪梅,等. Plackett-Burman和Box-Behnken试验优化嗜热链球菌Q4F8产胞外多糖工艺[J/OL].食品科学,2019:1-15.
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[11] DONG C W,MAO D B,BAI Y H,et al. Screening and identification of lactic acid bacterium producing bacteriocin with broad spectrum[J]. Science and Technology of Food Industry,2009(4):129-133.
[12] LI C,SUN,J W,QI X X,et al. Na Cl stress impact on the key enzymes in glycolysis from Lactobacillus bulgaricus during freeze-drying[J]. Brazilian Journal of Microbiology,2015,46(4):1 193-1 199.
[13] GARCA G B,ROMERO R A,VZQUEZ O L,et al.Sensory evaluation of low-fat yoghurt produced with microbial transglutaminase and comparison with physicochemical evaluation[J]. Journal of the Science of Food and Agriculture,2018,11(4):21-29.
[14] PANG X Y,ZHANG S W,LU J,et al. Identification and functional validation of autolysis—associated genes in Lactobacillus bulgaricus ATCC BAA-365[J]. Frontiers in Microbiology,2017,8:86-97.
[15] ZIMMERMAN T,GYAWALI R,IBRAHIM S. Autolyse the cell in order to save it? Inducing,then blocking,autolysis as a strategy for delaying cell death in the probiotic Lactobacillus reuteri[J]. Biotechnology Letters,2017,39(10):1 547-1 551.
[16]宋雪梅,张炎,张卫兵,等.成熟温度对牦牛乳硬质干酪中乳酸菌自溶及氨肽酶活性的影响[J].食品与发酵工业,2016,42(5):38-43.
[17] FUGL A,BERHE T,KIRAN A,et al. Characterisation of lactic acid bacteria in spontaneously fermented camel milk and selection of strains for fermentation of camel milk[J]. International Dairy Journal,2017,73(2):19-24.
[18] FERNANDA S,GEORGES C,CATHERINE B. Effect of centrifugation conditions on the cryotolerance of Lactobacillus bulgaricus CFL1[J]. Food and Bioprocess Technology,2010,3(1):36-42.
[19] ERVA N,KARASU E. Determination of the effect of exopolysaccharide(EPS)from Lactobacillus brevis E25 on adhesion of food powders on the surfaces,using the centrifuge technique[J]. Journal of Food Engineering,2019,242(7):106-114.
[20] BRAVO B M,BRIZUELA N,GERBINO E,et al. Effect of protective agents and previous acclimation on ethanol resistance of frozen and freeze-dried Lactobacillus plantarum strains[J]. Cryobiology,2015,71(3):522-528.
[21] CHANG-HO K,YONG G K,SEUL H H,et al. Effect of pumpkin powder as cryoprotectant to improve the viability of freeze dried lactic acid bacteria[J]. KSBB Journal,2017,32(4):251-255.
[22] LU Y J,HUANG L J,YANG T B,et al. Optimization of a cryoprotective medium to increase the viability of freezedried Streptococcus thermophilus by response surface methodology[J]. LWT-Food Science and Technology,2017,80:92-97.
[23] MARAGKOUDAKIS P,MIARIS C,ROJEZ P,et al.Production of traditional Greek yoghurt using Lactobacillus strains with probiotic potential as starter adjuncts[J]. International Dairy Journal,2006,16(1):52-60.
[24] MENDOZA G M,PASTERIS S E. OTERO M C,et al.Survival and beneficial properties of lactic acid bacteria from raniculture subjected to freeze-drying and storage[J]. Journal of Applied Microbiology,2014,116(1):157-166.
[25] BONATSOU S,ILIOPOULOS V,MALLOUCHOS A,et al. Effect of osmotic dehydration of olives as pre-fermentation treatment and partial substitution of sodium chloride by monosodium glutamate in the fermentation profile of Kalamata natural black olives[J]. Food Microbiol,2017,63(1):72-83.
[26] SANTIVARANGKNA C,WEMXIING M,FOERST P,et al. Damage of cell envelope of Lactobacillus helveticus during vacuum drying[J]. Journal of Applied Microbiology,2007,102(3):748-756.
[27] Raquel F S,Amanda A R M,Carlos P C,et al. Trehalose as a cryoprotectant in freeze-dried wheat sourdough production[J]. LWT-Food Science and Technology,2018,89(3):510-517.
[28] CARVALHO A S,SILVA J,HO P,et al. Survival of freeze-dried Lactobacillus plantarum and Lactobacillus rhamnosus during storage in the presence of protectants[J]. Biotechnology Letters,2002,24(19):1 587-1 591.
[29] SYAMALADEVI R M,TANG J,VILLA-ROJAS R,et al. Influence of water activity on thermal resistance of microorganisms in low-moisture foods:A review[J]. Comprehensive Reviews in Food Science&Food Safety,2016,15(2):353-370.
[30] MIAO S,MILLS S,STANTON C,et al. Effect of disaccharides on survival during storage of freeze dried probiotics[J]. Science and Technology,2008,88(1):19-30.
[31] TEIXEIRA P,KIRBY R,CASTRO H. Evidence of membrane lipid oxidation of spray-dried Lactobacillus bulgaricus during storage[J]. Letters in Applied Microbiology,1996,22(1):34-38.
[32]张炎,张卫兵,宋雪梅,等.冻干保护剂对复合低温乳酸菌发酵剂的影响[J].食品工业科技,2015,36(7):144-147; 153.
[33] BUTTRISS J. Milk:Dietary importance[M]. Encyclopedia of Food Sciences and Nutrition,2003:3 968-3 974.
[34]秦虹,梁琪,张卫兵.甘肃藏区自然发酵耗牛乳中优良乳酸菌的筛选及鉴定[J].食品科学,2013,34(19):241-246.
[2]施奇静,王静,孙俊平,等.中国牦牛资源保护及可持续利用:驯化与品种培育[J].家畜生态学报,2016,37(1):81-86.
[3] VELLY H,BOUIX M,PASSOT S,et al. Cyclopropanation of unsaturated fatty acids and membrane rigidifcation improve the freezedrying resistance of Lactococcus lactis subsp. lactis TOMSC161[J]. Applied Microbiology and Biotechnology,2015,99(2):907-918.
[4] PEIREN J,BUYSE J,DE VOS P,et al. Improving survival and storage stability of bacteria recalcitrant to freezedrying:a coordinated study by European culture collections[J]. Applied Microbiology and Biotechnology,2015,99(8):3 559-3 571.
[5] CHEN H,CHEN S W,LI C N,et al. Response surface optimization of lyoprotectant for Lactobacillus bulgaricus during vacuum freezedrying[J]. Preparative Biochemistry and Biotechnology,2015,45(5):463-475.
[6] ROSEN R,BUCHINGER S,PFNDER R,et al. SOS gene induction and possible mutagenic effects of freeze-drying in Escherichia coli and Salmonella typhimurium[J].Applied Microbiology and Biotechnology,2016,100(21):9 255-9 264.
[7] BASHOLLI-SALIHU M,MUELLER M,SALAR-BEHZADI S. Effect of lyoprotectants onβ-glucosidase activity and viability of Bifidobacterium infantis after freeze-drying and storage in milk and low p H juices[J]. LWT-Food Science and Technology,2014,57(1):276-282.
[8] NAHR F K,MOKARRAM R R,HEJAZI M A,et al. Optimization of the nanocellulose based cryoprotective medium to enhance the viability of freeze dried Lactobacillus plantarum using response surface methodology[J]. LWT-Food Science and Technology,2015,64(1):326-332.
[9]刘刚,梁琪,宋雪梅,等. Plackett-Burman和Box-Behnken试验优化嗜热链球菌Q4F8产胞外多糖工艺[J/OL].食品科学,2019:1-15.
[10]郭兴华,凌代文.乳酸细菌现代研究实验技术[M].北京:科学出版社,2013:69-70.
[11] DONG C W,MAO D B,BAI Y H,et al. Screening and identification of lactic acid bacterium producing bacteriocin with broad spectrum[J]. Science and Technology of Food Industry,2009(4):129-133.
[12] LI C,SUN,J W,QI X X,et al. Na Cl stress impact on the key enzymes in glycolysis from Lactobacillus bulgaricus during freeze-drying[J]. Brazilian Journal of Microbiology,2015,46(4):1 193-1 199.
[13] GARCA G B,ROMERO R A,VZQUEZ O L,et al.Sensory evaluation of low-fat yoghurt produced with microbial transglutaminase and comparison with physicochemical evaluation[J]. Journal of the Science of Food and Agriculture,2018,11(4):21-29.
[14] PANG X Y,ZHANG S W,LU J,et al. Identification and functional validation of autolysis—associated genes in Lactobacillus bulgaricus ATCC BAA-365[J]. Frontiers in Microbiology,2017,8:86-97.
[15] ZIMMERMAN T,GYAWALI R,IBRAHIM S. Autolyse the cell in order to save it? Inducing,then blocking,autolysis as a strategy for delaying cell death in the probiotic Lactobacillus reuteri[J]. Biotechnology Letters,2017,39(10):1 547-1 551.
[16]宋雪梅,张炎,张卫兵,等.成熟温度对牦牛乳硬质干酪中乳酸菌自溶及氨肽酶活性的影响[J].食品与发酵工业,2016,42(5):38-43.
[17] FUGL A,BERHE T,KIRAN A,et al. Characterisation of lactic acid bacteria in spontaneously fermented camel milk and selection of strains for fermentation of camel milk[J]. International Dairy Journal,2017,73(2):19-24.
[18] FERNANDA S,GEORGES C,CATHERINE B. Effect of centrifugation conditions on the cryotolerance of Lactobacillus bulgaricus CFL1[J]. Food and Bioprocess Technology,2010,3(1):36-42.
[19] ERVA N,KARASU E. Determination of the effect of exopolysaccharide(EPS)from Lactobacillus brevis E25 on adhesion of food powders on the surfaces,using the centrifuge technique[J]. Journal of Food Engineering,2019,242(7):106-114.
[20] BRAVO B M,BRIZUELA N,GERBINO E,et al. Effect of protective agents and previous acclimation on ethanol resistance of frozen and freeze-dried Lactobacillus plantarum strains[J]. Cryobiology,2015,71(3):522-528.
[21] CHANG-HO K,YONG G K,SEUL H H,et al. Effect of pumpkin powder as cryoprotectant to improve the viability of freeze dried lactic acid bacteria[J]. KSBB Journal,2017,32(4):251-255.
[22] LU Y J,HUANG L J,YANG T B,et al. Optimization of a cryoprotective medium to increase the viability of freezedried Streptococcus thermophilus by response surface methodology[J]. LWT-Food Science and Technology,2017,80:92-97.
[23] MARAGKOUDAKIS P,MIARIS C,ROJEZ P,et al.Production of traditional Greek yoghurt using Lactobacillus strains with probiotic potential as starter adjuncts[J]. International Dairy Journal,2006,16(1):52-60.
[24] MENDOZA G M,PASTERIS S E. OTERO M C,et al.Survival and beneficial properties of lactic acid bacteria from raniculture subjected to freeze-drying and storage[J]. Journal of Applied Microbiology,2014,116(1):157-166.
[25] BONATSOU S,ILIOPOULOS V,MALLOUCHOS A,et al. Effect of osmotic dehydration of olives as pre-fermentation treatment and partial substitution of sodium chloride by monosodium glutamate in the fermentation profile of Kalamata natural black olives[J]. Food Microbiol,2017,63(1):72-83.
[26] SANTIVARANGKNA C,WEMXIING M,FOERST P,et al. Damage of cell envelope of Lactobacillus helveticus during vacuum drying[J]. Journal of Applied Microbiology,2007,102(3):748-756.
[27] Raquel F S,Amanda A R M,Carlos P C,et al. Trehalose as a cryoprotectant in freeze-dried wheat sourdough production[J]. LWT-Food Science and Technology,2018,89(3):510-517.
[28] CARVALHO A S,SILVA J,HO P,et al. Survival of freeze-dried Lactobacillus plantarum and Lactobacillus rhamnosus during storage in the presence of protectants[J]. Biotechnology Letters,2002,24(19):1 587-1 591.
[29] SYAMALADEVI R M,TANG J,VILLA-ROJAS R,et al. Influence of water activity on thermal resistance of microorganisms in low-moisture foods:A review[J]. Comprehensive Reviews in Food Science&Food Safety,2016,15(2):353-370.
[30] MIAO S,MILLS S,STANTON C,et al. Effect of disaccharides on survival during storage of freeze dried probiotics[J]. Science and Technology,2008,88(1):19-30.
[31] TEIXEIRA P,KIRBY R,CASTRO H. Evidence of membrane lipid oxidation of spray-dried Lactobacillus bulgaricus during storage[J]. Letters in Applied Microbiology,1996,22(1):34-38.
[32]张炎,张卫兵,宋雪梅,等.冻干保护剂对复合低温乳酸菌发酵剂的影响[J].食品工业科技,2015,36(7):144-147; 153.
[33] BUTTRISS J. Milk:Dietary importance[M]. Encyclopedia of Food Sciences and Nutrition,2003:3 968-3 974.
[34]秦虹,梁琪,张卫兵.甘肃藏区自然发酵耗牛乳中优良乳酸菌的筛选及鉴定[J].食品科学,2013,34(19):241-246.
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