The Formation of Recombinant Strain Producing Catechol and the Optimization of Fermentation Conditions
- VernacularTitle:产邻苯二酚工程菌的构建及发酵条件的优化
- Author:
Zong-Wu ZHANG
;
Xuan LIANG
;
Min ZHANG
;
Jun-Fang LI
;
Bo WU
;
- Publication Type:Journal Article
- Keywords:
Aniline Aniline dioxygenase gene cluster Catechol
- From:
China Biotechnology
2006;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
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 ligated to vector pLAFR6 and transferred to E.coli. 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 DH5?as 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%.