Contracted but effective: production of enantiopure 2,3-butanediol by thermophilic and GRAS Bacillus licheniformis

The production of chemicals with chiral groups through chemical methods is attractive but difficult to perform. Optically active 2,3-butanediol (2,3-BD) can provide two chiral groups and has special applications in asymmetric synthesis. However, the chemical production of 2,3-BD from nonrenewable resources is expensive to perform and results in a mixture of three stereoisomeric forms. Enantiopure 2,3-BD production through green biotechnological routes is thus rather desirable. In this work, we introduced a “SAME” (Screening, Analysis, Mutation, and Evaluation) process to achieve efficient microbes for chiral chemicals’ production using 2,3-BD as a target molecule. Bacillus licheniformis MW3, a GRAS and thermophilic strain, was firstly selected as a promising host. The mechanism of stereoisomer formation of 2,3-BD in B. licheniformis MW3 was then investigated. Two stereospecific 2,3-BD dehydrogenases (BDHs), 2R,3R-BDH (encoded by gdh) and meso-BDH (encoded by budC), were identified to be responsible for the production of (2R,3R)-2,3-BD and meso-2,3-BD. Two metabolically engineered strains, B. licheniformis MW3 (ΔbudC) and B. licheniformis MW3 (Δgdh), were constructed for the production of (2R,3R)-2,3-BD and meso-2,3-BD, respectively. After 42 h of fermentation, 123.7 g L−1 (2R,3R)-2,3-BD was synthesized using B. licheniformis MW3 (ΔbudC); while 90.1 g L−1meso-2,3-BD was synthesized after 32 h using B. licheniformis MW3 (Δgdh). Because of their high levels of production and biosecurity, these engineered B. licheniformis strains might have potential in the commercial production of enantiopure (2R,3R)-2,3-BD and meso-2,3-BD. The contracted but effective “SAME” process might serve as an alternative method for other chiral chemicals’ production.

[1]  Sang Yup Lee,et al.  Recent advances in microbial production of fuels and chemicals using tools and strategies of systems metabolic engineering. , 2015, Biotechnology advances.

[2]  M. Arnaud,et al.  New Vector for Efficient Allelic Replacement in Naturally Nontransformable, Low-GC-Content, Gram-Positive Bacteria , 2004, Applied and Environmental Microbiology.

[3]  M. Faulstich,et al.  Enhanced fed-batch fermentation of 2,3-butanediol by Paenibacillus polymyxa DSM 365. , 2012, Bioresource technology.

[4]  H. Liesegang,et al.  Size unlimited markerless deletions by a transconjugative plasmid-system in Bacillus licheniformis. , 2013, Journal of biotechnology.

[5]  R. Sheldon Green and sustainable manufacture of chemicals from biomass: state of the art , 2014 .

[6]  S. Atsumi,et al.  Combinatorial optimization of cyanobacterial 2,3-butanediol production. , 2014, Metabolic engineering.

[7]  J. Lu,et al.  Thermophilic fermentation of acetoin and 2,3-butanediol by a novel Geobacillus strain , 2012, Biotechnology for Biofuels.

[8]  Yajun Yan,et al.  Enantioselective synthesis of pure (R,R)-2,3-butanediol in Escherichia coli with stereospecific secondary alcohol dehydrogenases. , 2009, Organic & biomolecular chemistry.

[9]  Charlotte K. Williams,et al.  The Path Forward for Biofuels and Biomaterials , 2006, Science.

[10]  Yong-Su Jin,et al.  Production of 2,3-butanediol from xylose by engineered Saccharomyces cerevisiae. , 2014, Journal of biotechnology.

[11]  J. Keasling Synthetic biology and the development of tools for metabolic engineering. , 2012, Metabolic engineering.

[12]  Huimin Zhao,et al.  Metabolic engineering of a Saccharomyces cerevisiae strain capable of simultaneously utilizing glucose and galactose to produce enantiopure (2R,3R)-butanediol. , 2014, Metabolic engineering.

[13]  Hong Xu,et al.  A 2,3‐butanediol dehydrogenase from Paenibacillus polymyxa ZJ‐9 for mainly producing R,R‐2,3‐butanediol: Purification, characterization and cloning , 2013, Journal of basic microbiology.

[14]  P. Ouyang,et al.  Microbial 2,3-butanediol production: a state-of-the-art review. , 2011, Biotechnology advances.

[15]  S Dusko Ehrlich,et al.  Complete genome sequence of the industrial bacterium Bacillus licheniformis and comparisons with closely related Bacillus species , 2004, Genome Biology.

[16]  F. Meinhardt,et al.  Generation of readily transformable Bacillus licheniformis mutants , 2008, Applied Microbiology and Biotechnology.

[17]  K. Jantama,et al.  Efficient reduction of the formation of by-products and improvement of production yield of 2,3-butanediol by a combined deletion of alcohol dehydrogenase, acetate kinase-phosphotransacetylase, and lactate dehydrogenase genes in metabolically engineered Klebsiella oxytoca in mineral salts medium. , 2015, Metabolic engineering.

[18]  W. Grajek,et al.  Biotechnological production of 2,3-butanediol--current state and prospects. , 2009, Biotechnology advances.

[19]  Avelino Corma,et al.  Conversion of biomass platform molecules into fuel additives and liquid hydrocarbon fuels , 2014 .

[20]  D. Wei,et al.  Mechanism of 2,3-butanediol stereoisomer formation in Klebsiella pneumoniae , 2014, Applied Microbiology and Biotechnology.

[21]  Zhiyou Wen,et al.  Deletion of meso-2,3-butanediol dehydrogenase gene budC for enhanced D-2,3-butanediol production in Bacillus licheniformis , 2014, Biotechnology for Biofuels.

[22]  Cuiqing Ma,et al.  Systematic metabolic engineering of Escherichia coli for high-yield production of fuel bio-chemical 2,3-butanediol. , 2014, Metabolic engineering.

[23]  Shangtian Yang,et al.  Production of 2,3‐butanediol from glucose by GRAS microorganism Bacillus amyloliquefaciens , 2011, Journal of basic microbiology.

[24]  Gerald A. Tuskan,et al.  Lignin Valorization: Improving Lignin Processing in the Biorefinery , 2014, Science.

[25]  Cuiqing Ma,et al.  Metabolic engineering of Enterobacter cloacae for high-yield production of enantiopure (2R,3R)-2,3-butanediol from lignocellulose-derived sugars. , 2015, Metabolic engineering.

[26]  X. Parés,et al.  Characterization of a (2R,3R)-2,3-Butanediol Dehydrogenase as theSaccharomyces cerevisiae YAL060W Gene Product , 2000, The Journal of Biological Chemistry.

[27]  Yu Wang,et al.  Glycerol Dehydrogenase Plays a Dual Role in Glycerol Metabolism and 2,3-Butanediol Formation in Klebsiella pneumoniae * , 2014, The Journal of Biological Chemistry.

[28]  Sang Yup Lee,et al.  Biorefineries for the production of top building block chemicals and their derivatives. , 2015, Metabolic engineering.

[29]  T. Ohtsuki,et al.  Purification and Characterization of L-2,3-Butanediol Dehydrogenase of Brevibacterium saccharolyticum C-1012 Expressed in Escherichia coli , 2001, Bioscience, biotechnology, and biochemistry.

[30]  Klaas J Hellingwerf,et al.  Synthesis of 2,3-butanediol by Synechocystis sp. PCC6803 via heterologous expression of a catabolic pathway from lactic acid- and enterobacteria. , 2013, Metabolic engineering.

[31]  Cuiqing Ma,et al.  A Novel Whole-Cell Biocatalyst with NAD+ Regeneration for Production of Chiral Chemicals , 2010, PloS one.

[32]  Cuiqing Ma,et al.  Efficient 2,3-Butanediol Production from Cassava Powder by a Crop-Biomass-Utilizer, Enterobacter cloacae subsp. dissolvens SDM , 2012, PloS one.

[33]  Xiao-Jun Ji,et al.  Constructing a synthetic metabolic pathway in Escherichia coli to produce the enantiomerically pure (R, R)-2,3-butanediol. , 2015, Biotechnology and bioengineering.

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

[35]  G. G. Stokes "J." , 1890, The New Yale Book of Quotations.

[36]  Ping Xu,et al.  Metabolic engineering of Escherichia coli for production of (2S,3S)-butane-2,3-diol from glucose , 2015, Biotechnology for Biofuels.

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

[38]  Cuiqing Ma,et al.  Biocatalytic production of (2S,3S)-2,3-butanediol from diacetyl using whole cells of engineered Escherichia coli. , 2012, Bioresource technology.

[39]  S. Lee,et al.  Systems strategies for developing industrial microbial strains , 2015, Nature Biotechnology.

[40]  D. M. Alonso,et al.  Targeted chemical upgrading of lignocellulosic biomass to platform molecules , 2014 .

[41]  S. Atsumi,et al.  A carbon sink pathway increases carbon productivity in cyanobacteria. , 2015, Metabolic engineering.

[42]  Cuiqing Ma,et al.  A newly isolated Bacillus licheniformis strain thermophilically produces 2,3-butanediol, a platform and fuel bio-chemical , 2013, Biotechnology for Biofuels.

[43]  J. Chu,et al.  Microbial production of 2,3-butanediol by a surfactant (serrawettin)-deficient mutant of Serratia marcescens H30 , 2010, Journal of Industrial Microbiology & Biotechnology.

[44]  R. Sheldon,et al.  Comparison of the sustainability metrics of the petrochemical and biomass-based routes to methionine , 2015 .

[45]  Cuiqing Ma,et al.  Enhanced 2,3-butanediol production by Klebsiella pneumoniae SDM , 2009, Applied Microbiology and Biotechnology.

[46]  J. van der Oost,et al.  d-2,3-Butanediol Production Due to Heterologous Expression of an Acetoin Reductase in Clostridium acetobutylicum , 2011, Applied and Environmental Microbiology.

[47]  Li Sha,et al.  BIOTECHNOLOGICALLY RELEVANT ENZYMES AND PROTEINS A new NAD(H)-dependent meso-2,3-butanediol dehydrogenase from an industrially potential strain Serratia marcescens H30 , 2014 .

[48]  Jibin Sun,et al.  Novel (2R,3R)-2,3-Butanediol Dehydrogenase from Potential Industrial Strain Paenibacillus polymyxa ATCC 12321 , 2011, Applied and Environmental Microbiology.

[49]  Andrew G. Glen,et al.  APPL , 2001 .

[50]  Cuiqing Ma,et al.  Genome Sequence of Enterobacter cloacae subsp. dissolvens SDM, an Efficient Biomass-Utilizing Producer of Platform Chemical 2,3-Butanediol , 2012, Journal of bacteriology.

[51]  Shujing Sun,et al.  Cloning, expression and characterization of glycerol dehydrogenase involved in 2,3-butanediol formation in Serratia marcescens H30 , 2014, Journal of Industrial Microbiology & Biotechnology.

[52]  Joseph J. Bozell,et al.  Technology development for the production of biobased products from biorefinery carbohydrates—the US Department of Energy’s “Top 10” revisited , 2010 .

[53]  L. Gouveia,et al.  Green metrics evaluation of isoprene production by microalgae and bacteria , 2013 .

[54]  Ke-Ke Cheng,et al.  Improved 2,3-butanediol production from corncob acid hydrolysate by fed-batch fermentation using Klebsiella oxytoca , 2010 .

[55]  Rachit Jain,et al.  Inhibition of acetate accumulation leads to enhanced production of (R,R)-2,3-butanediol from glycerol in Escherichia coli , 2012, Journal of Industrial Microbiology & Biotechnology.

[56]  Rainer Merkl,et al.  The Complete Genome Sequence of Bacillus licheniformis DSM13, an Organism with Great Industrial Potential , 2004, Journal of Molecular Microbiology and Biotechnology.

[57]  T. Kudo,et al.  Cloning, expression and nucleotide sequence of the L-2,3-butanediol dehydrogenase gene from Brevibacterium saccharolyticum C-1012 , 1998 .