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Constructing a synthetic metabolic pathway in Escherichia coli to produce the enantiomerically pure (R, R)‐2,3‐butanediol
Author(s) -
Ji XiaoJun,
Liu LuGang,
Shen MengQiu,
Nie ZhiKui,
Tong YingJia,
Huang He
Publication year - 2015
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.25512
Subject(s) - 2,3 butanediol , escherichia coli , 1,4 butanediol , fermentation , biosynthesis , butanediol , lac operon , chemistry , biochemistry , recombinant dna , isopropyl , metabolic pathway , metabolic engineering , enzyme , stereochemistry , organic chemistry , catalysis , gene
ABSTRACT Enantiomerically pure (R, R)‐2,3‐butanediol has unique applications due to its special chiral group and spatial configuration. Currently, its chemical production route has many limitations. In addition, no native microorganisms can accumulate (R, R)‐2,3‐butanediol with an enantio‐purity over 99%. Herein, we constructed a synthetic metabolic pathway for enantiomerically pure (R, R)‐2,3‐butanediol biosynthesis in Escherichia coli . The fermentation results suggested that introduction of the synthetic metabolic pathway redistributed the carbon fluxes to the neutral (R, R)‐2,3‐butanediol, and thus protected the strain against the acetic acid inhibition. Additionally, it showed that the traditionally used isopropyl beta‐D‐thiogalactoside (IPTG) induction displayed negative effect on (R, R)‐2,3‐butanediol biosynthesis in the recombinant E. coli , which was probably due to the protein burden. With no IPTG addition, the (R, R)‐2,3‐butanediol concentration reached 115 g/L by fed‐batch culturing of the recombinant E. coli , with an enantio‐purity over 99%, which is suitable for the pilot‐scale production. Biotechnol. Bioeng. 2015;112: 1056–1059. © 2014 Wiley Periodicals, Inc.