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Cited by 175Bacterial Genetics and BiotechnologyFungal and yeast genetics researchMicrobial Metabolic Engineering and Bioproduction

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Synthesis of the osmoprotectant glycine betaine in Bacillus subtilis: characterization of the gbsAB genes.

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Boch, J, Kempf, B, Schmid, R, Bremer, E·1996·Journal of Bacteriology
DOI 10.1128/jb.178.17.5121-5129.1996PMC PMC178307
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Abstract

Synthesis of the osmoprotectant glycine betaine from the exogenously provided precursor choline or glycine betaine aldehyde confers considerable osmotic stress tolerance to Bacillus subtilis in high-osmolarity media. Using an Escherichia coli mutant (betBA) defective in the glycine betaine synthesis enzymes, we cloned by functional complementation the genes that are required for the synthesis of the osmoprotectant glycine betaine in B. subtilis. The DNA sequence of a 4.1-kb segment from the cloned chromosomal B. subtilis DNA was established, and two genes (gbsA and gbsB) whose products were essential for glycine betaine biosynthesis and osmoprotection were identified. The gbsA and gbsB genes are transcribed in the same direction, are separated by a short intergenic region, and are likely to form an operon. The deduced gbsA gene product exhibits strong sequence identity with members of a superfamily of specialized and nonspecialized aldehyde dehydrogenases. This superfamily comprises glycine betaine aldehyde dehydrogenases from bacteria and plants with known involvement in the cellular adaptation to high-osmolarity stress and drought. The deduced gbsB gene product shows significant similarity to the family of type III alcohol dehydrogenases. B. subtilis mutants with defects in the chromosomal gbsAB genes were constructed by marker replacement, and the growth properties of these mutant strains in high-osmolarity medium were analyzed. Deletion of the gbsAB genes destroyed the choline-glycine betaine synthesis pathway and abolished the ability of B. subtilis to deal effectively with high-osmolarity stress in choline- or glycine betaine aldehyde-containing medium. Uptake of radiolabelled choline was unaltered in the gbsAB mutant strain. The continued intracellular accumulation of choline or glycine betaine aldehyde in a strain lacking the glycine betaine-biosynthetic enzymes strongly interfered with the growth of B. subtilis, even in medium of moderate osmolarity. A single transcription initiation site for gbsAB was detected by high-resolution primer extension analysis. gbsAB transcription was initiated from a promoter with close homology to sigma A-dependent promoters and was stimulated by the presence of choline in the growth medium.

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Metadata

Authors
Boch, J., Kempf, B., Schmid, R., & Bremer, E.
Affiliations
Max Planck Society (DE)
ORCID
Venue
Journal of Bacteriology
Publisher
American Society for Microbiology
Published
1996-09
Volume
178
Issue
17
Pages
5121-5129
ISSN
  • 0021-9193 (ISSN-L)
  • 1098-5530
DOI
10.1128/jb.178.17.5121-5129.1996
PMC
PMC178307
OpenAlex
W2017036247
Cited by
175
References
0
Citations by year
2026: 5 · 2025: 10 · 2024: 8 · 2023: 7 · 2022: 12 · 2021: 7 · 2020: 10 · 2019: 5 · 2018: 7 · 2017: 7 · 2016: 4 · 2015: 2 · 2014: 8 · 2013: 8 · 2012: 3
FWCI
4.41
Type
journal article
Language
en
Open access
Licence
journals.asm.org/non-commercial-tdm-license (tdm)
Locations
  • Journal of Bacteriology (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
Deutsche Forschungsgemeinschaft
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
Keywords
glycine betaine biosynthesis, osmoprotectants, osmotic stress tolerance, Bacillus subtilis, choline, aldehyde dehydrogenase, operon, transcription initiation, functional complementation, drought adaptation
MeSH terms
Alcohol Dehydrogenase, Aldehyde Dehydrogenase, Amino Acid Sequence, Bacillus subtilis, Bacterial Proteins, Base Sequence, Betaine, Choline, Cloning, Molecular, DNA, Bacterial, Escherichia coli, Molecular Sequence Data, Osmolar Concentration, Plasmids, Gene Deletion, Sequence Analysis, DNA

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