Microbial production of <Emphasis Type="Italic">cis</Emphasis>-1,2-dihydroxy-cyclohexa-3,5-diene-1-carboxylate by genetically modified <Emphasis Type="Italic">Pseudomonas putida</Emphasis> |
| |
Authors: | Shang-Yu Sun Xin Zhang Qin Zhou Jin-Chun Chen Guo-Qiang Chen |
| |
Institution: | (1) Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing, 100084, China;(2) Multidisciplinary Research Center, Shantou University, Shantou, 515063, China |
| |
Abstract: | Arene cis-diols are interesting chemicals because of their chiral structures and great potentials in industrial synthesis of useful
chiral chemical products. Pseudomonas putida KT2442 was genetically modified to transform benzoic acid (benzoate) to 1,2-dihydroxy-cyclohexa-3,5-diene-1-carboxylic acid
(DHCDC) or named benzoate cis-diol. BenD gene encoding cis-diol dehydrogenase was deleted to generate a mutant named P. putida KTSY01. Genes benABC encoding benzoate dioxygenase were cloned into plasmid pSYM01 and overexpressed in P. putida KTSY01. The recombinant bacteria P. putida KTSY01 (pSYM01) showed strong ability to transform benzoate to DHCDC. DHCDC of 2.3 g/L was obtained with a yield of 73% after
24 h of cultivation in shake flasks incubated under optimized growth conditions. Transformation of benzoate carried out in
a 6-L fermentor using a benzoate fed-batch process produced over 17 g/L DHCDC after 48 h of fermentation. The average DHCDC
production rate was 0.356 g L−1 h−1. DHCDC purified from the fermentation broth showed a purity of more than 95%, and its chemical structure was confirmed by
nuclear magnetic resonance. |
| |
Keywords: | Pseudomonas putida Benzoate dioxygenase cis-diol dehydrogenase Biotransformation cis-1 2-dihydroxy-cyclohexa-3 5-diene-1-carboxylic acid DHCDC |
本文献已被 PubMed SpringerLink 等数据库收录! |
|