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费氏链霉菌中子辐射诱变菌株对新霉素的生物转化
引用本文:马林,达娃,刘运,史冠莹,杨茜,杨从军,杨凌云,张新刚,陶科,侯太平. 费氏链霉菌中子辐射诱变菌株对新霉素的生物转化[J]. 微生物学通报, 2010, 37(9): 1347-1355
作者姓名:马林  达娃  刘运  史冠莹  杨茜  杨从军  杨凌云  张新刚  陶科  侯太平
作者单位:1. 四川大学生命科学学院,四川,成都,610064;四川大学生物资源与生态环境教育部重点实验室,四川,成都,610064
2. 西藏高原生物研究所,西藏,拉萨,850001
基金项目:国家自然科学基金项目(No. 30800034)
摘    要:对新霉素产生菌费氏链霉菌进行中子辐射诱变和突变株筛选, 获得不产新霉素的突变株。以突变株为转化菌种, 以新霉素为底物, 对转化发酵液进行高效液相色谱分析, 研究了不同转化条件对新霉素转化的影响。结果表明, 底物浓度、底物添加时间、底物添加方式、接种量、培养基装量、转化时间、碳源、氮源、pH、温度对新霉素转化具有不同程度的影响。以转化条件优化参数进行转化大量培养, 转化液经4步离子交换层析进行分离纯化, 薄层层析检测纯化样品为单一斑点。采用薄层生物自显影对获得的4个转化产物分离样品做生物活性检测, 发现4个样品对金黄色葡萄球菌和姜青枯假单孢杆菌都具有抑制活性, 1个样品对大白菜软腐样品具有明显的抑制活性。

关 键 词:新霉素   生物转化   辐射诱变   费氏链霉菌   离子交换层析   抑菌活性

Biotransformation of Neomycin by Mutational Strain of Streptomyces fradiae Mutagenized with Neutron Radiation
MA Lin,DA W,LIU Yun,SHI Guan-Ying,YANG Qian,YANG Cong-Jun,YANG Ling-Yun,ZHANG Xin-Gang,TAO Ke and HOU Tai-Ping. Biotransformation of Neomycin by Mutational Strain of Streptomyces fradiae Mutagenized with Neutron Radiation[J]. Microbiology China, 2010, 37(9): 1347-1355
Authors:MA Lin  DA W  LIU Yun  SHI Guan-Ying  YANG Qian  YANG Cong-Jun  YANG Ling-Yun  ZHANG Xin-Gang  TAO Ke  HOU Tai-Ping
Affiliation:1. College of Life Sciences, Sichuan University, Chengdu, Sichuan 610064, China; 2. Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, Chengdu, Sichuan 610064, China;3. Tibet Institute of Plateau Biology, Lhasa, Tibet, 850000, China;1. College of Life Sciences, Sichuan University, Chengdu, Sichuan 610064, China; 2. Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, Chengdu, Sichuan 610064, China;1. College of Life Sciences, Sichuan University, Chengdu, Sichuan 610064, China; 2. Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, Chengdu, Sichuan 610064, China;1. College of Life Sciences, Sichuan University, Chengdu, Sichuan 610064, China; 2. Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, Chengdu, Sichuan 610064, China;1. College of Life Sciences, Sichuan University, Chengdu, Sichuan 610064, China; 2. Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, Chengdu, Sichuan 610064, China;1. College of Life Sciences, Sichuan University, Chengdu, Sichuan 610064, China; 2. Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, Chengdu, Sichuan 610064, China;1. College of Life Sciences, Sichuan University, Chengdu, Sichuan 610064, China; 2. Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, Chengdu, Sichuan 610064, China;1. College of Life Sciences, Sichuan University, Chengdu, Sichuan 610064, China; 2. Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, Chengdu, Sichuan 610064, China;1. College of Life Sciences, Sichuan University, Chengdu, Sichuan 610064, China; 2. Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, Chengdu, Sichuan 610064, China
Abstract:A mutant of neomycin-producer Streptomyces fradiae was obtained from cultures mutagenized with neutron radiation. Utilizing the mutational strain which can not produce neomycin in normal culture, the biotransformation of neomycin was studied at different conversion conditions. The analysis results of fermentation broth by high performance liquid chromatography showed that the culture conditions such as the concentration of substrate, adding time of substrate, adding method of substrate, inoculation amount, the volume of medium, transformation time, carbon sources, nitrogen sources, pH of media and temperature had different influence on neomycin transformation. The transformation products of neomycin were isolated from a large scale fermentation broth by ion exchange chromatography and identified by thin layer chromatography. The antibacterial activity of four purified samples against Staphylococcus aureus, Erwinia carotovora and Pseudomonas solanacarum were tested by bioautographic analysis. All four samples showed obvious inhibitory activity against Staphylococcus aureus and Pseudomonas solanacarum, but only one sample had inhibitory effect against Erwinia carotovora.
Keywords:Neomycin   Biotransformation   Irradiation mutagenesis   Streptomyces fradiae   Ion exchange chromatography   Antibacterial activity
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