嗜酸乳杆菌和干酪乳杆菌β-半乳糖苷酶和蛋白水解活性的研究
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摘要
本课题对来源于蒙古共和国和内蒙古锡林郭勒盟牧民家酸马奶酒样品中分离鉴定得到的6株嗜酸乳杆菌和5株干酪乳杆菌的?－半乳糖苷酶活性、蛋白水解活性及其在发酵乳中的部分生物学特性进行了初步研究，主要研究结果如下：
嗜酸乳杆菌和干酪乳杆菌在液体MRS培养基中37℃，18小时培养后均表现出不同程度的?－半乳糖苷酶活力，嗜酸乳杆菌的的乳糖酶活力单位为0.767~5.602u，平均酶活2.217u，干酪乳杆菌的乳糖酶活力单位为0.211~3.312u，平均酶活1.070u。菌种之间有差异，菌株之间也存在差异。
嗜酸乳杆菌和干酪乳杆菌具有一定的蛋白水解活性，pH、温度、反应时间都会对其蛋白水解活力产生影响。不同菌株蛋白水解作用的最适pH有所不同。在本实验温度范围内，温度升高有利于蛋白水解。
嗜酸乳杆菌和干酪乳杆菌发酵乳及其冷藏保存的研究表明，在37℃发酵过程中，嗜酸乳杆菌和干酪乳杆菌的产酸量平稳上升，发酵乳中游离氨基酸的量有不同程度的增加。24小时发酵结束后，嗜酸乳杆菌发酵乳平均酸度为115.5゜T，pH值变化在3.71~4.47之间，乳中酪氨酸含量为95.7ug/ml~135.15ug/ml；干酪乳杆菌发酵乳平均酸度为99.36゜T，pH值变化在3.86~4.83之间，乳中酪氨酸含量为94.8ug/ml~130.05ug/ml，供试菌株全部凝乳并有程度不同的芳香气味。在4℃冷藏保存的过程中，嗜酸乳杆菌和干酪乳杆菌保持了一部分蛋白水解活力，发酵乳中游离氨基酸的量继续增加。随着发酵乳pH的降低，供试菌株的?－半乳糖苷酶活性下降，宏观表现为产酸量继续增加，但变化趋于平缓。且在发酵乳保存到21天时，嗜酸乳杆菌和干酪乳杆菌具有较高的存活率，活菌数均高于108cfu/ml。
The subject of present research is ?-galactosidase, proteolytic activities and some biological characteristics of them in yoghurt of six strains of Lactobacillus acidophilus and five strains of Lactobacillus casei.Strains were isolated from the koumiss samples collected from the heardsman,s house in Mongolia and Inner Mongolia – Xilingole District .
The ?-galactosidase activities of Lactobacillus acidophilus and Lactobacillus casei were different. The activity range of ?-galactosidase was 0.767u~5.602u, and the average activity is 2.217u for the former. The range activity of ?-galactosidase was 0.211u~3.312u, and the average activity was 1.070u for the latter. Proteolytic activities varied among strains of L.acidophilus and L.casei .Temperature , pH , and reaction time affected the proteolytic activities. The maximal proteolytic activities of individual strains were different .With the increasing of the temperature the proteolytic activities of both strains were rising in this reseach. Some biological characteristics of the both cultures incubated in skimed milk at 37℃for 24 hours and then stored at 4 ℃for 21 days were studied . During the fermentation at 37℃ for 24 hours ,a significant increasing of acid production and soluble trysine in yoghurt were detected . At the end of fermentation , the average titration acidity of yoghurt inoculated L.acidophilus and L.casei were 115.5゜T and 99.36゜T respectively, correspondingly pH varied among 3.71~4.47 and 3.86~4.83 .The concentration range of soluble trysine was 95.70ug/ml ~ 135.15mg/ml for the former,and 94.80 ug/ml ~ 130.05 mg/ml for the latter. Some of ?-galactosidase and proteolytic activities of both strains were maintained during the observation stored at 4℃ for 21 days .L.acidophilus and L.casei both kept relatively high viability (>108cfu/ml) until the 21th day.
嗜酸乳杆菌和干酪乳杆菌在液体MRS培养基中37℃，18小时培养后均表现出不同程度的?－半乳糖苷酶活力，嗜酸乳杆菌的的乳糖酶活力单位为0.767~5.602u，平均酶活2.217u，干酪乳杆菌的乳糖酶活力单位为0.211~3.312u，平均酶活1.070u。菌种之间有差异，菌株之间也存在差异。
嗜酸乳杆菌和干酪乳杆菌具有一定的蛋白水解活性，pH、温度、反应时间都会对其蛋白水解活力产生影响。不同菌株蛋白水解作用的最适pH有所不同。在本实验温度范围内，温度升高有利于蛋白水解。
嗜酸乳杆菌和干酪乳杆菌发酵乳及其冷藏保存的研究表明，在37℃发酵过程中，嗜酸乳杆菌和干酪乳杆菌的产酸量平稳上升，发酵乳中游离氨基酸的量有不同程度的增加。24小时发酵结束后，嗜酸乳杆菌发酵乳平均酸度为115.5゜T，pH值变化在3.71~4.47之间，乳中酪氨酸含量为95.7ug/ml~135.15ug/ml；干酪乳杆菌发酵乳平均酸度为99.36゜T，pH值变化在3.86~4.83之间，乳中酪氨酸含量为94.8ug/ml~130.05ug/ml，供试菌株全部凝乳并有程度不同的芳香气味。在4℃冷藏保存的过程中，嗜酸乳杆菌和干酪乳杆菌保持了一部分蛋白水解活力，发酵乳中游离氨基酸的量继续增加。随着发酵乳pH的降低，供试菌株的?－半乳糖苷酶活性下降，宏观表现为产酸量继续增加，但变化趋于平缓。且在发酵乳保存到21天时，嗜酸乳杆菌和干酪乳杆菌具有较高的存活率，活菌数均高于108cfu/ml。
The subject of present research is ?-galactosidase, proteolytic activities and some biological characteristics of them in yoghurt of six strains of Lactobacillus acidophilus and five strains of Lactobacillus casei.Strains were isolated from the koumiss samples collected from the heardsman,s house in Mongolia and Inner Mongolia – Xilingole District .
The ?-galactosidase activities of Lactobacillus acidophilus and Lactobacillus casei were different. The activity range of ?-galactosidase was 0.767u~5.602u, and the average activity is 2.217u for the former. The range activity of ?-galactosidase was 0.211u~3.312u, and the average activity was 1.070u for the latter. Proteolytic activities varied among strains of L.acidophilus and L.casei .Temperature , pH , and reaction time affected the proteolytic activities. The maximal proteolytic activities of individual strains were different .With the increasing of the temperature the proteolytic activities of both strains were rising in this reseach. Some biological characteristics of the both cultures incubated in skimed milk at 37℃for 24 hours and then stored at 4 ℃for 21 days were studied . During the fermentation at 37℃ for 24 hours ,a significant increasing of acid production and soluble trysine in yoghurt were detected . At the end of fermentation , the average titration acidity of yoghurt inoculated L.acidophilus and L.casei were 115.5゜T and 99.36゜T respectively, correspondingly pH varied among 3.71~4.47 and 3.86~4.83 .The concentration range of soluble trysine was 95.70ug/ml ~ 135.15mg/ml for the former,and 94.80 ug/ml ~ 130.05 mg/ml for the latter. Some of ?-galactosidase and proteolytic activities of both strains were maintained during the observation stored at 4℃ for 21 days .L.acidophilus and L.casei both kept relatively high viability (>108cfu/ml) until the 21th day.
引文
1 顾瑞霞主编.乳酸菌与人体保健[M].北京：科学出版社.1995:1~155.
Fuller R . Probiotics: an Overview . Human Health : The Contribution of Microorgnisms [M]. 1992:63~73.
骆承庠主编.乳与乳制品工艺学[M] . 北京：农业出版社 , 1992: 39.
顾瑞霞主编.乳与乳制品的生理功能特性[M].北京: 中国轻工业出版社.2000: 294~460.
Bayless ,T . M. , N. S. Rosensweig .A racial diffrernce in incidience of lactose deficiency : A survrey of milk intolerance and lactase deficiency in healthy adult males [J]. American Jouenal Medicine Assay . 1966 , 197 : 968.
Haemmerli ,U .P. ,H .Kistler , R . Amman ,T.Marthaler , G . Semenza , S. Auricchio , A . Prader . Acquired milk intolerance in the adult caused by lactose intestinal lactase activity [J]. American Journal Medicine . 1965 , 38 : 7.
Littman, A ., J .B . Hammond . Diarrhea in adult cause by deficiency in intestinal disaccharidase [J] . Gastroenterology . 1965 . 48 : 237.
Schaafsma, G. ,p . , P . R . Dekker , H . Warrd . Microbial lactase activity and digestion of lactose [J]. Nethessland Milk Dairy . 1998 . 42 : 121.
Norton ,S ., M. D. Rosensweig . Adult human milk intolerance and intestinal lactase deficiency [J]. Dairy Science 1959 , 52 (5) : 585~587.
Dennis ,A, Savaiano . The influence and microbe-containing dairy foods [J]. Dairy Science . 1987,70:397~406.
B.A.费林得. 酸奶菌种选择对风味、抗微生物活性、β-半乳糖苷酶和B族维生素的影响. 中国乳品工业[J]. 1994, 22 (2) : 81~85.
杨洁彬主编.乳酸菌－生物学基础及应用[M].北京：中国轻工业出版社.1998, 100~108.
Atanley, E. , Gilliand , H. S. Kim. Effect of viable starter culture bacteria in yogurt on lactose utilization in humans [J]. Dairy Science . 1984 ,67 : 1~6.
14 Jiang ,T , A. Mustapha , D.Savaiano . Improvement of lactose digestion in human by ingestion of unfermented milk containing Bifidobacterium logum [J] . Dairy Science . 1996 , 79 : 750~757.
15 Anley ,E . ,Gilliland , H . S . Kim . Effect of viable starter culture bacteria in yogurt on lactose utilization in humans [J] . Dairy Science . 1984 , 67 : 1~6.
16 Kim, G. S , S. E.Gilliland .Lactobacillus acidophilus as a dietary adjunct for milk to acid lactose digestion on humans [J]. Dairy Science . 1983 , 66 : 959~966.
Rajagopal , S. N. ,W. E. Sandine. Associative Growth and Proteolysis of streptococcus thermophilus and lactobacillus buglarricus in skim milk [J]. Dairy Science . 1990 , 73 : 894-899.
Tamine, A. Y. ,R. K. Robinson . Yogurt Science and technology [M]. Woodhead Publishing Limited. 2000 : 389~423.
郭本恒. 酸乳 [M]. 北京:化学工业出版社. 2004, 278~365.
谢继志,范立冬,赵平. 液态乳制品科学与技术[M]. 北京 : 国轻工业出版社. 1998, 258~268.
Patrick F .Fox , Timothy P . Guinee , Timothy M . Cogan . Foundamentals of cheese science [M]. Aspen Publishers . 2000:236~423.
郭本恒. 干酪 [M]. 北京:化学工业出版社. 2004, 323~354.
23 Alm L . Effect of fermentation on B- Vitamin content of milk Sweden [J]. Dairy Science . 1982 , 65 : 353~359.
24 Breslaw ,E .S., D. H. Kleyn . In vitro digestibility of protein in yogurt at various stages of processing [J] .Journal of Food Science . 1973 , 38 : 41016~1021.
25 Friend, B. A , K. M. Shahanii .Nutritional and therapeutic aspects of lactobacilli [J]. Journal of Applied Nutrition . 1984 , 36 : 125~153.
Danele Atlan ,Patrichanlo. Isolation and characterization of Aminopeptidase-deficient lactobacillus mutants[J]. Applied Environmental Microbiology. 1989, 7 : 1717~1723.
Fred ,A., Exterkate .Location of peptidase outside and inside the membrance of streptococcus cremoris[J]. Applied Environmental Microbiology. 1984, 1 : 177~183.
Anala, G. , Abraham , Gracielal , De Antoni ,Maria C Anion . Protelytic activity of lactobacillus bulgaricus grown in milk[J] . Dairy Science. 1993 , 76 : 1498~1505.
Argyle, P.J.,Q. E. Mathison, R. C. Chandad. Producion of cell-bound proteinase by lactobacillus bulgaricus and its location in the bacterial cell[J]. Applied Bacteriology. 1976, 41 : 175~184.
Kunio Ohmiya, Yasushi Sato. Hydrolysis of casein by intracellodar proteases from lactic acid bacteria [J]. Agricultural Biology Chemistry. 1978, 42 (1) : 7~10.
Jacques Meyer , Regulajdrd. Purification and characterization of X-proly-dipeptidyl-aminopeptides from lactobacillus lactic and from streptococcus thermophilus[J]. Dariy Science . 1978, 70 (4) : 738~745.
Mckay, K. A. ,Baldwin. Stimulaneous loss of proteinase and lactose-utilizing enzyme activities in streptococcus lactis and reversal of loss by transduction[J]. Applited Microbiology. 1974, 28 (3) : 342~346.
Law, B. A., J. Kolstad. Proteolytic systems in lactic acid bacteria [J]. Antonine Leeuwenhoek. 1983, 49 : 225.
郭本恒. 功能性乳制品. 北京: 中国轻工业出版社. 2001 , 56~74, 263~303.
Renz, U., Z. Puhan . Factors promoting bitterness in yoghurt [J]. Milchwissenschaft. 1975, 30 : 265.
Law,B. A. The accelerated ripening of cheese by use of non-conventional starters and enzymies-A
preliminary assessment [J]. International Dairy Fed . 1979, 108 : 40.
Abd El-Salam, M. H., S. El-Shibiny, N. S. Ahmed. Interrelationships of some determinations in assessment of the ripening of Domiati cheese[J]. Dariy Technology . 1973, 28 : 61.
Arnott, D. R., H. A. Morris, W. B. Combs. Effect of certain chemical factors on the melting quality of process cheese[J]. Dairy Science . 1957, 40 : 957.
Lawab, G. A. ,A. A. Hofi . Testing cheese ripening by rapid chemical techniques[J]. Dariy Science. 1966, 19 : 39.
Vakaleris, D. G., W. V. Price . A rapid spectrophotometric method for measueing cheese ripening [J]. Dariy Science. 1959, 42 : 264.
Fox, P. F., Protelysis during cheese manufacture and ripening [J] Dariy Science . 1989, 1379~1400.
Chandad,R. C., P. J. Argyle, G. E. Mathison. Action of lactobacillus bulgaricus proteinase preparation on milk proteins[J]. Dairy Science. 1982, 65 : 1408~1413.
Lowric, R. J., R. C. Lawrence. A new hyphothesis to accout for the development of bitterness[J]. N. Z. J.Dairy Science Technology. 1972, 7 : 51.
Mills, O. E., T. D. Thomas. Bitterness development in cheddar cheese: Effect of the level of starter proteinase[J]. Nethland Dairy Science Technology. 1980, 15 : 131.
Robert Havennar , Ios H J Huis . Probiotics : a genernal view [M] . Lactic Acid Bacteria . 1994 : 17~25,151~170.
46 廖名龙.几种微生态制剂的介绍.中国微生态杂志[J].1997, 9 (1) : 38
47 布坎南 R E，吉本斯 N E .伯杰氏细菌鉴定手册（第八版）[M].第十六部分革兰氏阳性不生芽孢的杆状细菌.1984 , 797~798,805~806.
宫春波，谢丽源，贺睿非. 酸乳中益生菌活性影响因素的研究概况. 饮料工业[J]. 2003，5 (3) : 32~37.
49 Sreekumar O , Hosono A . Immediate effect of Lactobacillus acidophilus on the intestinal flore and fecal enzymes of rats and the virto inbibition of Escherichiacoli in co- culture [J]. Dairy Science .2000 , 83 : 931~939.
50 Muealidhara, K. S. , G. G. Sheggeby , P. R. Elliker . Effect of feeding lactobacilli on coliform and lactobacillus flor of intestinal tissue and feces from piglets [J]. Journal of Food Protection . 1997, 40: 288~295.
51 Reid, G. Mini-review : the scientific basic for probiotic strains of Lactobacillus [J]. Applied and Enviromental Microbiology . 1999, 65 : 3763~3766.
Kim, G .S ., S. E.Gilliland .Lactobacillus acidophilus as a dietary adjunct for milk to acid lactose digestion on humans [J]. Dairy Science . 1983 , 66 : 959~966.
53 Klaver, F .A. M. , Roelof Van der Meer .The assumed assimilation of cholesterol by lactobacilli and
Bifidobacterium bifidum is due to their bile salt –deconjugation activity [J]. Applied and Enviromental Microbiology. 4993 , 59 : 1120~1124.
54 Shida K , A . Makinok , Morishita .Lacobacillus casei inhibits antigen Induced IgE seretion through regulation of cytokine production in murine splenocyte cultured [J] . International archives of Allergy and Immunology . 1998 , 115 (4) :278~287.
55 Matuzaki T ,R Yamazaki, Hashimotos .The effect of oral feeding of Lacobacillus casei strain shirota on immunoglobulinE production in mice [J] .Dairy Science .1998 , 81 (1) :48~53.
56 Spanhaak S ,R Havenar ,A. G. Schaafsm .The effect of consumption of milk fermented by Lactobacillus casei strain shiroto on the intestinal microflora and immune paramefers in human [J].European Journal of Clinical Nutrition .1998, 52 (12) :899~907.
57 郭本恒. 液态奶 [M]. 北京:化学工业出版社. 2004, 331~335.
58 Meei-yn Lin, Dennis Savaiano. Influence of nonfremented dariy products in humans [J]. Dariy Science. 1991, 74 : 87~95.
Citti, J. E., W. E. Sandine, P. R. Elliker. β-galactosidase of streptococcus lactis[J]. Bacteriology. 1965, 89(4) : 937~942.
轻工部部颁标准：QB/T 1803-93
李宏军、任健、张宗岩. 牛乳的pH值和酸度. 中国乳品工业[J]. 1998, 24(4) : 18~20.
Craig J. Oberg, Bart C . Weimer, Lynn V. Moyes. Proteolytic characterization of lactobacillus belbrueckii ssp bulgaricus strain by the O-Phthalcdialaldenhyde test and amino acid analysis[J]. Dariy Science. 1991, 74 : 398~403.
Wei-Jenlin, Determing β-galactosidase activity of yogurt culture in skim milk [J]. Dairy Science. 1989, 72 : 351~359.
64 Lin, M. Y ., D. Savaiano , S. Harlander .Influence of nonfermented dairy produtcts containing bacterial starter culture on lactose maldigestion in humans [J] . Dairy Science . 1991 , 74 : 87.
65 Simoons F .J. The geographic hypothesis and lactose malabsorption A weighing of the evidence [J]. Digestion Disease Science . 1978 , 23 : 963.
66 Dewit ,O. P. ,Porchart , J.F. Desjeux . Breath hydrogen concentration after lactose ,milk , fresh ,or heat yogurt ingestion by healthy young adults with or without lactose malmbsorption .[J]. Journal Nutrition .1988 , 4 :131.
67 Kolars ,J. C. ,M.. D. Levitt ,M. Aouji , D.A. Savaino Yogurt –an autodigesting source of Lactose [J]. New England Journal Fed . 1984 , 310 : 1.
68 Marteau ,P. B ,Flourie P.,Porchart, J. F. Chastang . Desleux . Effect of the microbial lactose (E.C.3.2.1.23)activity in yogurt on the intestinal absorption of lactose :an study in lactose –deficient humans.[J]. British Journal Nutrition. 1990 , 64(4) : 71.
69 Sandie ,W. E . Roles of Lactobacillus in the intestinal tract [J]. Journal of Food Protection . 1979 , 42 : 259~262.
70 D. O. Noh, S. E. Gilliland .Influence of bile on cellular Integrity and ?-Galactosodase activity of lactobacillus acidophilus [J]. Dairy Science .1993 , 76 : 1253~1259.
71 Noh, D. O. , S. E. Gilliland Influence of Bile on cellular intrgrity and β-galactosidase activity of lactobacillus acidophilus [J]. Dairy Science . 1993, 76 : 1253~1259.
72 Ducasetelle.A, J. Lenoir . Contrition aletude de la flore microbienne du fromage de type saint –paulin [M]. Le Lait . 1969 :489~490,615.
73 Van der Zant W.C.,F.E. Nelson .Protelysis by streptococcus lactics grow in milk with and without controlled pH [J]. Dairy Science .1953 , 36 : 1104.
74 Argyle P. J. ,G. E. Mathson, R. C. Chandan . Prodction of cell –bound proteinase by lactobacillus bulgaricus and its location in the bacterial cell [J]. Applied Bacterial. 1974 , 47 : 175.
75 Ezzat ,.N .M . ,El Soda,.M. J. Desmazeaued , A Ismail . Peptide hydrolases from the thermobacterium group of lactobacill[J]. Milchwissenschaft . 1982 , 37 : 666.
Hull S. E , Studies on milk protein :Colorimetric determintion of the partial hydrolysis of the proteins in milk [J]. Dairy Science . 1947 , 30 : 113.
Thomas,T. D. , O. E. Mills. Proteolytic enzymes of starter bacteria [J]. Nethland Milk Dairy . 1981, 35 : 255.
78 Law, B. A. Peptide utilization by group N streptococci [J]. General Microbiology .1978 , 105 :103.
79 Mac lead ,P. ,D. F. Gordon Jr .peptides as sources of essential amino acid for lactic streptococci .Lactobacillus acidophilus as a dietary adjunct for milk to acid lactose digestion on humans [J]. Dairy Science . 1983 , 66 : 959~966.
80 Iton T. Purification and properties of ?-galactosidase from lactobacillus bulgaricus.[J]. Milchwissenschaft. 1980 , 35 : 593.
Benedi Bioenergetic model of a mixed production fermentation [J]. Biotechnology Bioengineering . 1990 , 130 : 142~ 148.
Kanasaki, M. , S. Brcheny , A. J. Hillier ,G. R. Jago . Effect of temperature on the grow and cicd production of lactic acid bacteria [J]. Daity Technology . 1975, 30 : 142.
Marth ,E H . Microbiological and chemical aspects of cheddar cheese ripening .A review [J] . Dairy Science . 1963 , 46 : 869.
Richardk ，Robinson . Dairy Microbiology Handbook [M]. A John Wiley & Sons . Inc . Publication . 1998 ，494～495 .
Fuller R . Probiotics: an Overview . Human Health : The Contribution of Microorgnisms [M]. 1992:63~73.
骆承庠主编.乳与乳制品工艺学[M] . 北京：农业出版社 , 1992: 39.
顾瑞霞主编.乳与乳制品的生理功能特性[M].北京: 中国轻工业出版社.2000: 294~460.
Bayless ,T . M. , N. S. Rosensweig .A racial diffrernce in incidience of lactose deficiency : A survrey of milk intolerance and lactase deficiency in healthy adult males [J]. American Jouenal Medicine Assay . 1966 , 197 : 968.
Haemmerli ,U .P. ,H .Kistler , R . Amman ,T.Marthaler , G . Semenza , S. Auricchio , A . Prader . Acquired milk intolerance in the adult caused by lactose intestinal lactase activity [J]. American Journal Medicine . 1965 , 38 : 7.
Littman, A ., J .B . Hammond . Diarrhea in adult cause by deficiency in intestinal disaccharidase [J] . Gastroenterology . 1965 . 48 : 237.
Schaafsma, G. ,p . , P . R . Dekker , H . Warrd . Microbial lactase activity and digestion of lactose [J]. Nethessland Milk Dairy . 1998 . 42 : 121.
Norton ,S ., M. D. Rosensweig . Adult human milk intolerance and intestinal lactase deficiency [J]. Dairy Science 1959 , 52 (5) : 585~587.
Dennis ,A, Savaiano . The influence and microbe-containing dairy foods [J]. Dairy Science . 1987,70:397~406.
B.A.费林得. 酸奶菌种选择对风味、抗微生物活性、β-半乳糖苷酶和B族维生素的影响. 中国乳品工业[J]. 1994, 22 (2) : 81~85.
杨洁彬主编.乳酸菌－生物学基础及应用[M].北京：中国轻工业出版社.1998, 100~108.
Atanley, E. , Gilliand , H. S. Kim. Effect of viable starter culture bacteria in yogurt on lactose utilization in humans [J]. Dairy Science . 1984 ,67 : 1~6.
14 Jiang ,T , A. Mustapha , D.Savaiano . Improvement of lactose digestion in human by ingestion of unfermented milk containing Bifidobacterium logum [J] . Dairy Science . 1996 , 79 : 750~757.
15 Anley ,E . ,Gilliland , H . S . Kim . Effect of viable starter culture bacteria in yogurt on lactose utilization in humans [J] . Dairy Science . 1984 , 67 : 1~6.
16 Kim, G. S , S. E.Gilliland .Lactobacillus acidophilus as a dietary adjunct for milk to acid lactose digestion on humans [J]. Dairy Science . 1983 , 66 : 959~966.
Rajagopal , S. N. ,W. E. Sandine. Associative Growth and Proteolysis of streptococcus thermophilus and lactobacillus buglarricus in skim milk [J]. Dairy Science . 1990 , 73 : 894-899.
Tamine, A. Y. ,R. K. Robinson . Yogurt Science and technology [M]. Woodhead Publishing Limited. 2000 : 389~423.
郭本恒. 酸乳 [M]. 北京:化学工业出版社. 2004, 278~365.
谢继志,范立冬,赵平. 液态乳制品科学与技术[M]. 北京 : 国轻工业出版社. 1998, 258~268.
Patrick F .Fox , Timothy P . Guinee , Timothy M . Cogan . Foundamentals of cheese science [M]. Aspen Publishers . 2000:236~423.
郭本恒. 干酪 [M]. 北京:化学工业出版社. 2004, 323~354.
23 Alm L . Effect of fermentation on B- Vitamin content of milk Sweden [J]. Dairy Science . 1982 , 65 : 353~359.
24 Breslaw ,E .S., D. H. Kleyn . In vitro digestibility of protein in yogurt at various stages of processing [J] .Journal of Food Science . 1973 , 38 : 41016~1021.
25 Friend, B. A , K. M. Shahanii .Nutritional and therapeutic aspects of lactobacilli [J]. Journal of Applied Nutrition . 1984 , 36 : 125~153.
Danele Atlan ,Patrichanlo. Isolation and characterization of Aminopeptidase-deficient lactobacillus mutants[J]. Applied Environmental Microbiology. 1989, 7 : 1717~1723.
Fred ,A., Exterkate .Location of peptidase outside and inside the membrance of streptococcus cremoris[J]. Applied Environmental Microbiology. 1984, 1 : 177~183.
Anala, G. , Abraham , Gracielal , De Antoni ,Maria C Anion . Protelytic activity of lactobacillus bulgaricus grown in milk[J] . Dairy Science. 1993 , 76 : 1498~1505.
Argyle, P.J.,Q. E. Mathison, R. C. Chandad. Producion of cell-bound proteinase by lactobacillus bulgaricus and its location in the bacterial cell[J]. Applied Bacteriology. 1976, 41 : 175~184.
Kunio Ohmiya, Yasushi Sato. Hydrolysis of casein by intracellodar proteases from lactic acid bacteria [J]. Agricultural Biology Chemistry. 1978, 42 (1) : 7~10.
Jacques Meyer , Regulajdrd. Purification and characterization of X-proly-dipeptidyl-aminopeptides from lactobacillus lactic and from streptococcus thermophilus[J]. Dariy Science . 1978, 70 (4) : 738~745.
Mckay, K. A. ,Baldwin. Stimulaneous loss of proteinase and lactose-utilizing enzyme activities in streptococcus lactis and reversal of loss by transduction[J]. Applited Microbiology. 1974, 28 (3) : 342~346.
Law, B. A., J. Kolstad. Proteolytic systems in lactic acid bacteria [J]. Antonine Leeuwenhoek. 1983, 49 : 225.
郭本恒. 功能性乳制品. 北京: 中国轻工业出版社. 2001 , 56~74, 263~303.
Renz, U., Z. Puhan . Factors promoting bitterness in yoghurt [J]. Milchwissenschaft. 1975, 30 : 265.
Law,B. A. The accelerated ripening of cheese by use of non-conventional starters and enzymies-A
preliminary assessment [J]. International Dairy Fed . 1979, 108 : 40.
Abd El-Salam, M. H., S. El-Shibiny, N. S. Ahmed. Interrelationships of some determinations in assessment of the ripening of Domiati cheese[J]. Dariy Technology . 1973, 28 : 61.
Arnott, D. R., H. A. Morris, W. B. Combs. Effect of certain chemical factors on the melting quality of process cheese[J]. Dairy Science . 1957, 40 : 957.
Lawab, G. A. ,A. A. Hofi . Testing cheese ripening by rapid chemical techniques[J]. Dariy Science. 1966, 19 : 39.
Vakaleris, D. G., W. V. Price . A rapid spectrophotometric method for measueing cheese ripening [J]. Dariy Science. 1959, 42 : 264.
Fox, P. F., Protelysis during cheese manufacture and ripening [J] Dariy Science . 1989, 1379~1400.
Chandad,R. C., P. J. Argyle, G. E. Mathison. Action of lactobacillus bulgaricus proteinase preparation on milk proteins[J]. Dairy Science. 1982, 65 : 1408~1413.
Lowric, R. J., R. C. Lawrence. A new hyphothesis to accout for the development of bitterness[J]. N. Z. J.Dairy Science Technology. 1972, 7 : 51.
Mills, O. E., T. D. Thomas. Bitterness development in cheddar cheese: Effect of the level of starter proteinase[J]. Nethland Dairy Science Technology. 1980, 15 : 131.
Robert Havennar , Ios H J Huis . Probiotics : a genernal view [M] . Lactic Acid Bacteria . 1994 : 17~25,151~170.
46 廖名龙.几种微生态制剂的介绍.中国微生态杂志[J].1997, 9 (1) : 38
47 布坎南 R E，吉本斯 N E .伯杰氏细菌鉴定手册（第八版）[M].第十六部分革兰氏阳性不生芽孢的杆状细菌.1984 , 797~798,805~806.
宫春波，谢丽源，贺睿非. 酸乳中益生菌活性影响因素的研究概况. 饮料工业[J]. 2003，5 (3) : 32~37.
49 Sreekumar O , Hosono A . Immediate effect of Lactobacillus acidophilus on the intestinal flore and fecal enzymes of rats and the virto inbibition of Escherichiacoli in co- culture [J]. Dairy Science .2000 , 83 : 931~939.
50 Muealidhara, K. S. , G. G. Sheggeby , P. R. Elliker . Effect of feeding lactobacilli on coliform and lactobacillus flor of intestinal tissue and feces from piglets [J]. Journal of Food Protection . 1997, 40: 288~295.
51 Reid, G. Mini-review : the scientific basic for probiotic strains of Lactobacillus [J]. Applied and Enviromental Microbiology . 1999, 65 : 3763~3766.
Kim, G .S ., S. E.Gilliland .Lactobacillus acidophilus as a dietary adjunct for milk to acid lactose digestion on humans [J]. Dairy Science . 1983 , 66 : 959~966.
53 Klaver, F .A. M. , Roelof Van der Meer .The assumed assimilation of cholesterol by lactobacilli and
Bifidobacterium bifidum is due to their bile salt –deconjugation activity [J]. Applied and Enviromental Microbiology. 4993 , 59 : 1120~1124.
54 Shida K , A . Makinok , Morishita .Lacobacillus casei inhibits antigen Induced IgE seretion through regulation of cytokine production in murine splenocyte cultured [J] . International archives of Allergy and Immunology . 1998 , 115 (4) :278~287.
55 Matuzaki T ,R Yamazaki, Hashimotos .The effect of oral feeding of Lacobacillus casei strain shirota on immunoglobulinE production in mice [J] .Dairy Science .1998 , 81 (1) :48~53.
56 Spanhaak S ,R Havenar ,A. G. Schaafsm .The effect of consumption of milk fermented by Lactobacillus casei strain shiroto on the intestinal microflora and immune paramefers in human [J].European Journal of Clinical Nutrition .1998, 52 (12) :899~907.
57 郭本恒. 液态奶 [M]. 北京:化学工业出版社. 2004, 331~335.
58 Meei-yn Lin, Dennis Savaiano. Influence of nonfremented dariy products in humans [J]. Dariy Science. 1991, 74 : 87~95.
Citti, J. E., W. E. Sandine, P. R. Elliker. β-galactosidase of streptococcus lactis[J]. Bacteriology. 1965, 89(4) : 937~942.
轻工部部颁标准：QB/T 1803-93
李宏军、任健、张宗岩. 牛乳的pH值和酸度. 中国乳品工业[J]. 1998, 24(4) : 18~20.
Craig J. Oberg, Bart C . Weimer, Lynn V. Moyes. Proteolytic characterization of lactobacillus belbrueckii ssp bulgaricus strain by the O-Phthalcdialaldenhyde test and amino acid analysis[J]. Dariy Science. 1991, 74 : 398~403.
Wei-Jenlin, Determing β-galactosidase activity of yogurt culture in skim milk [J]. Dairy Science. 1989, 72 : 351~359.
64 Lin, M. Y ., D. Savaiano , S. Harlander .Influence of nonfermented dairy produtcts containing bacterial starter culture on lactose maldigestion in humans [J] . Dairy Science . 1991 , 74 : 87.
65 Simoons F .J. The geographic hypothesis and lactose malabsorption A weighing of the evidence [J]. Digestion Disease Science . 1978 , 23 : 963.
66 Dewit ,O. P. ,Porchart , J.F. Desjeux . Breath hydrogen concentration after lactose ,milk , fresh ,or heat yogurt ingestion by healthy young adults with or without lactose malmbsorption .[J]. Journal Nutrition .1988 , 4 :131.
67 Kolars ,J. C. ,M.. D. Levitt ,M. Aouji , D.A. Savaino Yogurt –an autodigesting source of Lactose [J]. New England Journal Fed . 1984 , 310 : 1.
68 Marteau ,P. B ,Flourie P.,Porchart, J. F. Chastang . Desleux . Effect of the microbial lactose (E.C.3.2.1.23)activity in yogurt on the intestinal absorption of lactose :an study in lactose –deficient humans.[J]. British Journal Nutrition. 1990 , 64(4) : 71.
69 Sandie ,W. E . Roles of Lactobacillus in the intestinal tract [J]. Journal of Food Protection . 1979 , 42 : 259~262.
70 D. O. Noh, S. E. Gilliland .Influence of bile on cellular Integrity and ?-Galactosodase activity of lactobacillus acidophilus [J]. Dairy Science .1993 , 76 : 1253~1259.
71 Noh, D. O. , S. E. Gilliland Influence of Bile on cellular intrgrity and β-galactosidase activity of lactobacillus acidophilus [J]. Dairy Science . 1993, 76 : 1253~1259.
72 Ducasetelle.A, J. Lenoir . Contrition aletude de la flore microbienne du fromage de type saint –paulin [M]. Le Lait . 1969 :489~490,615.
73 Van der Zant W.C.,F.E. Nelson .Protelysis by streptococcus lactics grow in milk with and without controlled pH [J]. Dairy Science .1953 , 36 : 1104.
74 Argyle P. J. ,G. E. Mathson, R. C. Chandan . Prodction of cell –bound proteinase by lactobacillus bulgaricus and its location in the bacterial cell [J]. Applied Bacterial. 1974 , 47 : 175.
75 Ezzat ,.N .M . ,El Soda,.M. J. Desmazeaued , A Ismail . Peptide hydrolases from the thermobacterium group of lactobacill[J]. Milchwissenschaft . 1982 , 37 : 666.
Hull S. E , Studies on milk protein :Colorimetric determintion of the partial hydrolysis of the proteins in milk [J]. Dairy Science . 1947 , 30 : 113.
Thomas,T. D. , O. E. Mills. Proteolytic enzymes of starter bacteria [J]. Nethland Milk Dairy . 1981, 35 : 255.
78 Law, B. A. Peptide utilization by group N streptococci [J]. General Microbiology .1978 , 105 :103.
79 Mac lead ,P. ,D. F. Gordon Jr .peptides as sources of essential amino acid for lactic streptococci .Lactobacillus acidophilus as a dietary adjunct for milk to acid lactose digestion on humans [J]. Dairy Science . 1983 , 66 : 959~966.
80 Iton T. Purification and properties of ?-galactosidase from lactobacillus bulgaricus.[J]. Milchwissenschaft. 1980 , 35 : 593.
Benedi Bioenergetic model of a mixed production fermentation [J]. Biotechnology Bioengineering . 1990 , 130 : 142~ 148.
Kanasaki, M. , S. Brcheny , A. J. Hillier ,G. R. Jago . Effect of temperature on the grow and cicd production of lactic acid bacteria [J]. Daity Technology . 1975, 30 : 142.
Marth ,E H . Microbiological and chemical aspects of cheddar cheese ripening .A review [J] . Dairy Science . 1963 , 46 : 869.
Richardk ，Robinson . Dairy Microbiology Handbook [M]. A John Wiley & Sons . Inc . Publication . 1998 ，494～495 .