产邻苯二酚工程菌的构建及发酵条件的优化  

The Formation of Recombinant Strain Producing Catechol and the Optimization of Fermentation Conditions

作　　者：张宗武[1] 梁璇[1] 张敏[1] 李俊芳[1] 武波[1] 

机构地区：[1]广西大学生命科学与技术学院微生物及植物遗传工程教育部重点实验室广西亚热带生物资源保护利用重点实验室,南宁530005

出　　处：《中国生物工程杂志》2006年第9期67-71,共5页China Biotechnology

基　　金：国家"863"计划资助项目(2003AA214040);广西科技攻关资助项目(0537012-A)

摘　　要：从实验室保存的一株能高效降解苯胺的不动杆菌中克隆到完整的苯胺双加氧酶基因簇,序列分析表明该基因簇包含6个完整的ORF,全序列与已报道的不动杆菌YAA的苯胺双加氧酶基因簇在氨基酸水平上有较高的同源性。将该基因簇连接于pLAFR6载体,电转化至E.coli,构建了该基因簇的工程菌。发酵条件优化表明,在苯胺浓度0.5mg/ml,采用pH7.0的LB培养基,E.coliDH5α为宿主菌,于37℃培养,接种量为3%的条件下,邻苯二酚产量在20h达到0.546mg/ml,底物分子水平转化率可达92.4%。A complete aniline dioxygenase gene cluster cloned from an Acinetobacter sp. strain, which could utilize aniline as the sole carbon, nitrogen and energy, was sequenced. Sequence analysis showed that the gene cluster had six intact ORFs, and the whole sequence had high similarity with that of Acinetobacter sp. YAA at amino acid level. A recombinant strain was formed with the gene cluster 1/gated to vector pLAFR6 and transferred to E. coll. After optimizing the fermentation conditions of this strain for producing catechol, LB was confirmed as the final medium, pH7.0, aniline concentration 0.5mg/ml, E. coli DHSαas the host, incubation temperature 37℃, amount of inoculum 3%. Under above conditions, the yield of catechol could get to 0.546mg/ml, and the converting rate of substrate at molecule level could get to 92.4%.

关 键 词：苯胺 苯胺双加氧酶基因簇 邻苯二酚 

分 类 号：TQ93[化学工程]