Premium
Optimization of expression of dha T gene encoding 1,3‐propanediol oxidoreductase from Klebsiella pneumoniae in Escherichia coli using the methods of uniform design and regression analysis
Author(s) -
Cao Yang,
Xia Qirong,
Fang Baishan
Publication year - 2006
Publication title -
journal of chemical technology and biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.64
H-Index - 117
eISSN - 1097-4660
pISSN - 0268-2575
DOI - 10.1002/jctb.1360
Subject(s) - inducer , lac operon , 1,3 propanediol , yield (engineering) , oxidoreductase , isopropyl , chemistry , gene , gene expression , biochemistry , fermentation , enzyme , organic chemistry , physics , thermodynamics
This paper is concerned with the optimization of expression conditions of 1,3‐propanediol oxidoreductase (PDOR) using the uniform design (UD) method and regression analysis. The concentration of the inducer isopropyl‐β‐ D ‐thiogalactopyranoside (IPTG), the induction temperature and the induction time, which were identified as three crucial factors, were optimized in 50 mL cultivation tubes containing 5 mL LB culture. The optimal expression conditions for enhanced PDOR activity were found to be an IPTG concentration of 0.1 mmol l −1 , an induction temperature of 21.2 °C and an induction time of 5 h. Subsequent experimental trials confirmed the validity of the model. These optimal expression conditions in the cultivation tube culture led to a PDOR activity of 9.25 U mL −1 , which was considerably higher than those obtained in the optimizing experimental trials. Copyright © 2005 Society of Chemical Industry