Enantioselective synthesis of pure (R,R)-2,3-butanediol in Escherichia coli with stereospecific secondary alcohol dehydrogenases.

We characterized the activity and stereospecificity of four secondary alcohol dehydrogenases (sADHs) towards acetoin reduction and constructed synthetic pathways in E. coli to produce enantiomerically pure (R,R)-2,3-butanediol (2,3-BDO) from glucose with a titer of 6.1 g/L (enantio purity >99%), and yield of 0.31 g product/g glucose (62% of theoretical maximum).

[1]  K. Prather,et al.  Metabolic Engineering of Escherichia coli for Enhanced Production of (R)- and (S)-3-Hydroxybutyrate , 2009, Applied and Environmental Microbiology.

[2]  Andreas Liese,et al.  Biocatalytic ketone reduction—a powerful tool for the production of chiral alcohols—part I: processes with isolated enzymes , 2007, Applied Microbiology and Biotechnology.

[3]  S. Ui,et al.  Discovery of a new mechanism of 2,3-butanediol stereoisomer formation in Bacillus cereus YUF-4 , 1998 .

[4]  M. Syu.  Biological production of 2,3-butanediol , 2001, Applied Microbiology and Biotechnology.

[5]  T. Ohtsuki,et al.  Production of l‐2,3‐butanediol by a new pathway constructed in Escherichia coli , 2004, Letters in applied microbiology.

[6]  J. C. Liao,et al.  Engineered Synthetic Pathway for Isopropanol Production in Escherichia coli , 2007, Applied and Environmental Microbiology.

[7]  Kevin M. Smith,et al.  Metabolic engineering of Escherichia coli for 1-butanol production. , 2008, Metabolic engineering.

[8]  V. Fülöp,et al.  Purification, crystallization and preliminary X-ray crystallographic studies on acetolactate decarboxylase. , 2003, Acta crystallographica. Section D, Biological crystallography.

[9]  W. Nicholson.  The Bacillus subtilis ydjL (bdhA) Gene Encodes Acetoin Reductase/2,3-Butanediol Dehydrogenase , 2008, Applied and Environmental Microbiology.

[10]  T. Kudo,et al.  Molecular generation of an Escherichia coli strain producing only the meso-isomer of 2,3-butanediol , 1997 .

[11]  Andreas Liese,et al.  Biocatalytic ketone reduction—a powerful tool for the production of chiral alcohols—part II: whole-cell reductions , 2007, Applied Microbiology and Biotechnology.

[12]  Anton Glieder,et al.  Yeast cell factories for fine chemical and API production , 2008, Microbial cell factories.