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Applied Microbiology and Biotechnology

, Volume 68, Issue 2, pp 228–236 | Cite as

Single-gene knockout of a novel regulatory element confers ethionine resistance and elevates methionine production in Corynebacterium glutamicum

  • Jörg MampelEmail author
  • Hartwig Schröder
  • Stefan Haefner
  • Uwe Sauer
Applied Genetics and Molecular Biotechnology

Abstract

Despite the availability of genome data and recent advances in methionine regulation in Corynebacterium glutamicum, sulfur metabolism and its underlying molecular mechanisms are still poorly characterized in this organism. Here, we describe the identification of an ORF coding for a putative regulatory protein that controls the expression of genes involved in sulfur reduction dependent on extracellular methionine levels. C. glutamicum was randomly mutagenized by transposon mutagenesis and 7,000 mutants were screened for rapid growth on agar plates containing the methionine antimetabolite d,l-ethionine. In all obtained mutants, the site of insertion was located in the ORF NCgl2640 of unknown function that has several homologues in other bacteria. All mutants exhibited similar ethionine resistance and this phenotype could be transferred to another strain by the defined deletion of the NCgl2640 gene. Moreover, inactivation of NCgl2640 resulted in significantly increased methionine production. Using promoter lacZ-fusions of genes involved in sulfur metabolism, we demonstrated the relief of l-methionine repression in the NCgl2640 mutant for cysteine synthase, o-acetylhomoserine sulfhydrolase (metY) and sulfite reductase. Complementation of the mutant strain with plasmid-borne NCgl2640 restored the wild-type phenotype for metY and sulfite reductase.

Keywords

Methionine Corynebacterium Glutamicum Sulfur Metabolism Ethionine Sulfite Reductase 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

We thank L. Eggeling and G. Leblon for kindly providing strain C. glutamicum ATCC14752 and plasmid pCGL0040, respectively.

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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Jörg Mampel
    • 1
    Email author
  • Hartwig Schröder
    • 2
  • Stefan Haefner
    • 2
  • Uwe Sauer
    • 1
  1. 1.Institute of BiotechnologySwiss Federal Institute of Technology (ETH)ZurichSwitzerland
  2. 2.BASF AktiengesellschaftLudwigshafenGermany

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