Delftia tsuruhatensis  AD9苯胺双加氧酶基因的过量表达及多功能降解工程菌的构建

›› 2008, Vol. 10 ›› Issue (4): 95-99.

Delftia tsuruhatensis AD9苯胺双加氧酶基因的过量表达及多功能降解工程菌的构建

耿立召梁泉峰张维平淑珍陆伟林敏陈明

中国农业科学院生物技术研究所,北京100081

收稿日期:2008-04-09 修回日期:2008-06-02 出版日期:2008-08-15 发布日期:2009-07-29
通讯作者: 陈明，研究员，主要从事环境微生物的分子生物学研究。Tel：010-68975038；E-mail：chenmingbio@hotmail．com
作者简介:耿立召|博士研究生|主要从事环境微生物基因工程研究。Tel：010-68919861；E-mail：luckyglz@163．com。
基金资助:
国家自然科学基金项目（30470047,30770076）资助.

Over-expression of Aniline Dioxygenase Gene in Delftia tsuruhatensis |AD9 and Construction of Engineering Strain of |Multi-substrate Utilization

GENG Li-zhao, LIANG Quan-feng, ZHANG Wei, PING Shu-zhen, LU Wei, LIN Min, CHEN Ming

Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081|China

Received:2008-04-09 Revised:2008-06-02 Online:2008-08-15 Published:2009-07-29

摘要/Abstract

摘要：

Delftia tsuruhatensis AD9和Acinetobacter calcoaceticus PHEA-2分别能利用苯胺和苯酚作为唯一碳源生长，降解的第一个中间产物均为邻苯二酚，随后裂解为参与三羧酸循环的中间产物。通过PCR方法克隆到苯胺高效降解菌AD9的苯胺双加氧酶基因，并构建表达苯胺双加氧酶的广宿主质粒载体pVD，通过三亲接合，导入到AD9和PHEA-2中。对两种重组菌中苯胺双氧酶基因的表达及苯胺降解特性的分析结果表明，增加苯胺双加氧酶基因的拷贝数可以提高野生型AD9的苯胺降解速率，同时该基因的表达使PHEA-2菌获得苯瞎隆觎能寸1．

关键词: 苯胺双加氧酶 Delftia tsuruhatensis AD9 Acinetobacter calcoaceticus PHEA-2

Abstract:

Delftia tsuruhatensis AID and Acinetobacter calcoaceticus PHEA-2 can utilize aniline and phenol as the sole carbon source, respectively. Catechol is the first mid-metabolites through the degradation of aromatic compounds, which is degrade by certain enzymes to TCA-cycle intermediates. A complete aniline dioxygenase gene from AID strain was cloned into a broad host range plasmid, the resulting plasmid pVD was introduced by triparental mating into wildtype strain AD9 and a phenol-degrading bacterium, A. calcoaceticus PHEA-2, respectively. The over-expression of aniline dioxygenase gene enhanced the aniline degradation of AD9 strain and conferred the capable of aniline utilization to the PHEA-2 strain.

Key words: aniline dioxygenase, Delftia tsuruhatensis AD9, Acinetobacter calcoaceticus PHEA-2

中图分类号:

Q933

耿立召,梁泉峰,张维,平淑珍,陆伟,林敏,陈明. Delftia tsuruhatensis AD9苯胺双加氧酶基因的过量表达及多功能降解工程菌的构建[J]. , 2008, 10(4): 95-99.

GENG Li-zhao, LIANG Quan-feng, ZHANG Wei, PING Shu-zhen, LU Wei, LIN Min, CHEN M. Over-expression of Aniline Dioxygenase Gene in Delftia tsuruhatensis |AD9 and Construction of Engineering Strain of |Multi-substrate Utilization[J]. , 2008, 10(4): 95-99.