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应用双向荧光差异凝胶技术解析干酪乳杆菌Lactobacillus casei Zhang驯化菌株耐酸胁迫机制
引用本文:张娟,吴重德,张梦汝,堵国成,陈坚.应用双向荧光差异凝胶技术解析干酪乳杆菌Lactobacillus casei Zhang驯化菌株耐酸胁迫机制[J].工业微生物,2012,42(5):62-68.
作者姓名:张娟  吴重德  张梦汝  堵国成  陈坚
作者单位:1. 食品科学与技术国家重点实验室 无锡214122;江南大学生物工程学院 无锡214122
2. 四川大学轻纺与食品学院 成都610065
3. 糖化学与生物技术教育部重点实验室 无锡214122;江南大学生物工程学院 无锡214122
4. 粮食发酵工艺与技术国家工程实验室 无锡214122;江南大学生物工程学院 无锡214122
基金项目:国家高技术研究发展计划(863项目,2011AA100901);国家自然科学基金重点项目(20836003);国家自然科学基金(30900013).
摘    要:以干酪乳杆菌Lactobacillus casei Zhang为出发菌株,通过适应性进化获得了干酪乳杆菌酸胁迫抗性驯化菌株.对细胞内微环境的检测发现,驯化菌株在酸胁迫过程中能够维持较高的磷酸烯醇式丙酮酸-糖转移酶系统活力,并具有较高的H+ -ATPase活性以及胞内ATP浓度.蛋白质组学分析结果表明,酸胁迫引发了细胞蛋白表达的变化,与原始菌株相比,驯化菌株保持了更高的代谢活性;同时,驯化菌株通过大量诱导应激蛋白如分子伴侣GroEL、GrpE,冷/热应激蛋白CspC、DnaK等维持了细胞的生理活性,有效提高了细胞对酸胁迫的抵御能力.本研究为进一步揭示酸胁迫下乳酸菌细胞的生理应答机制,探寻促进乳酸菌酸胁迫性能提升的最优策略,进而改善其在生产中的应用性能提供了可借鉴的思路.

关 键 词:干酪乳杆菌  酸胁迫  双向荧光差异凝胶电泳  驯化

Mechanism of Lactobacillus casei Zhang domesticated strain against acid stress by two-dimensional fluorescence difference in gel electrophoresis analysis
ZHANG Juan , WU Chong-de , ZHANG Meng-ru , DU Guo-cheng , CHEN Jian.Mechanism of Lactobacillus casei Zhang domesticated strain against acid stress by two-dimensional fluorescence difference in gel electrophoresis analysis[J].Industrial Microbiology,2012,42(5):62-68.
Authors:ZHANG Juan  WU Chong-de  ZHANG Meng-ru  DU Guo-cheng  CHEN Jian
Institution:1. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China; 2. The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; 3. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China; 4. School of Bioteehnology, Jiangnan University, Wuxi 214122, China; 5. College of Light Industry, Textile and Food Engineering, Siehuan University, Chengdu 610065, China)
Abstract:The domesticated strain of Lactobacillus casei Zhang with the adaptive evolution. The intracellular microenvironment analysis outstanding acid stress resistance was obtained after found that the domesticated strain can maintain a higher phosphoenolpyruvate-sugar transfer enzyme activity in the acid stress process, and showed higher H+ -ATPase activity and intracellular ATP concentration. Proteomics analysis investigated that the acid treatment led to expression changes of cell protein. Compared with the wild strain, the domesticated strain maintained higher metabolic activity Meanwhile, the domesticated strain kept its physiological activity by a large number of induced stress proteins such as molecular chaperones GroEL, GrpE, and the cold / heat stress protein CspC / DnaK, which effectively improved the acid stress resistance. This study revealed the physiology of lactic acid bacteria cells under acid stress further, and explored the optimal strategy to promote the performance of lactic acid bacteria against acid stress, thereby provided a novel way to enhance their performance on industrial application.
Keywords:Lactobacillus casei  acid stress  two-dimensional fluorescence difference in gel electrophoresis  domestication
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