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Molecular cloning of a phytase gene (phy M) from Pseudomonas syringae MOK1

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Abstract

A phytase gene (phy M) was cloned from Pseudomonas syringae MOK1 by two steps of degenerate PCR and inverse PCR. This gene consists of 1,287 nucleotides and encodes a polypeptide of 428 amino acids with a deduced molecular mass of 46,652 kDa. Based on its amino acid sequence, the Phy M shares the active site RHGXRXP and HD sequence motifs, typically characterized by histidine acid phosphatases familly. Each phy M gene fragment encoding mature Phy M with its own signal sequence (pEPSS) and without (pEPSM) was subcloned into the E. coli BL21 (DE3) expression vector, pET22b (+). The enzyme activity in crude extracts of clone pEPSM was 2.514 Umg−1 of protein, and about 10-fold higher than that of clone pEPSS.

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Acknowledgments

We are thankful for the graduate fellowship provided by the Ministry of Education through the Brain Korea 21 project.

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Authors and Affiliations

  1. Inositide Signaling Group, National Institute of Environmental Health Sciences, NIH, DHSS, Research Triangle Park, PO Box 12233, , NC, 27709, USA

    Jaiesoon Cho

  2. School of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, San 56-1 Sillim-dong, Gwanak-gu, Seoul, 151-742, Korea

    Changwhan Lee, Seungha Kang, Jaecheon Lee, Honggu Lee & Yunjaie Choi

  3. Choong Ang Biotech Co., Ltd., 833-6 Wonsi-Dong, Ansan-City, Kyunggi-Do, Korea

    Jinduck Bok

  4. National Institute of Mental Health, Bethesda, Maryland, 20892, USA

    Junghee Woo

  5. Jinju National University, 150 Chilamdong, Jinju, 660-758, Korea

    Yangsoo Moon

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  1. Jaiesoon Cho
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  2. Changwhan Lee
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  3. Seungha Kang
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  9. Yunjaie Choi
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Correspondence to Jaiesoon Cho.

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*These two authors have contributed equally to this work.

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Cho, J., Lee, C., Kang, S. et al. Molecular cloning of a phytase gene (phy M) from Pseudomonas syringae MOK1. Curr Microbiol 51, 11–15 (2005). https://doi.org/10.1007/s00284-005-4482-0

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  • DOI: https://doi.org/10.1007/s00284-005-4482-0

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