Laboratory of Applied Stress MicrobiologyLaboratory of Applied Stress Microbiology

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Research Projects

3. Physiological roles and metabolic regulation of cysteine in Escherichia coli and their applications to cysteine fermentation

Cysteine is an important amino acid both biologically and commercially. For achieving cysteine fermentation directly from glucose, we analyze the regulatory mechanisms on its biosynthesis, degradation and transport, based on genomic information and metabolic engineering. Also, we are interested in physiological roles of cysteine and its transporters in terms of redox regulation, and recently proposed that the inducible cysteine/cystine shuttle system plays a pivotal role in oxidative stress tolerance through providing a reducing equivalent to the periplasm in Escherichia coli.

cysteine, transporter, redox regulation (oxidative/reductive stress), sulfur metabolism, the cysteine/cystine shuttle system


  1. Y. Tanaka, K. Yoshikaie, A. Takeuchi, M. Ichikawa, T. Mori, S. Uchino, Y. Sugano, T. Hakoshima, H. Takagi, G. Nonaka and T. Tsukazaki*: Crystal structure of a YeeE/YedE family protein engaged in thiosulfate uptake. Sci. Adv., 6, eaba7637 (2020). DOI: 10.1126/sciadv.aba7637
  2. E. Funahashi*, K. Saiki*, K. Honda, Y. Sugiura, Y. Kawano, I. Ohtsu, D. Watanabe, Y. Wakabayashi, T. Abe, T. Nakanishi, M. Suematsu, and H. Takagi: A finding of thiosulfate pathway for synthesis of organic sulfur compounds in Saccharomyces cerevisiae and an improvement of ethanol production. *These authors contributed equally to this work. J. Biosci. Bioeng., 120, 666-669 (2015).
  3. I. Ohtsu, Y. Kawano, M. Suzuki, S. Morigasaki, K. Saiki, S. Yamazaki, G. Nonaka, and H. Takagi: Uptake of l-cystine via an ABC transporter contributes defense of oxidative stress in the l-cysteine export-dependent manner in Escherichia coliPLoS One, 10(4), e0120619 (2015).
  4. Y. Kawano*, I. Ohtsu*, A. Tamakoshi, M. Shiroyama, A. Tsuruoka, K. Saiki, K. Takumi, G. Nonaka, T. Nakanishi, T. Hishiki, M. Suematsu, and H. Takagi: Involvement of the yciW gene in l-cysteine and l-methionine metabolism in Escherichia coli. *These authors contributed equally to this work. J. Biosci. Bioeng., 119, 310-313 (2015).
  5. Y. Kawano*, I. Ohtsu*, K. Takumi, A. Tamakoshi, G. Nonaka, E. Funahashi, M. Ihara, and H. Takagi: Enhancement of l-cysteine production by disruption of yciW in Escherichia coli. *These authors contributed equally to this work. J. Biosci. Bioeng., 119, 176-179 (2015).
  6. T. Nakatani*, I. Ohtsu*, G.Nonaka, N. Wiriyathanawudhiwong, S. Morigasaki and H. Takagi: Enhancement of thioredoxin/glutaredoxin-mediated L-cysteine synthesis from S-sulfocysteine increases L-cysteine production in Escherichia coli. *These authors contributed equally to this work. Microb. Cell Fact., 11:62 doi:10.1186/1475-2859-11-62.
  7. I. Ohtsu*, N. Wiriyathanawudhiwong*, S. Morigasaki, T. Nakatani, H. Kadokura and H. Takagi: The L-cysteine/L-cystine shuttle system provides reducing equivalents to the periplasm in Escherichia coli. *These authors contributed equally to this work. J. Biol. Chem., 285, 17479-17487 (2010).
  8. N. Wiriyathanawudhiwong*, I. Ohtsu*, Z.-D. Li, H. Mori and H. Takagi: The outer membrane TolC is involved in cysteine tolerance and overproduction in Escherichia coli. *These authors contributed equally to this work. Appl. Microbiol. Biotech., 81, 903-913 (2009).
  9. M. Wada and H. Takagi: Metabolic pathways and biotechnological production of L-cysteine. Appl. Microbiol. Biotech., 73, 48-54 (2006).
  10. S. Yamada, N. Awano, K. Inubushi, E. Maeda, S. Nakamori, K. Nishino, A. Yamaguchi and H. Takagi: Effect of drug transporter genes on cysteine export and overproduction in Escherchia coli. Appl. Environ. Microbiol., 72, 4735-4742 (2006).
  11. Y. Kai, T. Kashiwagi, K. Ishikawa, M. K. Ziyatdinov, E. I. Redkina, M. Y. Kiriukhin, M. M. Gusyatiner, S. Kobayashi, H. Takagi and E. Suzuki: Engineering of Escherichia coli L-serine O-acetyltransferase on the basis of crystal structure: desensitization to feedback inhibition by L-cysteine. Protein Eng. Des. Sel., 19, 163-167 (2006).
  12. Y. Haitani, N. Awano, M. Yamazaki, M. Wada, S. Nakamori and H. Takagi: Functional analysis of L-serine O-acetyltransferase from Corynebacterium glutamicum. FEMS Microbiol. Lett., 255, 156-163 (2006).
  13. N. Awano, M. Wada, H. Mori, S. Nakamori and H. Takagi: Identification and Functional Analysis of Cysteine Desulfhydrases in Escherichia coli. Appl. Environ. Microbiol., 71, 4149-4152 (2005).
  14. S. Kobayashi, R. Masui, S. Yokoyama, S. Kuramitsu and H. Takagi: A novel metal-activated L-serine O-acetyltransferase from Thermus thermophilus HB8. J. Biochem., 136, 629-634 (2004).
  15. M. Wada, N. Awano, H. Yamazawa, H. Takagi and S. Nakamori: Purification and characterization of O-acetylserine sulfhydrylase of Corynebacterium glutamicum. Biosci. Biotech. Biochem., 68, 1581-1583 (2004).
  16. N. Awano, M. Wada, A. Kohdoh, T. Oikawa, H. Takagi and S. Nakamori: Effect of cysteine desulfhydrase gene disruption on L-cysteine overproduction in Escherichia coli. Appl. Microbiol. Biotechnol., 62, 239-243 (2003).
  17. H. Takagi, K. Yoshioka, N. Awano, S. Nakamori, and B.-I. Ono: Role of Saccharomyces cerevisiae serine O-acetyltransferase in cysteine biosynthesis. FEMS Microbiol. Lett., 218, 291-297 (2003).
  18. M. Wada, N. Awano, K. Haisa, H. Takagi and S. Nakamori: Purification, characterization and identification of cysteine desulfhydrase of Corynebacterium glutamicum, and its relationship to cysteine production. FEMS Microbiol. Lett., 217, 103-107 (2002).
  19. H. Takagi, N. Awano, S. Kobayashi, M. Noji, K. Saito and S. Nakamori: Overproduction of L-cysteine and L-cystine by expression of genes for feedback inhibition-insensitive serine acetyltransferase from Arabidopsis thaliana in Escherichia coli. FEMS Microbiol. Lett., 179, 453-459 (1999).
  20. S. Nakamori, S. Kobayashi, T. Nishimura and H. Takagi: Mechanism of L-methionine overproduction by Escherichia coli: the replacement of Ser54 by Asn in the MetJ protein causes the derepression of L-methionine biosynthetic enzymes. Appl. Microbiol. Biotechnol., 52, 179-185 (1999).
  21. H. Takagi, C. Kobayashi, S. Kobayashi and S. Nakamori: PCR random mutagenesis into Escherichia coli serine acetyltransferase: isolation of the mutant enzymes that cause overproduction of L-cysteine and L-cystine due to the desensitization to feedback inhibition. FEBS Lett., 452, 323-327 (1999).
  22. S. Nakamori, S. Kobayashi, C. Kobayashi and H. Takagi: Overproduction of L-cysteine and L-cystine by Escherichia coli strains with a genetically altered serine acetyltransferase. Appl. Environ. Microbiol., 64, 1607-1611 (1998).