title: A non-pathogenic and optically high concentrated (R,R)-2,3-butanediol biosynthesizing Klebsiella strain.
abstract: The objective of this work was to construct a non-pathogenic Klebsiella pneumonia strain that can produce optically high concentrated (R,R)-2,3-BDO. A K. pneumonia mutant lacking the pathogenic factor was used as the host strain. In order to construct a K. pneumonia strain that would biosynthesize high concentrated (R,R)-2,3-BDO, gene deletion and over-expression methods were combined; firstly, the 2,3-BDO dehydrogenase (budC) gene was deleted to re-direct utilization of the carbon source to (R,R)-2,3-BDO biosynthesis; secondly, the two glycerol dehydrogenase (GDH) enzymes in K. pneumonia (DhaD and GldA) were over-expressed to maximize (R,R)-2,3-BDO biosynthesis; and thirdly, the lactate dehydrogenase (ldhA) gene was deleted to minimize the accumulation of lactate. SGSB112, a non-pathogenic strain of K. pneumonia that can produce optically high concentrated (R,R)-2,3-BDO, was constructed as above. Approximately 36% of the carbon source was converted to (R,R)-2,3-BDO by SGSB112, achieving a production of 61gL(-1) (R,R)-2,3-BDO in a fed-batch fermentation. On the other hand, meso-2,3-BDO was produced 1.4gL(-1) and (S,S)-2,3-BDO was not detected. This study provides an insight into 2,3-BDO biosynthesis in K. pneumonia and demonstrates the achievement of high-yield production of optically high concentrated (R,R)-2,3-BDO through constructing a strain by genetic modification and metabolic engineering.
authors.affiliation: Department of Chemical and Biomolecular Engineering, Sogang University, 35 Baekbeom-ro, Mapo-gu, Seoul 121-742, South Korea.
authors.forename: Soojin
authors.initials: S
authors.lastname: Lee
authors.affiliation: Department of Chemical and Biomolecular Engineering, Sogang University, 35 Baekbeom-ro, Mapo-gu, Seoul 121-742, South Korea.
authors.forename: Borim
authors.initials: B
authors.lastname: Kim
authors.affiliation: Department of Chemical and Biomolecular Engineering, Sogang University, 35 Baekbeom-ro, Mapo-gu, Seoul 121-742, South Korea.
authors.forename: Jeongmo
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Chemical and Biomolecular Engineering, Sogang University, 35 Baekbeom-ro, Mapo-gu, Seoul 121-742, South Korea.
authors.forename: Daun
authors.initials: D
authors.lastname: Jeong
authors.affiliation: Department of Chemical and Biomolecular Engineering, Sogang University, 35 Baekbeom-ro, Mapo-gu, Seoul 121-742, South Korea.
authors.forename: Soohyun
authors.initials: S
authors.lastname: Park
authors.affiliation: Department of Chemical and Biomolecular Engineering, Sogang University, 35 Baekbeom-ro, Mapo-gu, Seoul 121-742, South Korea. Electronic address: jinwonlee1@gmail.com.
authors.forename: Jinwon
authors.initials: J
authors.lastname: Lee
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D061249:Batch Cell Culture Techniques / Q000379:methods; D002072:Butylene Glycols / Q000378:metabolism*; D017353:Gene Deletion; D024861:Genetic Enhancement / Q000379:methods*; D007709:Klebsiella / Q000235:genetics / Q000378:metabolism*; D060847:Metabolic Engineering / Q000379:methods
chemical_list: D001426:Bacterial Proteins; D002072:Butylene Glycols; C026978:2,3-butylene glycol
keywords: (R,R)-2,3-butanediol; 2,3-Butanediol stereoisomers