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CysK from Lactobacillus casei encodes a protein with O-acetylserine sulfhydrylase and cysteine desulfurization activity

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Abstract

A gene encoding an O-acetyl-l-serine sulfhydrylase (cysK) was cloned from Lactobacillus casei FAM18110 and expressed in Escherichia coli. The purified recombinant enzyme synthesized cysteine from sulfide and O-acetyl-l-serine at pH 5.5 and pH 7.4. At pH 7.4, the apparent K M for O-acetyl-l-serine (OAS) and sulfide were 0.6 and 6.7 mM, respectively. Furthermore, the enzyme showed cysteine desulfurization activity in the presence of dithiothreitol at pH 7.5, but not at pH 5.5. The apparent K M for l-cysteine was 0.7 mM. The synthesis of cystathionine from homocysteine and serine or OAS was not observed. When expressed in a cysMK mutant of Escherichia coli, the cloned gene complemented the cysteine auxotrophy of the mutant. These findings suggested that the gene product is mainly involved in cysteine biosynthesis in L. casei. Quantitative real-time PCR and a mass spectrometric assay based on selected reaction monitoring demonstrated that L. casei FAM18110 is constitutively overexpressing cysK.

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Acknowledgment

We thank Dr. Naoko Yoshimoto from the Chiba University in Japan for providing the E. coli NK3 strain. Furthermore, we thank T. Bavan and M. Haueter for providing technical assistance.

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Authors and Affiliations

  1. Agroscope Liebefeld-Posieux Research Station ALP, Bern, Switzerland

    Biljana Bogicevic, Hélène Berthoud, Reto Portmann & Stefan Irmler

  2. ETH Zurich, Institute of Food, Nutrition and Health (IFNH), Laboratory of Food Biotechnology, Zürich, Switzerland

    Biljana Bogicevic & Leo Meile

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  1. Biljana Bogicevic
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  2. Hélène Berthoud
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  4. Leo Meile
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Correspondence to Stefan Irmler.

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Bogicevic, B., Berthoud, H., Portmann, R. et al. CysK from Lactobacillus casei encodes a protein with O-acetylserine sulfhydrylase and cysteine desulfurization activity. Appl Microbiol Biotechnol 94, 1209–1220 (2012). https://doi.org/10.1007/s00253-011-3677-5

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  • DOI: https://doi.org/10.1007/s00253-011-3677-5

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