低温低酸发酵牛肉专用高效浓缩发酵剂的制备工艺研究
|
摘要
本课题以国产发酵牛肉Bundnerfleisch自然发酵过程中分离筛选出的三株优势菌——植物乳杆菌、戊糖片球菌和木糖葡萄球菌为研究对象,对菌株部分特性进行了测定,系统研究了菌体细胞增殖条件、浓缩分离条件、冻干菌粉发酵剂冷冻过程中抗冷冻复合干燥剂的配制、成品发酵剂的贮存特性,旨在为低酸发酵肉制品直投式浓缩型发酵剂生产提供依据。研究结果如下:
本文测定了植物乳杆菌、戊糖片球菌和木糖葡萄球菌的部分发酵特性,研究发现:三者均具有较强的耐盐性和耐亚硝酸盐性;植物乳杆菌和戊糖片球菌只具有蛋白降解能力,而木糖葡萄球菌具有蛋白降解能力和脂肪降解能力;植物乳杆菌和戊糖片球菌菌株对致病菌的抑菌效果较好,木糖葡萄球菌抑菌作用较差。
对植物乳杆菌、戊糖片球菌和木糖葡萄球菌的增殖培养基进行了优化,确定了菌株的复合增殖培养基配方。植物乳杆菌的复合增殖培养基配方为:麦芽汁培养基中添加2%乳糖、1%牛肉蛋白胨、2.5%酵母膏、10%番茄汁;戊糖片球菌的复合增殖培养基配方为:麦芽汁培养基中添加0.5%乳糖、1.5%牛肉蛋白胨、5%酵母膏、10%番茄汁;木糖葡萄球菌的复合增殖培养基配方为:麦芽汁培养基中添加0.5%葡萄糖、1.5%牛肉蛋白胨、0.1%牛肉膏、1%NaCl。
同时对菌株的增殖培养条件进行了研究,确定了菌株的培养条件。植物乳杆菌和戊糖片球菌:培养温度为20℃,培养基初始pH值6.5～7.0,培养时间20h,菌体密度可分别达3.75x10~9cfu/mL和3.68×10~9cfu/mL:木糖葡萄球菌:培养温度为20℃,培养基初始pH值7.0,培养时间20h,菌体密度可达5.00x10~9cfu/mL。
对菌体细胞浓缩分离条件进行了选择研究,结果显示:离心方法浓缩菌体的效果明显高于细菌过滤器的浓缩效果,并且采用高转速离心,可以提高浓缩菌体浓缩倍数,有利于菌体细胞富集分离。确定菌体细胞浓缩分离的适宜离心条件为0～4℃、6000rpm离心30min。
以12%的脱脂乳为冻干悬浮基质添加单因子制成冻干保护剂,进行冷冻干燥试验。从试验结果中发现蔗糖、维生素C、甘油和CaCO_3是植物乳杆菌、戊糖片球菌和木糖葡萄球菌的较好单因子保护剂。利用L_9(3~4)正交试验优化的植物乳杆菌、戊糖片球菌和木糖葡萄球菌的复合冻干保护剂配方分别为:在脱脂乳中添加5%蔗糖、0.5%维生素C、4%甘油、3%CACO_3;脱脂乳中添加4%蔗糖、1%维生素C、3%甘油、2%CACO_3和脱脂乳中添加3%蔗糖、1.5%维生素C、5%甘油、4%CACO_3,可分别使植物乳杆菌、戊糖片球菌和木糖葡萄球菌冷冻干燥存活率达到63.01%、66.20%和70.47%,制备的冻干菌粉发酵剂的活菌数分别为2.62×10~(10)cfu/g、2.54×10~(10)cfu/g和2.91×10~(10)cfu/g。
Biotechnological characteristics, high density cultivation, combined cryoprotectants of concentrated cultures production and quality guarantee-time of starts cultures.et al. were studied here systemically with three strains including Lactobacillus plantarum, Pediococcus pentosaceus and Staphylococcus xylosus, which were separated and identified from naturally fermented dried beef-Bundnerfleisch. The results were following:The result of the fermenting characteristics of the strains indicated that three strains have the properties to endure high salt and high nitrite;Lactobacillu plantarum and Pediococcus pentosaceus could affect the decomposition of protein, Staphylococcus xylosus could affect the decomposition of protein and fat;Lactobacillus plantarum and Pediococcus pentosaceus could inhibit the growths of some pathogenic bacteria, but Staphylococcus xylosus couldn't.The optimum mediums of these three bacteria were obtained. The optimum medium of Lactobacillus plantarum was based on the malt juice medium adding with 0.5% lactose, 1% peotone beef, 5% yeast extract,5% tomato juice;the optimum medium of Pediococcus pentosaceus was based on the malt juice medium adding with 0.5% lactose, 1.5% peotone beef, 5% yeast extract, 10% tomato juice;the optimum medium of Staphylococcus xylosus was based on the malt juice medium adding with 0.5% glocuse, 1.5% peotone beef, 0.1 %beef extract, 1 %sodium chloride.This paper studied the culture conditions of Lactobacillus plantarum, Pediococcus pentosaceus and Staphylococcus xylosus.The culture conditions of Lactobacillus plantarum and Pediococcrs pentosaceus were: cultured temperature 20℃, initial pH value 6.5~7.0, cultured time 20h;the culture condition of Staphylococcus xylosus was: cultured temperature 20℃, initial pH value 7.0, cultured time 20h. After incubation, the viable counts of three strains were 3.75×10~9cfu/mL, 3.68×10~9cfu/mLand 5.00×10~9cfu/mL.The concentrating enrichment conditions were studied and chosen, the results illstruted that the centrifugal effect was better than the bacteria filter;the effect of high rpm was helpful for bacteria's concentrating, improved the centrifugation concentrating times.The results indicated that the best effect could be obtained at 0~4℃ with 6 000rpm for 30min.Using 12% skimmed milk as suspended substrat, the function of protectors including
sucrose, glycerin was studied. Results suggested that sucrose, Vc, glycerin and CaCO3 had better effects on promoting cell count. According to orthogonal experiment, the optimal combined cryoprotectants of Lactobacillus plantarum, Pediococcus pentosaceus and Staphylococcus xylosus contained the following ingredients were based on the modified skimmed milk adding with 5% lactose, l%Vc, 5%glycerin and 3%CaCC>3;based on the modified skimmed milk adding with 4% lactose, l%Vc, 3%glycerin and 2%CaCC>3;based on the modified skimmed milk adding with 3%lactose, 1.5%Vc, 5%glycerin and 2%CaCO3. The survival of Lactobacillus plantarum, Pediococcus pentosaceus, Staphylococcus xylosus got to 63.01%, 66.20% and 70.47%;the cell suspension in freeze-drying culture starter containing the suitable protector might be lyophilized into the strains with high viability(the number of viable cell 2.62xl010cfu/g, 2.54x1010cfu/g and2.91xl010cfu/g).Measuring of quality guarantee-time: Storage effect under freezing condition and refrigeration condition was superior to those under indoor temperature, but the contrast was smallness.The prepared solid fermentation agents are stored under refrigeration condition for six months.
本文测定了植物乳杆菌、戊糖片球菌和木糖葡萄球菌的部分发酵特性,研究发现:三者均具有较强的耐盐性和耐亚硝酸盐性;植物乳杆菌和戊糖片球菌只具有蛋白降解能力,而木糖葡萄球菌具有蛋白降解能力和脂肪降解能力;植物乳杆菌和戊糖片球菌菌株对致病菌的抑菌效果较好,木糖葡萄球菌抑菌作用较差。
对植物乳杆菌、戊糖片球菌和木糖葡萄球菌的增殖培养基进行了优化,确定了菌株的复合增殖培养基配方。植物乳杆菌的复合增殖培养基配方为:麦芽汁培养基中添加2%乳糖、1%牛肉蛋白胨、2.5%酵母膏、10%番茄汁;戊糖片球菌的复合增殖培养基配方为:麦芽汁培养基中添加0.5%乳糖、1.5%牛肉蛋白胨、5%酵母膏、10%番茄汁;木糖葡萄球菌的复合增殖培养基配方为:麦芽汁培养基中添加0.5%葡萄糖、1.5%牛肉蛋白胨、0.1%牛肉膏、1%NaCl。
同时对菌株的增殖培养条件进行了研究,确定了菌株的培养条件。植物乳杆菌和戊糖片球菌:培养温度为20℃,培养基初始pH值6.5～7.0,培养时间20h,菌体密度可分别达3.75x10~9cfu/mL和3.68×10~9cfu/mL:木糖葡萄球菌:培养温度为20℃,培养基初始pH值7.0,培养时间20h,菌体密度可达5.00x10~9cfu/mL。
对菌体细胞浓缩分离条件进行了选择研究,结果显示:离心方法浓缩菌体的效果明显高于细菌过滤器的浓缩效果,并且采用高转速离心,可以提高浓缩菌体浓缩倍数,有利于菌体细胞富集分离。确定菌体细胞浓缩分离的适宜离心条件为0～4℃、6000rpm离心30min。
以12%的脱脂乳为冻干悬浮基质添加单因子制成冻干保护剂,进行冷冻干燥试验。从试验结果中发现蔗糖、维生素C、甘油和CaCO_3是植物乳杆菌、戊糖片球菌和木糖葡萄球菌的较好单因子保护剂。利用L_9(3~4)正交试验优化的植物乳杆菌、戊糖片球菌和木糖葡萄球菌的复合冻干保护剂配方分别为:在脱脂乳中添加5%蔗糖、0.5%维生素C、4%甘油、3%CACO_3;脱脂乳中添加4%蔗糖、1%维生素C、3%甘油、2%CACO_3和脱脂乳中添加3%蔗糖、1.5%维生素C、5%甘油、4%CACO_3,可分别使植物乳杆菌、戊糖片球菌和木糖葡萄球菌冷冻干燥存活率达到63.01%、66.20%和70.47%,制备的冻干菌粉发酵剂的活菌数分别为2.62×10~(10)cfu/g、2.54×10~(10)cfu/g和2.91×10~(10)cfu/g。
Biotechnological characteristics, high density cultivation, combined cryoprotectants of concentrated cultures production and quality guarantee-time of starts cultures.et al. were studied here systemically with three strains including Lactobacillus plantarum, Pediococcus pentosaceus and Staphylococcus xylosus, which were separated and identified from naturally fermented dried beef-Bundnerfleisch. The results were following:The result of the fermenting characteristics of the strains indicated that three strains have the properties to endure high salt and high nitrite;Lactobacillu plantarum and Pediococcus pentosaceus could affect the decomposition of protein, Staphylococcus xylosus could affect the decomposition of protein and fat;Lactobacillus plantarum and Pediococcus pentosaceus could inhibit the growths of some pathogenic bacteria, but Staphylococcus xylosus couldn't.The optimum mediums of these three bacteria were obtained. The optimum medium of Lactobacillus plantarum was based on the malt juice medium adding with 0.5% lactose, 1% peotone beef, 5% yeast extract,5% tomato juice;the optimum medium of Pediococcus pentosaceus was based on the malt juice medium adding with 0.5% lactose, 1.5% peotone beef, 5% yeast extract, 10% tomato juice;the optimum medium of Staphylococcus xylosus was based on the malt juice medium adding with 0.5% glocuse, 1.5% peotone beef, 0.1 %beef extract, 1 %sodium chloride.This paper studied the culture conditions of Lactobacillus plantarum, Pediococcus pentosaceus and Staphylococcus xylosus.The culture conditions of Lactobacillus plantarum and Pediococcrs pentosaceus were: cultured temperature 20℃, initial pH value 6.5~7.0, cultured time 20h;the culture condition of Staphylococcus xylosus was: cultured temperature 20℃, initial pH value 7.0, cultured time 20h. After incubation, the viable counts of three strains were 3.75×10~9cfu/mL, 3.68×10~9cfu/mLand 5.00×10~9cfu/mL.The concentrating enrichment conditions were studied and chosen, the results illstruted that the centrifugal effect was better than the bacteria filter;the effect of high rpm was helpful for bacteria's concentrating, improved the centrifugation concentrating times.The results indicated that the best effect could be obtained at 0~4℃ with 6 000rpm for 30min.Using 12% skimmed milk as suspended substrat, the function of protectors including
sucrose, glycerin was studied. Results suggested that sucrose, Vc, glycerin and CaCO3 had better effects on promoting cell count. According to orthogonal experiment, the optimal combined cryoprotectants of Lactobacillus plantarum, Pediococcus pentosaceus and Staphylococcus xylosus contained the following ingredients were based on the modified skimmed milk adding with 5% lactose, l%Vc, 5%glycerin and 3%CaCC>3;based on the modified skimmed milk adding with 4% lactose, l%Vc, 3%glycerin and 2%CaCC>3;based on the modified skimmed milk adding with 3%lactose, 1.5%Vc, 5%glycerin and 2%CaCO3. The survival of Lactobacillus plantarum, Pediococcus pentosaceus, Staphylococcus xylosus got to 63.01%, 66.20% and 70.47%;the cell suspension in freeze-drying culture starter containing the suitable protector might be lyophilized into the strains with high viability(the number of viable cell 2.62xl010cfu/g, 2.54x1010cfu/g and2.91xl010cfu/g).Measuring of quality guarantee-time: Storage effect under freezing condition and refrigeration condition was superior to those under indoor temperature, but the contrast was smallness.The prepared solid fermentation agents are stored under refrigeration condition for six months.
引文
[1] K Incze著.发酵干燥肉制品[J].肉类工业,1994(5):19～23.
[2] 马晓燕,南庆贤,等.干制牛肉火腿微生物研究进展[J].肉类研究,2001(1):34～35.
[3] Jessen B. Fermented meat products. St, Edmundsbury Press, 1995.
[4] Montel M C, Robinson R K. Fermented meat products. Encyclopedia of food microbiology, 2000, 3: 744-753. London: Academic Press.
[5] Cordero M R, Zumalacarregui J M. Characterization of Micrococcaceae isolated from salt used for Spanish dry-cured ham[J]. Letters in Applied Microbiology, 2000, 31: 303～306.
[6] 肖蓉,徐昆龙,刘旭川.云南牛干巴(腊牛肉) 中细菌的分离与鉴定[J].黄牛杂志,1998,24(3):21～22.
[7] Paleafi M A, Bersani C, Vittorio M M, et al. Effects of the curing and fermentation on the microflora of meat of various animal species[J]. Food Control, 2002, 13: 195-197.
[8] 周光宏,赵改名,等.我国传统腌腊肉制品存在的问题及对策[J].肉类研究,2003(1):3～7.
[9] 张美玲.如何利用发酵剂加工肉类制品[J].中国食品工业,2005,2:55～56.
[10] Reyes-Cano R, Dorantes-Alverez L, Hermandez-Sanchez H, et al. A traditional intermediated moisture meat: Beef Cecina[J]. Meat Science, 1994, 36: 365～370.
[11] Bacteriocinogenic activity from starter cultures used in Spanish meat industry[J]. Meat Science, 2002, 62: 237～243.
[12] Garcia I, Zumalacarregui J M, Diez V. Microbial succession and identification of micrococcaceae in dried beef cecina, a intermediated moisture meat product[J]. Food Microbiology, 1995, 12: 309～315
[13] Pinto M F. Charqui meats as fermented meat products: role of bacteria for some sensorial properties development[J]. Meat Science, 2002, 61: 187～191.
[14] 孙宝忠.干腌牛肉加工过程中微生物变化、作用及机理[D].博士论文,中国农业大学,2004.
[15] Elizabeth C, Gerald F F. Food fermentations: role of microorganisms in food production and preservation[J]. International Journal of Food Microbiology, 1999, 50: 131～149.
[16] Drosinos E H, Mataragas M, Xiraphi N, et al. Characterization of the microbial flora from a traditional Greek fermented sausage[J]. Meat Science, 2002, 69: 307～317.
[17] Sunesen L O, Stahnke L H. Mould starter cultures for dry sausages—selection, application and effects. Meat Science, 2003, 65: 935～948.
[18] Fausto G, Maria M, Maria A C. Use of Staphylococcus xylosus as a starter culture in dried sausages: effect on the biogenic amine content[J]. Meat Science, 2002, 61: 275～283.
[19] 李宗军,江汉湖.肉品微生物系统和肉类发酵剂研究[J].食品与发酵工业,2001,28(5):54～59.
[20] 王雪青,马长伟.发酵香肠及微生物发酵剂(一)[J].食品与发酵工业,1998,24(2):62～67.
[21] 宋照军,马汉军,潘润淑,等.发酵肉制品发酵剂的研究和应用[J].食品工业科技,1998,5: 68～70.
[22] 朱燕,罗欣.肉类发酵剂及其发酵方式[J].肉类研究,2003,1:13～15.
[23] 刘云鹤,何煜波.肉品发酵剂增殖培养基及培养条件的研究[J].湖南农业大学学报(自然科学版),2002,28(3):234～236.
[24] 刘云鹤.肉品干粉发酵剂的增殖培养及制备条件的研究[D].长沙,湖南农业大学,2002.
[25] Pidcock K, Heard G M, Henriksson A. Application of nontraditional meat starter cultures in production of Hungarian salami[J]: International Journal of Food Microbiology, 2002, 76: 75～81.
[26] Jim Bacus.微生物在肉类加工中的应用[M].张元生,许益民编译.北京:中国商业出版社,1991.
[27] 罗欣,朱燕.发酵剂微生物及其代谢与发酵香肠的工艺控制.食品与发酵工业,2001,28,3:67～71.
[28] Hammes W P, Knauf H. Starters in the processing of meat products[J]. Meat Science, 1994, 36: 155～168.
[29] Egon B H. Commercial bacterial starter cultures for fermented foods of the future[J]. Food Microbial, 2002, 78: 119～131.
[30] 王永霞.肉品发酵剂的菌种筛选及在发酵香肠中的应用[D].北京,中国农业大学,2004.
[31] 王海燕.发酵香肠的菌种分离、筛选及在发酵香肠中的应用研究[D].泰安,山东农业大学,2002.
[32] 吴祖兴,张华.发酵肉制品乳酸菌菌种筛选研究[J].食品科学,2002,23(9):47～49.
[33] 李凤彩,程文新,谢华,等.发酵香肠中菌种筛选标准探讨[J].食品科技,2002,6:78～79.
[34] Coppola R, Glaghacovo B. Charaterization of lactobacilli involved in the ripening of soppressata molisana, a typical southern Italy fermented sausage[J]. Food Microbiology, 1998, 15: 247～353.
[35] Hughes M C, Kerry J P, Arend E K. Characterization of proteolysis during the ripening of semi-dry fermented sausages[J]. Meat Science, 2002, 62: 205～216.
[36] Papamanoli E, Kotzekidou P, Tzanetakis N, et al. Characterization of Micrococcaceae isolated from dry fermented sausage[J]. Food Microbiology, 2002, 19: 441～449.
[37] 吕兵,张国农.分离自传统腊肠中的乳酸菌的特性研究[J].食品与发酵工业,2004,30(8):65～68.
[38] 何煜波.香肠发酵乳杆菌的基本特性[J].肉类工业,2002,6:22～26.
[39] 徐登峰,曹国文,戴荣国,等.植物乳酸菌对致病性大肠杆菌的抑制试验[J].四川畜牧兽医,2005,8:36～37.
[40] Geisen R, Locke F K, Krockel I O. Starter and protective cultures for meat and meat products[J]. Fleischwirtschaf, 1992, 72: 894～898.
[41] Papamanoli E, Tzanetakis E, Litopoulou T, et al. Characterization of lactic acid bacteria isolated from a Greek dry-fermented sausage in respect of their technological and probiotic properties[J]. Meat Science, 2003, 65: 859～867.
[42] Gardini F, Tofalo R, Suzzi G. A survey of antibiotic resistance in Micrococcaceae isolated from Italian dry fermented sausages[J]. Journal of Food Protection, 2003, 66: 937～945.
[43] 山丽杰,田洪涛,贾英民等.浓缩型乳酸菌发酵剂制备中几个技术关键问题的探讨[J],中国乳品工业,2002,30(5):66～69.
[44] 吕加平,梁占东,骆承庠,等.乳酸菌增菌培养基的优化设计[J].中国乳品工业,1999,(3):12～15.
[45] 谭欢,周传云.制备浓缩型乳酸菌发酵剂的研究进展.现代食品科技,2005,21(4):112～114.
[46] 吕兵,张国农.乳酸菌发酵剂浓缩培养的研究[J].中国乳品工业,2001,6:7～9.
[47] Gonzalez-Fandos M E, Sierra M, Garcoa-Lopez M L. The influence of manufacturing and drying conditions on the survival and toxinogenesis of Staphylococcus aureus in two Spanish dry sausages (chorizo and salchichon) [J]. Meat Science, 1999, 52: 411～419.
[48] Holzapfel W H. Appropriate starter culture technologies for small-scale fermentation in developing countries[J]. International Journal of Food Microbiology, 2002, 75: 197～212.
[49] 杨洁彬,凌代文,郭兴华,等.乳酸菌——生物学基础及应用[M].北京;中国轻工业出版社,1996,75～83:142～145.
[50] 王永霞,牛天贵.肉品发酵剂葡萄球菌和微球菌的分离筛选及其与乳酸菌混合培养的研究[J].肉类工业 2004,4:41～44.
[51] 李艾黎,代敏,霍贵成.浓缩型乳酸菌发酵剂的工业化生产[J].中国乳品工业,2005,33(6):50～53.
[52] 熊晓辉,于修,熊强,等.乳酸菌发酵剂高密度培养的研究[J].中国调味品,2004,(5):17～22.
[53] 刘丹,潘道东.直投式乳酸菌发酵剂增菌培养基的优化[J].食品科学,2005,26(9):204～207.
[54] 刘振民.乳酸菌高密度培养及浓缩型发酵剂研究[D].哈尔滨,东北农业大学,2002.
[55] 韦利军.肉类食品乳酸菌固体发酵剂的研制[D].沈阳,沈阳农业大学,2002.
[56] 奚霞,刘然,万守朋.直投式乳酸菌发酵剂的制备[J].食品研究与开发.2003,24(6):25～26.
[57] 田发益,钟国辉,纪素玲.乳酸菌在西藏的扩大培养和发酵试验.乳业科学与技术,2005(3):106～108.
[58] 黄君红,成洁珊,陈青荷.乳酸菌生长最佳培养基的筛选.中国酿造,2001,2:9～11.
[58] 乔东发,南庆贤,兰凤英.浓缩乳酸菌发酵剂制造工艺的研究[J].中国乳品工业,1998,26(24):13～15.
[59] Riesen D, Guthke R. High cell density cultivation of microorganisms[J]. Micrbiol Biotechol. 1999, 51: 422～430.
[60] 田洪涛,贾英民.嗜热链球菌真空冷冻干燥前后发酵活力变化[J].中国乳品工业,2001,29(6):17～19.
[61] 乔发东,吴秀芳.高效浓缩乳酸菌发酵剂的制作与应用.农牧产品开发,1998,1:32-35.
[62] 张建友,赵培城,徐静波,等.冷冻干燥酸奶发酵剂中菌体细胞的收集.粮油食品科技,2005,13(3):27-29.
[63] 张建友,赵培城,徐静波,等.真空冷冻干燥酸奶发酵剂的研究现状[J].河南工业大学学报,2005,26(2):86-89.
[64] 山丽杰.利用真空冷冻干燥技术制备高效浓缩型酸奶发酵剂的研究[D].保定,河北农业大学, 2004.
[65] 刘振民,骆承庠.乳酸菌冷冻损伤研究[J].食品与发酵工业,2002,(10):18～21.
[66] 黄良昌,吕晓玲,邢晓慧。酸奶发酵剂的研究进展[J].广州食品工业科技,2001,3:43～46.
[67] Tsvetan T. Studies the effects of low temperatures on lactic acid bacteria[J]. Cryobiology, 1982, 19: 211～214.
[68] 畅天狮,刘俊果,张桂,等.乳酸菌在酸性环境中的生理变化及pHin的调控机制.中国乳品工业,2002,30(2):7～10.
[69] 刘会平,南庆贤,乔发东.化学中和法制各高效浓缩乳酸菌发酵剂的研究[J].中国畜产与食品,1998,5(2):50～52.
[70] Brashears M M, Gilliland S E. Survival during frozen and subsequent refrigerated storage of lactobacillus acidophilus cells as influenced by the growth phase[J]. Journal of Dairy Science, 1995, 78: 2326～2335.
[71] 张建友,赵培城,徐静波,等.直投式酸奶发酵剂中高浓度细胞悬浮液的制备研究[J].现代食品科技,2005,21(1):9～13.
[72] 刘振民.采用微滤膜浓缩乳酸菌发酵液的工艺条件研究[J].食品发酵工业,2004,30(10):73～76.
[73] 卢行安,陈明生,袁杰利,等.乳酸菌冷冻干燥工艺的研究进展[J].中国微生态学杂志,2001,(1):58～61.
[74] Erandaf F. Method of quantifying the loss of acidification activity of lactic acid startersd uring freezing and frozens torage[J]. Dairy Research, 2000, 67: 83～90.
[75] 罗建玲,甘纯矶.保护荆对冷冻干燥过程中乳酸菌特性影响的研究[J].福建轻纺,2005,5:7～11.
[76] 陈声明,吕琴.微生物冷冻干燥的抗性机理[J].微生物学通报,1996,(4):236～238.
[77] 乔东发.冷冻干燥浓缩乳酸菌发酵剂抗冷冻保护剂的选择[J].河南职技师院学报,1998,3:43～46.
[78] 蒲丽丽,刘宁,张英华,等.乳酸菌冻干保护剂及保护机理的研究进展[J].现代食品科技,2004,24(1):147～150.
[79] 邵伟,熊泽,乐超银.冻干法制备乳酸菌发酵剂工艺条件研究[J].三峡大学学报,2005,27(3):173～273.
[80] 袁亚宏,岳田利,高振鹏,等.冻干高活力乳酸菌粉保护剂的研究[J].西北农林科技大学学报,2003,31:82～85.
[81] 戴秀玉,沈义国,周坚.海藻糖对乳酸菌的抗逆保护作用[J].微生物学通报,2001,28(2):46～50.
[82] 蒲丽丽,刘宁,张英华,等.乳酸菌冻干损伤与保护的研究进展[J].食品工业科技,2005,26(7):182～184.
[83] 李春,张兰威.乳酸菌细胞膜冻干损伤研究进展[J].中国乳品工业,2004,32(3):28～30.
[84] 吕加平,骆承庠.冻干保护剂的研究及应用[A].第三届海峡两岸畜产品加工学术研讨会论文集[C].1996,42～50.
[85] 张建国,张春晖,李凤彩,等.乳酸菌冻干伤害与保护[J].食品工业科技,2003,24(1):10～12.
[86] 王宇.冷冻干燥技术生产直投式酸奶发酵剂的研究[D].哈尔滨,东北农业大学,2000.
[87] 黄良昌.真空冷冻干燥法生产干酸奶发酵剂工艺的研究[D].天津,天津轻工业学院,2002.
[88] 谢丽源.活性干燥乳酸菌发酵剂的研究[D].重庆,西南农业大学,2003.
[89] 张兰威,鄂志强,万海峰等.乳酸菌增菌培养基的筛选及干燥保护剂的选择[J].中国乳品工业,2000,(2):7～9.
[90] Thunell R K. Frozen starters from internal-pH-control-grow-culture[J]. Jounal of dairy Science, 1984, 67: 24～36.
[91] Madden U A. Effects of bacterium coprostanoligenes and lactobacillus on pH lipcontent, and cholesterol of fermented pork and mutton sausage[J]. Institute of Food Tenhnologists, 1999, 64(5): 544～547.
[92] 刘玺.乳酸菌发酵中式香肠发色效果研究[J].肉类工业,2000(9):16～19.
[93] 乔东发.超滤技术制备浓缩乳酸菌发酵剂的研究[J].中国畜产与食品,1998,5(3):112～113.
[94] 吕兵,张国农,林金资.酸奶发酵剂速效干燥制备中保护剂的研究[J].中国乳品工业,1996,2:3～5.
[95] Bumsoo H, John C. Direct cell injury associated with eutectic crystllization during freezing[J]. Cryobiology, 2004, 48: 8～21.
[96] Antoni G. Trehalose, a cryoproectant for lactobacillus bulgaricus[J]. Cryobiology, 1989, 26: 149～153
[97] Trsic N, Kodzic A, Baras J, et al. The influence of a cryoprotective medium containing glycerol on the lyophilization of lactic acid bacteria[J]. Chem. Soc, 2001, 66(7): 435～441.
[98] 史媛英,肖冬光.酸奶发酵剂高浓度培养的研究[J].天津轻工业学院学报,1999,1:20～25.
[99] Graciela F D V. Comparative study of the efficiency of sone additives in protecting lactic acid bacteria against freeze-drying[J]. Cryobilolgy, 1983, 20: 560～566.
[100] Costa E. Effect of protective agents, rehydration media and initial cell concentration on viability of pantoea agglom-erans strain CPA-2 subiected to freeze-drying[J]. Journal of Applied Microbiology. 2000, 89: 793～800.
[101] 吕为群,骆承庠,刘书臣.乳酸菌在冷冻干燥过程中存活率影响因素探讨[J].中国乳品工业,1993,(5):217～220.
[102] Zdenek H. Protectants used in the crypreservation of microorganisms[J]. Cryobiology 2003, 46: 205～229.
[103] 黄君红,成洁珊,陈青荷.乳酸菌生产最佳培养基的筛选[J].中国酿造,2001,2:9～11.
[104] 刘慧,李铁晶主编.新编食品微生物学实验指导[M].东北农业大学,2000.
[105] Brian J.B.W.徐岩译.发酵食品微生物学(第二版).北京:中国轻工业出版社,2001.
[106] Paul B C, Danforth P M, Peter R C, et al. Stabilizationa ndp reservation of L actobacilusa cidophilusin saccharide matrices[J]. Cryobiology, 2000, 41: 17～24.
[107] Champagne C P, Mondou F. Effect of polymers and storage temperature on the stability of freeze-dried lactic acid bacteria[J]. Food-resint, 1996(29): 555-562
[108]Rahmab B. Effect of calciumon the cryopreservation of lactobacillus bulgaricus in different freezing media[J]. Cryobiology, 1990, 27: 336-342 .
[109]Tan C S, Vaningen C W, Talsma H, et al. Freeze-drying of composition and glass transition temperature of the protect[J]. Cryobiology, 1995, 32: 60-67.
[110]Champagne C P, Gardner N J. The effect of protective ingredients on the survidal of immobilized cells of Streptococcus thermophilus to air and freeze-drying[J]. Electronic Biotechnology. 2001. 3(4): 1-7.
[111] Beal C, Fonseca F, Corrieu G. Resistance to freezing and frozen storage of streptococcus thermophilus is related to membrane fatty acid composition[J]. Jounal Dairy Science, 2001, 84: 2347-2356.
[112]Taa G S. Freeze-drying of composition and glass transition temperature of the protectant[J] Cryobiology, 1995, 32: 60-67.
[113]Graaciela F D V. Comparative study of the efficiency of some additives in protecting lactic acid bacteria against freeze-drying[J]. Cryobiology, 1983, 20: 560-566.
[114]Sinha R P. Effect of catbohydrate on the vibility of streptococcus thermophilus[J]. 1978, 7: 537-541.
[115]Biondl A C. Permeability of lipid membranes revised in relation to freeze-thaw processes[J] 1992, 29: 323-331.
[116]Osborne R J W. Production of frozen concentrated cheese starters by diffusion culture[J]. Journal of Food Science, 1991, 57(5): 1779-1784.
[117]Smittle R B. Death of lactobacillus bulgaricus resulting from liquid nitrogen freezing[J]. Microbiol, 1972, 24: 55.
[118]Kilara A. Effect of cryopotective agents on freeze drying and storage of lactic culture[J]. Dairy, 1976, 11: 8.
[2] 马晓燕,南庆贤,等.干制牛肉火腿微生物研究进展[J].肉类研究,2001(1):34～35.
[3] Jessen B. Fermented meat products. St, Edmundsbury Press, 1995.
[4] Montel M C, Robinson R K. Fermented meat products. Encyclopedia of food microbiology, 2000, 3: 744-753. London: Academic Press.
[5] Cordero M R, Zumalacarregui J M. Characterization of Micrococcaceae isolated from salt used for Spanish dry-cured ham[J]. Letters in Applied Microbiology, 2000, 31: 303～306.
[6] 肖蓉,徐昆龙,刘旭川.云南牛干巴(腊牛肉) 中细菌的分离与鉴定[J].黄牛杂志,1998,24(3):21～22.
[7] Paleafi M A, Bersani C, Vittorio M M, et al. Effects of the curing and fermentation on the microflora of meat of various animal species[J]. Food Control, 2002, 13: 195-197.
[8] 周光宏,赵改名,等.我国传统腌腊肉制品存在的问题及对策[J].肉类研究,2003(1):3～7.
[9] 张美玲.如何利用发酵剂加工肉类制品[J].中国食品工业,2005,2:55～56.
[10] Reyes-Cano R, Dorantes-Alverez L, Hermandez-Sanchez H, et al. A traditional intermediated moisture meat: Beef Cecina[J]. Meat Science, 1994, 36: 365～370.
[11] Bacteriocinogenic activity from starter cultures used in Spanish meat industry[J]. Meat Science, 2002, 62: 237～243.
[12] Garcia I, Zumalacarregui J M, Diez V. Microbial succession and identification of micrococcaceae in dried beef cecina, a intermediated moisture meat product[J]. Food Microbiology, 1995, 12: 309～315
[13] Pinto M F. Charqui meats as fermented meat products: role of bacteria for some sensorial properties development[J]. Meat Science, 2002, 61: 187～191.
[14] 孙宝忠.干腌牛肉加工过程中微生物变化、作用及机理[D].博士论文,中国农业大学,2004.
[15] Elizabeth C, Gerald F F. Food fermentations: role of microorganisms in food production and preservation[J]. International Journal of Food Microbiology, 1999, 50: 131～149.
[16] Drosinos E H, Mataragas M, Xiraphi N, et al. Characterization of the microbial flora from a traditional Greek fermented sausage[J]. Meat Science, 2002, 69: 307～317.
[17] Sunesen L O, Stahnke L H. Mould starter cultures for dry sausages—selection, application and effects. Meat Science, 2003, 65: 935～948.
[18] Fausto G, Maria M, Maria A C. Use of Staphylococcus xylosus as a starter culture in dried sausages: effect on the biogenic amine content[J]. Meat Science, 2002, 61: 275～283.
[19] 李宗军,江汉湖.肉品微生物系统和肉类发酵剂研究[J].食品与发酵工业,2001,28(5):54～59.
[20] 王雪青,马长伟.发酵香肠及微生物发酵剂(一)[J].食品与发酵工业,1998,24(2):62～67.
[21] 宋照军,马汉军,潘润淑,等.发酵肉制品发酵剂的研究和应用[J].食品工业科技,1998,5: 68～70.
[22] 朱燕,罗欣.肉类发酵剂及其发酵方式[J].肉类研究,2003,1:13～15.
[23] 刘云鹤,何煜波.肉品发酵剂增殖培养基及培养条件的研究[J].湖南农业大学学报(自然科学版),2002,28(3):234～236.
[24] 刘云鹤.肉品干粉发酵剂的增殖培养及制备条件的研究[D].长沙,湖南农业大学,2002.
[25] Pidcock K, Heard G M, Henriksson A. Application of nontraditional meat starter cultures in production of Hungarian salami[J]: International Journal of Food Microbiology, 2002, 76: 75～81.
[26] Jim Bacus.微生物在肉类加工中的应用[M].张元生,许益民编译.北京:中国商业出版社,1991.
[27] 罗欣,朱燕.发酵剂微生物及其代谢与发酵香肠的工艺控制.食品与发酵工业,2001,28,3:67～71.
[28] Hammes W P, Knauf H. Starters in the processing of meat products[J]. Meat Science, 1994, 36: 155～168.
[29] Egon B H. Commercial bacterial starter cultures for fermented foods of the future[J]. Food Microbial, 2002, 78: 119～131.
[30] 王永霞.肉品发酵剂的菌种筛选及在发酵香肠中的应用[D].北京,中国农业大学,2004.
[31] 王海燕.发酵香肠的菌种分离、筛选及在发酵香肠中的应用研究[D].泰安,山东农业大学,2002.
[32] 吴祖兴,张华.发酵肉制品乳酸菌菌种筛选研究[J].食品科学,2002,23(9):47～49.
[33] 李凤彩,程文新,谢华,等.发酵香肠中菌种筛选标准探讨[J].食品科技,2002,6:78～79.
[34] Coppola R, Glaghacovo B. Charaterization of lactobacilli involved in the ripening of soppressata molisana, a typical southern Italy fermented sausage[J]. Food Microbiology, 1998, 15: 247～353.
[35] Hughes M C, Kerry J P, Arend E K. Characterization of proteolysis during the ripening of semi-dry fermented sausages[J]. Meat Science, 2002, 62: 205～216.
[36] Papamanoli E, Kotzekidou P, Tzanetakis N, et al. Characterization of Micrococcaceae isolated from dry fermented sausage[J]. Food Microbiology, 2002, 19: 441～449.
[37] 吕兵,张国农.分离自传统腊肠中的乳酸菌的特性研究[J].食品与发酵工业,2004,30(8):65～68.
[38] 何煜波.香肠发酵乳杆菌的基本特性[J].肉类工业,2002,6:22～26.
[39] 徐登峰,曹国文,戴荣国,等.植物乳酸菌对致病性大肠杆菌的抑制试验[J].四川畜牧兽医,2005,8:36～37.
[40] Geisen R, Locke F K, Krockel I O. Starter and protective cultures for meat and meat products[J]. Fleischwirtschaf, 1992, 72: 894～898.
[41] Papamanoli E, Tzanetakis E, Litopoulou T, et al. Characterization of lactic acid bacteria isolated from a Greek dry-fermented sausage in respect of their technological and probiotic properties[J]. Meat Science, 2003, 65: 859～867.
[42] Gardini F, Tofalo R, Suzzi G. A survey of antibiotic resistance in Micrococcaceae isolated from Italian dry fermented sausages[J]. Journal of Food Protection, 2003, 66: 937～945.
[43] 山丽杰,田洪涛,贾英民等.浓缩型乳酸菌发酵剂制备中几个技术关键问题的探讨[J],中国乳品工业,2002,30(5):66～69.
[44] 吕加平,梁占东,骆承庠,等.乳酸菌增菌培养基的优化设计[J].中国乳品工业,1999,(3):12～15.
[45] 谭欢,周传云.制备浓缩型乳酸菌发酵剂的研究进展.现代食品科技,2005,21(4):112～114.
[46] 吕兵,张国农.乳酸菌发酵剂浓缩培养的研究[J].中国乳品工业,2001,6:7～9.
[47] Gonzalez-Fandos M E, Sierra M, Garcoa-Lopez M L. The influence of manufacturing and drying conditions on the survival and toxinogenesis of Staphylococcus aureus in two Spanish dry sausages (chorizo and salchichon) [J]. Meat Science, 1999, 52: 411～419.
[48] Holzapfel W H. Appropriate starter culture technologies for small-scale fermentation in developing countries[J]. International Journal of Food Microbiology, 2002, 75: 197～212.
[49] 杨洁彬,凌代文,郭兴华,等.乳酸菌——生物学基础及应用[M].北京;中国轻工业出版社,1996,75～83:142～145.
[50] 王永霞,牛天贵.肉品发酵剂葡萄球菌和微球菌的分离筛选及其与乳酸菌混合培养的研究[J].肉类工业 2004,4:41～44.
[51] 李艾黎,代敏,霍贵成.浓缩型乳酸菌发酵剂的工业化生产[J].中国乳品工业,2005,33(6):50～53.
[52] 熊晓辉,于修,熊强,等.乳酸菌发酵剂高密度培养的研究[J].中国调味品,2004,(5):17～22.
[53] 刘丹,潘道东.直投式乳酸菌发酵剂增菌培养基的优化[J].食品科学,2005,26(9):204～207.
[54] 刘振民.乳酸菌高密度培养及浓缩型发酵剂研究[D].哈尔滨,东北农业大学,2002.
[55] 韦利军.肉类食品乳酸菌固体发酵剂的研制[D].沈阳,沈阳农业大学,2002.
[56] 奚霞,刘然,万守朋.直投式乳酸菌发酵剂的制备[J].食品研究与开发.2003,24(6):25～26.
[57] 田发益,钟国辉,纪素玲.乳酸菌在西藏的扩大培养和发酵试验.乳业科学与技术,2005(3):106～108.
[58] 黄君红,成洁珊,陈青荷.乳酸菌生长最佳培养基的筛选.中国酿造,2001,2:9～11.
[58] 乔东发,南庆贤,兰凤英.浓缩乳酸菌发酵剂制造工艺的研究[J].中国乳品工业,1998,26(24):13～15.
[59] Riesen D, Guthke R. High cell density cultivation of microorganisms[J]. Micrbiol Biotechol. 1999, 51: 422～430.
[60] 田洪涛,贾英民.嗜热链球菌真空冷冻干燥前后发酵活力变化[J].中国乳品工业,2001,29(6):17～19.
[61] 乔发东,吴秀芳.高效浓缩乳酸菌发酵剂的制作与应用.农牧产品开发,1998,1:32-35.
[62] 张建友,赵培城,徐静波,等.冷冻干燥酸奶发酵剂中菌体细胞的收集.粮油食品科技,2005,13(3):27-29.
[63] 张建友,赵培城,徐静波,等.真空冷冻干燥酸奶发酵剂的研究现状[J].河南工业大学学报,2005,26(2):86-89.
[64] 山丽杰.利用真空冷冻干燥技术制备高效浓缩型酸奶发酵剂的研究[D].保定,河北农业大学, 2004.
[65] 刘振民,骆承庠.乳酸菌冷冻损伤研究[J].食品与发酵工业,2002,(10):18～21.
[66] 黄良昌,吕晓玲,邢晓慧。酸奶发酵剂的研究进展[J].广州食品工业科技,2001,3:43～46.
[67] Tsvetan T. Studies the effects of low temperatures on lactic acid bacteria[J]. Cryobiology, 1982, 19: 211～214.
[68] 畅天狮,刘俊果,张桂,等.乳酸菌在酸性环境中的生理变化及pHin的调控机制.中国乳品工业,2002,30(2):7～10.
[69] 刘会平,南庆贤,乔发东.化学中和法制各高效浓缩乳酸菌发酵剂的研究[J].中国畜产与食品,1998,5(2):50～52.
[70] Brashears M M, Gilliland S E. Survival during frozen and subsequent refrigerated storage of lactobacillus acidophilus cells as influenced by the growth phase[J]. Journal of Dairy Science, 1995, 78: 2326～2335.
[71] 张建友,赵培城,徐静波,等.直投式酸奶发酵剂中高浓度细胞悬浮液的制备研究[J].现代食品科技,2005,21(1):9～13.
[72] 刘振民.采用微滤膜浓缩乳酸菌发酵液的工艺条件研究[J].食品发酵工业,2004,30(10):73～76.
[73] 卢行安,陈明生,袁杰利,等.乳酸菌冷冻干燥工艺的研究进展[J].中国微生态学杂志,2001,(1):58～61.
[74] Erandaf F. Method of quantifying the loss of acidification activity of lactic acid startersd uring freezing and frozens torage[J]. Dairy Research, 2000, 67: 83～90.
[75] 罗建玲,甘纯矶.保护荆对冷冻干燥过程中乳酸菌特性影响的研究[J].福建轻纺,2005,5:7～11.
[76] 陈声明,吕琴.微生物冷冻干燥的抗性机理[J].微生物学通报,1996,(4):236～238.
[77] 乔东发.冷冻干燥浓缩乳酸菌发酵剂抗冷冻保护剂的选择[J].河南职技师院学报,1998,3:43～46.
[78] 蒲丽丽,刘宁,张英华,等.乳酸菌冻干保护剂及保护机理的研究进展[J].现代食品科技,2004,24(1):147～150.
[79] 邵伟,熊泽,乐超银.冻干法制备乳酸菌发酵剂工艺条件研究[J].三峡大学学报,2005,27(3):173～273.
[80] 袁亚宏,岳田利,高振鹏,等.冻干高活力乳酸菌粉保护剂的研究[J].西北农林科技大学学报,2003,31:82～85.
[81] 戴秀玉,沈义国,周坚.海藻糖对乳酸菌的抗逆保护作用[J].微生物学通报,2001,28(2):46～50.
[82] 蒲丽丽,刘宁,张英华,等.乳酸菌冻干损伤与保护的研究进展[J].食品工业科技,2005,26(7):182～184.
[83] 李春,张兰威.乳酸菌细胞膜冻干损伤研究进展[J].中国乳品工业,2004,32(3):28～30.
[84] 吕加平,骆承庠.冻干保护剂的研究及应用[A].第三届海峡两岸畜产品加工学术研讨会论文集[C].1996,42～50.
[85] 张建国,张春晖,李凤彩,等.乳酸菌冻干伤害与保护[J].食品工业科技,2003,24(1):10～12.
[86] 王宇.冷冻干燥技术生产直投式酸奶发酵剂的研究[D].哈尔滨,东北农业大学,2000.
[87] 黄良昌.真空冷冻干燥法生产干酸奶发酵剂工艺的研究[D].天津,天津轻工业学院,2002.
[88] 谢丽源.活性干燥乳酸菌发酵剂的研究[D].重庆,西南农业大学,2003.
[89] 张兰威,鄂志强,万海峰等.乳酸菌增菌培养基的筛选及干燥保护剂的选择[J].中国乳品工业,2000,(2):7～9.
[90] Thunell R K. Frozen starters from internal-pH-control-grow-culture[J]. Jounal of dairy Science, 1984, 67: 24～36.
[91] Madden U A. Effects of bacterium coprostanoligenes and lactobacillus on pH lipcontent, and cholesterol of fermented pork and mutton sausage[J]. Institute of Food Tenhnologists, 1999, 64(5): 544～547.
[92] 刘玺.乳酸菌发酵中式香肠发色效果研究[J].肉类工业,2000(9):16～19.
[93] 乔东发.超滤技术制备浓缩乳酸菌发酵剂的研究[J].中国畜产与食品,1998,5(3):112～113.
[94] 吕兵,张国农,林金资.酸奶发酵剂速效干燥制备中保护剂的研究[J].中国乳品工业,1996,2:3～5.
[95] Bumsoo H, John C. Direct cell injury associated with eutectic crystllization during freezing[J]. Cryobiology, 2004, 48: 8～21.
[96] Antoni G. Trehalose, a cryoproectant for lactobacillus bulgaricus[J]. Cryobiology, 1989, 26: 149～153
[97] Trsic N, Kodzic A, Baras J, et al. The influence of a cryoprotective medium containing glycerol on the lyophilization of lactic acid bacteria[J]. Chem. Soc, 2001, 66(7): 435～441.
[98] 史媛英,肖冬光.酸奶发酵剂高浓度培养的研究[J].天津轻工业学院学报,1999,1:20～25.
[99] Graciela F D V. Comparative study of the efficiency of sone additives in protecting lactic acid bacteria against freeze-drying[J]. Cryobilolgy, 1983, 20: 560～566.
[100] Costa E. Effect of protective agents, rehydration media and initial cell concentration on viability of pantoea agglom-erans strain CPA-2 subiected to freeze-drying[J]. Journal of Applied Microbiology. 2000, 89: 793～800.
[101] 吕为群,骆承庠,刘书臣.乳酸菌在冷冻干燥过程中存活率影响因素探讨[J].中国乳品工业,1993,(5):217～220.
[102] Zdenek H. Protectants used in the crypreservation of microorganisms[J]. Cryobiology 2003, 46: 205～229.
[103] 黄君红,成洁珊,陈青荷.乳酸菌生产最佳培养基的筛选[J].中国酿造,2001,2:9～11.
[104] 刘慧,李铁晶主编.新编食品微生物学实验指导[M].东北农业大学,2000.
[105] Brian J.B.W.徐岩译.发酵食品微生物学(第二版).北京:中国轻工业出版社,2001.
[106] Paul B C, Danforth P M, Peter R C, et al. Stabilizationa ndp reservation of L actobacilusa cidophilusin saccharide matrices[J]. Cryobiology, 2000, 41: 17～24.
[107] Champagne C P, Mondou F. Effect of polymers and storage temperature on the stability of freeze-dried lactic acid bacteria[J]. Food-resint, 1996(29): 555-562
[108]Rahmab B. Effect of calciumon the cryopreservation of lactobacillus bulgaricus in different freezing media[J]. Cryobiology, 1990, 27: 336-342 .
[109]Tan C S, Vaningen C W, Talsma H, et al. Freeze-drying of composition and glass transition temperature of the protect[J]. Cryobiology, 1995, 32: 60-67.
[110]Champagne C P, Gardner N J. The effect of protective ingredients on the survidal of immobilized cells of Streptococcus thermophilus to air and freeze-drying[J]. Electronic Biotechnology. 2001. 3(4): 1-7.
[111] Beal C, Fonseca F, Corrieu G. Resistance to freezing and frozen storage of streptococcus thermophilus is related to membrane fatty acid composition[J]. Jounal Dairy Science, 2001, 84: 2347-2356.
[112]Taa G S. Freeze-drying of composition and glass transition temperature of the protectant[J] Cryobiology, 1995, 32: 60-67.
[113]Graaciela F D V. Comparative study of the efficiency of some additives in protecting lactic acid bacteria against freeze-drying[J]. Cryobiology, 1983, 20: 560-566.
[114]Sinha R P. Effect of catbohydrate on the vibility of streptococcus thermophilus[J]. 1978, 7: 537-541.
[115]Biondl A C. Permeability of lipid membranes revised in relation to freeze-thaw processes[J] 1992, 29: 323-331.
[116]Osborne R J W. Production of frozen concentrated cheese starters by diffusion culture[J]. Journal of Food Science, 1991, 57(5): 1779-1784.
[117]Smittle R B. Death of lactobacillus bulgaricus resulting from liquid nitrogen freezing[J]. Microbiol, 1972, 24: 55.
[118]Kilara A. Effect of cryopotective agents on freeze drying and storage of lactic culture[J]. Dairy, 1976, 11: 8.