产二甲苯单加氧酶菌株的筛选、鉴定及转化2,5-二甲基吡嗪条件的优化  

Isolation and Identification of A Xylene Monooxygenase-Producing Strain and Optimization of the Transformation Conditions of 2,5-Dimethyl Pyrazine

作　　者：刘丽娟[1] 薛亚平[1] 郑裕国[1] 

机构地区：[1]浙江工业大学生物与环境工程学院,浙江杭州310014

出　　处：《化学与生物工程》2012年第9期19-25,共7页Chemistry & Bioengineering

基　　金：国家科技部973计划项目(2007CB714306)

摘　　要：从土壤中筛选得到一株可以选择性氧化2,5-二甲基吡嗪生成5-甲基吡嗪-2-羧酸的产单加氧酶菌株ZJB-LLJ,经过生理生化鉴定及16S rDNA序列分析确定为恶臭假单胞菌。并对其转化2,5-二甲基吡嗪产5-甲基吡嗪-2-羧酸的条件进行了优化,结果表明,在500mL摇瓶中装液100mL、温度为30℃、pH值为7～8、以对二甲苯为唯一碳源、诱导培养12h后添加底物的条件下,产物积累量最大;采用分批流加的方式积累产物较快,转化528h后5-甲基吡嗪-2-羧酸浓度达20.41g·L-1,产率达到75.6%。A xylene monooxygenase-producing strain ZJB-LLJ which can selectively oxidize 2,5-dimethyl pyrazine to 5-methylpyrazine-2-carboxylic acid was isolated from soil. According to its physiological and biochemical characters and 16S rDNA sequence analysis,the strain was identified taxonomically as Pseudomonas putida. The reaction conditions of batch biotransformation for the production of 5-methylpyrazine-2-carboxylic acid from 2,5-dimethyl pyrazine were optimized. The highest production was obtained when Pseudomonas putida was cultured using p-xylene as the sole carbon source in 100 mL medium（pH value of 7-8） in a 500 mL flask at 30 ℃ and 2,5-dimethyl pyrazine was added after 12 h. In fed-batch biotransformation, 2,5-dimethyl pyrazine was added to the reaction system continuously,5-methylpyrazine-2-carboxylic acid concentration could reach 20.41 g . L-1 with a yield of 75.6% at 528 h.

关 键 词：5-甲基吡嗪-2-羧酸 2 5-二甲基吡嗪 二甲苯单加氧酶 筛选鉴定 

分 类 号：Q814.1[生物学—生物工程]