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Cited by 14Microbial Metabolic Engineering and BioproductionBacterial Genetics and BiotechnologyMicrobial metabolism and enzyme function

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Enhanced production of branched-chain amino acids by Gluconacetobacter europaeus with a specific regional deletion in a leucine responsive regulator

Akasaka, Naoki, Ishii, Yuri, Hidese, Ryota, Sakoda, Hisao, Fujiwara, Shinsuke·2014·Journal of Bioscience and Bioengineering
DOI 10.1016/j.jbiosc.2014.05.024
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Abstract

Vinegar with increased amounts of branched-chain amino acids (BCAAs; valine, leucine and isoleucine) is favorable for human health as BCAAs decrease diet-induced obesity and hyperglycemia. To construct Gluconacetobacter europaeus which produces BCAAs, leucine responsive regulator (GeLrp) is focused and two Gelrp mutants were constructed. Wild-type KGMA0119 didn't produce significant amount of valine (0.13 mM) and leucine (0 mM) and strain KGMA7110 which lacks complete Gelrp accumulated valine (0.48 mM) and leucine (0.11 mM) but showed impaired growth, and it was fully restored in the presence of essential amino acids. Strain KGMA7203 was then constructed with a nonsense mutation at codon Trp132 in the Gelrp, which leads a specific deletion at an estimated ligand-sensing region in the C-terminal domain. KGMA7203 produced greater quantities of valine (0.80 mM) and leucine (0.26 mM) and showed the same growth characteristics as KGMA0119. mRNA levels of BCAAs biosynthesis genes (ilvI and ilvC) and probable BCAAs efflux pump (leuE) were determined by quantitative reverse-transcription PCR. Expression rates of ilvI and ilvC in the two Gelrp disruptants were greater than those in KGMA0119. leuE was highly expressed in KGMA7110 only, suggesting that the accumulation in KGMA7110 culture was caused by increased expression of the biosynthesis genes and abnormal enhanced export of amino acids resulting in impaired cell growth. In contrast, KGMA7203 would achieve the high level production through enhanced expression of the biosynthesis genes without enhancing that for the efflux pump. KGMA7203 was considered advantageous for production of vinegar with higher amounts of valine and leucine.

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Metadata

Authors
Akasaka, N., Ishii, Y., Hidese, R., Sakoda, H., & Fujiwara, S.
Affiliations
Kwansei Gakuin University (JP)
ORCID
Venue
Journal of Bioscience and Bioengineering
Publisher
Elsevier BV
Published
2014-12
Volume
118
Issue
6
Pages
607-615
ISSN
  • 1347-4421 (ISSN-L)
  • 1389-1723
DOI
10.1016/j.jbiosc.2014.05.024
OpenAlex
W2092572386
Cited by
14
References
0
Citations by year
2026: 1 · 2025: 1 · 2021: 4 · 2020: 1 · 2019: 2 · 2017: 1 · 2016: 2 · 2015: 2
FWCI
0.63
Type
journal article
Language
en
Open access
closed
Licence
www.elsevier.com/tdm/userlicense/1.0/ (tdm)
Locations
  • Journal of Bioscience and Bioengineering (publishedVersion)
  • PubMed (publishedVersion)
Funders
Japan Society for the Promotion of Science (24310015)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial metabolism and enzyme function
Keywords
branched-chain amino acids, isoleucine, vinegar, gene disruption mutants
MeSH terms
Amino Acids, Branched-Chain, Leucine, RNA, Bacterial, RNA, Messenger, Valine, Gluconacetobacter

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