The regulation of 2,3-butanediol synthesis in Klebsiella pneumoniae as revealed by gene over-expressions and metabolic flux analysis

Mingshou Lu, Changhun Park, Soojin Lee, Borim Kim, Min-Kyu Oh, Youngsoon Um, Jungwook Kim, Jinwon Lee

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

A variety of microorganism species are able naturally to produce 2,3-butanediol (2,3-BDO), although only a few of them are suitable for consideration as having potential for mass production purposes. Klebsiella pneumoniae (K. pneumoniae) is one such strain which has been widely studied and used industrially to produce 2,3-BDO. In the central carbon metabolism of K. pneumoniae, the 2,3-BDO synthesis pathway is dominated by three essential enzymes, namely acetolactate decarboxylase, acetolactate synthase, and butanediol dehydrogenase, which are encoded by the budA, budB, and budC genes, respectively. The mechanisms of the three enzymes have been characterized with regard to their function and roles in 2,3-BDO synthesis and cell growth (Blomqvist et al. in J Bacteriol 175(5):1392-1404, 1993), while a few studies have focused on the cooperative mechanisms of the three enzymes and their mutual interactions. Therefore, the K. pneumoniae KCTC2242::ΔwabG wild-type strain was utilized to reconstruct seven new mutants by single, double, and triple overexpression of the three enzymes key to this study. Subsequently, continuous cultures were performed to obtain steady-state metabolism in the organisms and experimental data were analyzed by metabolic flux analysis (MFA) to determine the regulation mechanisms. The MFA results showed that the seven overexpressed mutants all exhibited enhanced 2,3-BDO production, and the strain overexpressing the budBA gene produced the highest yield. While the enzyme encoded by the budA gene produced branched-chain amino acids which were favorable for cell growth, the budB gene enzyme rapidly enhanced the conversion of acetolactate to acetoin in an oxygen-dependent manner, and the budC gene enzyme catalyzed the reversible conversion of acetoin to 2,3-BDO and regulated the intracellular NAD+/NADH balance.

Original languageEnglish
Pages (from-to)343-353
Number of pages11
JournalBioprocess and Biosystems Engineering
Volume37
Issue number3
DOIs
Publication statusPublished - 2014 Jan 1

Fingerprint

Metabolic Flux Analysis
Klebsiella pneumoniae
Enzymes
Genes
Fluxes
Gene Expression
Acetoin
acetolactate decarboxylase
butanediol dehydrogenase
Cell growth
Metabolism
NAD
Acetolactate Synthase
Branched Chain Amino Acids
Growth
2,3-butylene glycol
Microorganisms
Amino acids
Carbon
Oxygen

Keywords

  • 2,3-butanediol
  • Acetolactate decarboxylase
  • Acetolactate synthase
  • Butantediol dehydrogenase
  • Metabolic flux analysis

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering

Cite this

The regulation of 2,3-butanediol synthesis in Klebsiella pneumoniae as revealed by gene over-expressions and metabolic flux analysis. / Lu, Mingshou; Park, Changhun; Lee, Soojin; Kim, Borim; Oh, Min-Kyu; Um, Youngsoon; Kim, Jungwook; Lee, Jinwon.

In: Bioprocess and Biosystems Engineering, Vol. 37, No. 3, 01.01.2014, p. 343-353.

Research output: Contribution to journalArticle

Lu, Mingshou ; Park, Changhun ; Lee, Soojin ; Kim, Borim ; Oh, Min-Kyu ; Um, Youngsoon ; Kim, Jungwook ; Lee, Jinwon. / The regulation of 2,3-butanediol synthesis in Klebsiella pneumoniae as revealed by gene over-expressions and metabolic flux analysis. In: Bioprocess and Biosystems Engineering. 2014 ; Vol. 37, No. 3. pp. 343-353.
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