乳酸菌菌株安全性的研究及优良酸奶发酵剂的筛选
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摘要
乳酸菌是一群能利用碳水化合物产生大量乳酸的细菌的统称。主要有乳杆菌属(如嗜酸乳杆菌、干酪乳杆菌、鼠李糖乳杆菌)、双歧杆菌属(如长双歧杆菌、短双歧杆菌、两歧双歧杆菌、青春双歧杆菌)、链球菌属(如嗜热链球菌)、乳球菌属(如乳酸乳球菌)研究表明,以上菌种均可作为益生菌,具有生理活性,对人体有益。
乳酸菌菌株的药敏性评价是目前国际上益生乳酸菌安全性评价的首要内容,然而用于乳酸菌药敏性测试的相关国际通引标准还未出台。本文就药敏试验使用的培养基进行了探讨,并从中选取了合适的商品化培养基——RCA用于乳酸菌的药敏性测试。选取的培养基适合目前市场上常用益生菌的生长,经方法优化后可得到满意的结果,为今后制定针对乳酸菌的抗菌药物敏感性试验执行标准提供了一定依据。随后采用纸片扩散法对分属于4个属的9株乳酸菌进行了19类51种抗生素的敏感性测定,较全面地了解了这些菌株的药敏谱。
酸奶和乳酸菌是密不可分的,酸奶由于乳酸菌的发酵作用,在营养成分得到改善的同时,也发挥了乳酸菌益生的功效,从而倍受人们青睐。搅拌型酸奶在储存过程中以后酸化及质构等品质问题尤为突出。酸奶发酵剂的优劣与酸奶质量的好坏有着密切关系。本文通过多步筛选,得到了4组发酵剂配方。其发酵的酸奶在28 d货架期内品质都能得到保证且无需添加增稠剂以改善质构缺陷。此外,为了快速指针酸奶货架期后酸化程度的强弱,15组28 d储存期实验与加速实验所得的pH值被用于进行比较,结果显示,加速试验中pH稳定时的值作为判断后酸化程度强弱的指标是可行且快速的。
Lactic Acid Bacteria (LAB) produce lactic acid as the major metabolic end-product of carbohydrate fermentation. The major genera that comprise the LAB are Lactobacillus (such as Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus rhamnosus), Bifidobacterium (such as Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium bifidum, Bifidobacterium adolesentis), Streptococcus (such as Streptococcus thermophilus) and Lactococcus (such as Lactococcus lactis). Researchs show they are beneficial to human body as probiotics.
The evaluation of antibiotic susceptibility of LAB is a major part of the safety of probiotic foods. However, there is no international standard method of the susceptibility testing of LAB till now. We investigated 6 culture media and selected a proper commercial one from them. We optimized the agar disc diffusion method and assayed 9 strains of LAB encompassing the genera Lactobacillus, Bifidobacterium, Streptococcus and Lactococcus for susceptibility to 51 antibiotics on RCA agar. The optimized method proved to be an applicable technique for antibiotic susceptibility testing of LAB. This contributes to the establishment of a standardized procedure for antibiotic susceptibility testing of LAB.
Yoghurt is a dairy product produced by bacterial fermentation of milk. It is inseparable with LAB which make yoghurt rich in nutrition and do good to gastrointestinal tract of human being. Stirred yoghurt is faced with problems such as post-acidification and texture deficiency. Quality of starter culture determines the quality of yoghurt during the shelf life. We selected 4 starter formulas through layers of screening. The quality of yoghurt used with these 4 starter formulas could be assured. The acidification capability was weak and texture deficiency was improved without additional stabilizers during the shelf life of 28 d. In addition, we chose the strains of weak acidification capability by Accelerated Shelf Life Testing. We changed the yoghurt storage temperature to 43 "C (the same as the fermentation temperature) and observed the sample pH till the pH reached stable. This method proved to be effective.
乳酸菌菌株的药敏性评价是目前国际上益生乳酸菌安全性评价的首要内容,然而用于乳酸菌药敏性测试的相关国际通引标准还未出台。本文就药敏试验使用的培养基进行了探讨,并从中选取了合适的商品化培养基——RCA用于乳酸菌的药敏性测试。选取的培养基适合目前市场上常用益生菌的生长,经方法优化后可得到满意的结果,为今后制定针对乳酸菌的抗菌药物敏感性试验执行标准提供了一定依据。随后采用纸片扩散法对分属于4个属的9株乳酸菌进行了19类51种抗生素的敏感性测定,较全面地了解了这些菌株的药敏谱。
酸奶和乳酸菌是密不可分的,酸奶由于乳酸菌的发酵作用,在营养成分得到改善的同时,也发挥了乳酸菌益生的功效,从而倍受人们青睐。搅拌型酸奶在储存过程中以后酸化及质构等品质问题尤为突出。酸奶发酵剂的优劣与酸奶质量的好坏有着密切关系。本文通过多步筛选,得到了4组发酵剂配方。其发酵的酸奶在28 d货架期内品质都能得到保证且无需添加增稠剂以改善质构缺陷。此外,为了快速指针酸奶货架期后酸化程度的强弱,15组28 d储存期实验与加速实验所得的pH值被用于进行比较,结果显示,加速试验中pH稳定时的值作为判断后酸化程度强弱的指标是可行且快速的。
Lactic Acid Bacteria (LAB) produce lactic acid as the major metabolic end-product of carbohydrate fermentation. The major genera that comprise the LAB are Lactobacillus (such as Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus rhamnosus), Bifidobacterium (such as Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium bifidum, Bifidobacterium adolesentis), Streptococcus (such as Streptococcus thermophilus) and Lactococcus (such as Lactococcus lactis). Researchs show they are beneficial to human body as probiotics.
The evaluation of antibiotic susceptibility of LAB is a major part of the safety of probiotic foods. However, there is no international standard method of the susceptibility testing of LAB till now. We investigated 6 culture media and selected a proper commercial one from them. We optimized the agar disc diffusion method and assayed 9 strains of LAB encompassing the genera Lactobacillus, Bifidobacterium, Streptococcus and Lactococcus for susceptibility to 51 antibiotics on RCA agar. The optimized method proved to be an applicable technique for antibiotic susceptibility testing of LAB. This contributes to the establishment of a standardized procedure for antibiotic susceptibility testing of LAB.
Yoghurt is a dairy product produced by bacterial fermentation of milk. It is inseparable with LAB which make yoghurt rich in nutrition and do good to gastrointestinal tract of human being. Stirred yoghurt is faced with problems such as post-acidification and texture deficiency. Quality of starter culture determines the quality of yoghurt during the shelf life. We selected 4 starter formulas through layers of screening. The quality of yoghurt used with these 4 starter formulas could be assured. The acidification capability was weak and texture deficiency was improved without additional stabilizers during the shelf life of 28 d. In addition, we chose the strains of weak acidification capability by Accelerated Shelf Life Testing. We changed the yoghurt storage temperature to 43 "C (the same as the fermentation temperature) and observed the sample pH till the pH reached stable. This method proved to be effective.
引文
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[2]金世琳.乳酸菌的科学与技术[J].中国乳品工业,1998,26(2)：14-16.
[3]张刚.乳酸细菌——基础、技术和应用[M].北京：化学工业出版社,2007.
[4]东秀珠.常见细菌系统鉴定手册[M].北京：科学出版社,2001.
[5]顾瑞霞,谢继志.乳酸菌与人体保健[M].北京：科学出版社,1995.
[6]郭本恒.现代乳品技工技术丛书——酸奶[M].北京：化学工业出版社,2003.
[7]GB 19302-2003酸乳卫生标准
[8]许本发.酸奶和乳酸菌饮料加工[M].北京：中国轻工业出版社,1994.
[9]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,2001,17(3)：43-46.
[10]丛玉婷,高学军.DVS乳酸菌种在我国的应用及其开发研制[J].中国乳品工业,2004,32(10)：27-29.
[11]Vinderola CG, Mocchiutti P, Reinheimer JA. Interaction among with lactic acid starter and probiotic bacteria used for fermented dairy products[J]. J Dairy Sci,2002,85: 721-729.
[12]徐成勇,吴昊,郑思聪,苏永红,诸葛健,郭本恒.弱后酸化酸奶发酵剂的筛选[J].中国乳品工业,2007,35(3)：12-16.
[13]霍贵成.乳酸菌的研究与应用[M].北京：中国轻工业出版社,2007.
[14]Alonso L, Fraga MJ. Simple and rapid analysis for quantitation of the most important volatile flavor compounds in yogurt by headspace gas chromatography mass spectrometry [J]. J. Chromato. Sci.,2001,39(7):297-300.
[15]Adams MR. Safety of industrial lactic and bacteria[J]. J Biotechnic,1999,68:171-178.
[16]胡妹敏,杨秀茹,谢永民.益生菌BB-46及LA-5在酸奶中的应用[A].中国乳制品工业协会第十二次年会中国乳制品工业协会第十二次年会论文集[C].2006.89-92.
[17]崔岸.益生菌种的应用和市场趋势[J].广州食品工业科技,2003,19(增刊)：71-72.
[18]张英春,张兰威,李妍.嗜酸乳杆菌和婴儿双歧杆菌对肠道致病菌的抑制性[J].东北农业大学学报,2004,35(2)：208-211.
[19]Lin MY, Yen CL. Antioxidative ability of lactic acid bacteria[J]. J Agric Food Chem, 1999,47:1460-1466.
[20]Kullisaar T, Zilmer M, Mikelsaar M, et al. Two antioxidative lactobacilli strains as promising probiotics[J]. Int J Food Microbiol,2002,72:215-224.
[21]Kullisaar T, Songisepp E, Mikelsaar M, et al. Antioxidative probiotic fermented goats'
milk decreases oxidative stress-mediated atherogenicity in human subjects[J]. Br J Nutr, 2003,90:449-456.
[22]高学云,芒来.发酵乳中功能性乳酸菌的研究进展[J].内蒙古科技与经济,2007,(09s)：39-40.
[23]Lin SY, Ayres JW, Winkler W Jr, et al. Lactobacillus effects on cholesterol:in vitro and in vivo results[J].1989,72:2885-2889.
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