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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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Literature Cited

  1. RM Berka MW Rey KM Brown T Byun AV Klotz (1998) ArticleTitleMolecular characterization and expression of a phytase gene from the thermophilic fungus Thermomyces lanuginosus Appl Environ Microbiol 64 4423–4427 Occurrence Handle9797301

    PubMed  Google Scholar 

  2. JS Cho CW Lee SH Kang JC Lee JD Bok YS Moon HG Lee SC Kim YJ Choi (2003) ArticleTitlePurification and characterization of a phytase from Pseudomonas syringae MOK1 Curr Microbiol 47 290–294 Occurrence Handle10.1007/s00284-002-3966-4 Occurrence Handle14629009

    Article  PubMed  Google Scholar 

  3. FH Common (1989) ArticleTitleBiological availability of phosphorus for pigs Nature 143 370–380

    Google Scholar 

  4. J Dassa C Marck PL Boquet (1990) ArticleTitleThe complete nucleotide sequence of the Escherichia coli app A reveals significant homology between pH 2.5 acid phosphatase and glucose-1-phosphatase J Bacteriol 172 5497–5500 Occurrence Handle2168385

    PubMed  Google Scholar 

  5. S Golovan G Wang J Zhang CW Forsberg (2000) ArticleTitleCharacterization and overproduction of the Escherichia coli appA encoded bifunctional enzyme that exhibits both phytase and acid phosphatase activities Can J Microbiol 46 59–71 Occurrence Handle10.1139/cjm-46-1-59 Occurrence Handle10696472

    Article  PubMed  Google Scholar 

  6. WV Hartingsveldt MJ Cora AM Cess JD Hodel (1993) ArticleTitleCloning, characterization and overexpression of the phytase-encoding gene (phyA) of Aspergillus niger Gene 127 87–94 Occurrence Handle10.1016/0378-1119(93)90620-I Occurrence Handle8387447

    Article  PubMed  Google Scholar 

  7. JK Heinonen RJ Lahti (1980) ArticleTitleA new and convenient colorimetric determination of inorganic orthophosphate and its application to the assay of inorganic pyrophosphatase Anal Biochem 113 313–317 Occurrence Handle10.1016/0003-2697(81)90082-8

    Article  Google Scholar 

  8. G Huang L Zhang RG Birch (2000) ArticleTitleRapid amplification and cloning of Tn5 flanking fragments by inverse PCR Lett Appl Microbiol 31 149–153 Occurrence Handle10.1046/j.1365-2672.2000.00781.x Occurrence Handle10972718

    Article  PubMed  Google Scholar 

  9. J Kerovuo M Lauraeus P Nurminen N Kalkkinen J Apajalahti (1998) ArticleTitleIsolation, characterization, molecular gene cloning, and sequencing of a novel phytase from Bacillus subtilis Appl Environ Microbiol 64 2079–2085 Occurrence Handle9603817

    PubMed  Google Scholar 

  10. OK Kim JK Lee HK Kim JH Yu TK Oh (1998) ArticleTitleCloning of the thermostable phytase gene(phy) from Bacillus sp. DS11 and its overexpression in Escherichia coli FEMS Microbiol Lett 162 185–191 Occurrence Handle10.1016/S0378-1097(98)00119-0 Occurrence Handle9595681

    Article  PubMed  Google Scholar 

  11. UK Laemmli (1970) ArticleTitleCleavage of structural proteins during the assembly of the head bacteriophage T4 Nature 227 680–685 Occurrence Handle5432063

    PubMed  Google Scholar 

  12. SF Lassen J Breinholt PR Ostergaard R Brugger A Bischoff M Wyss CC Fuglsang (2001) ArticleTitleExpression, gene cloning, and characterization of five novel phytases from four Basidiomycete Fungi: Peniophora lycii, Agrocybe pediades, a Ceriporia sp., and Trametes pubescens Appl Environ Microbiol 67 4701–4707 Occurrence Handle10.1128/AEM.67.10.4701-4707.2001 Occurrence Handle11571175

    Article  PubMed  Google Scholar 

  13. T Maniatis EF Fritsch J Sambrook (1989) Molecular Cloning Laboratory Manual EditionNumber2 Cold Spring Harbor Laboratory Press Plainview, NY

    Google Scholar 

  14. DB Mitchell K Vogel BJ Weimann L Pasamontes APGM Loon Particlevan (1997) ArticleTitleThe phytase subfamily of histidine acid phosphatases:isolation of genes for two novel phytases from the fungi Aspergillus terreus and Myceliophthora thermophila Microbiol 143 245–252 Occurrence Handle10.1016/0923-2508(92)90016-H

    Article  Google Scholar 

  15. EJ Mullaney DM Gibson AHJ Ullah (1991) ArticleTitlePositive identification of a lambda gt11 clone containing a region of fungal phytase gene by immunoprobe and sequence verification Appl Microbiol Biotechnol 35 611–614 Occurrence Handle10.1007/BF00169625 Occurrence Handle1369340

    Article  PubMed  Google Scholar 

  16. L Pasamontes M Haiker M Wyss M Henriquez-Huecas DB Mitchell APGM Loon Particlevan (1997) ArticleTitleCloning of the phytases from Emericella nidulans and the thermophilic fungus Talaromyces thermophilus Biochim Biophys Acta 1353 217–223 Occurrence Handle9349716

    PubMed  Google Scholar 

  17. L Pasamontes M Haiker M Wyss M Tessier APGM Loon Particlevan (1997) ArticleTitleGene cloning, purification and characterization of a heat-stable phytase from the fungus Aspergillus fumigatus Appl Environ Microbiol 63 1696–1700 Occurrence Handle9143104

    PubMed  Google Scholar 

  18. J Pen TC Verwoerd A Hoekema (1993) ArticleTitlePhytase-containing transgenic seeds as novel feed additive for improved phosphorus utilization Biotechnology 11 811–814 Occurrence Handle10.1038/nbt0793-811

    Article  Google Scholar 

  19. CS Piddington CS Houston M Paloheimo M Cantrell A Miettinen-oinonen A Nevalainen J Rambosek (1993) ArticleTitleThe cloning and sequencing of the genes encoding phytase (phy) and pH 2.5-optimum acid phosphatase (aph) from Aspergillus niger var awamori. Gene 133 166–171

    Google Scholar 

  20. NR Reddy SK Sathe DK Salunkhe (1989) ArticleTitlePhytases in legumes and cereals Adv Food Res 28 1–92

    Google Scholar 

  21. DP Roberts TP Denny MA Schell (1988) ArticleTitleCloning of the egl gene of Pseudomonas solanacearum and analysis of its role in phytopathogenicity J. Bacteriol 170 1445–1451 Occurrence Handle2832363

    PubMed  Google Scholar 

  22. E Rodriquez Y Han XG Lei (1999) ArticleTitleCloning, sequencing and expression of an Escherichia coli acid phosphatase/phytase gene (appA2) isolated from pig colon BBRC 257 117–123 Occurrence Handle10092520

    PubMed  Google Scholar 

  23. A Sajidan A Farouk R Greiner P Jungblut EC Muller R Borris (2004) ArticleTitleMolecular and physiological characterization of a 3-phytase from soil bacterium Klebsiella sp. ASR1 Appl Microbiol Biotechnol 65 110–118 Occurrence Handle10.1007/s00253-003-1530-1 Occurrence Handle14727093

    Article  PubMed  Google Scholar 

  24. PC Simon HAJ Versteegh (1990) ArticleTitleImprovement of phosphorus availability by microbial phytase in broilers and pigs Br J Nutr 64 525–540 Occurrence Handle2171634

    PubMed  Google Scholar 

  25. AJ Tye FKY Siu TYC Leung BL Lim (2002) ArticleTitleMolecular cloning and the biochemical characterization of two novel phytases from B. subtilis 168 and B. licheniformis Appl Microbiol Biotechnol 59 190–197 Occurrence Handle10.1007/s00253-002-1033-5 Occurrence Handle12111145

    Article  PubMed  Google Scholar 

  26. NV Zinin AV Serkina MS Gelfand AB Shevelev SP Sineoky (2004) ArticleTitleGene cloning, expression and characterization of novel phytase from Obesumbacterium proteus FEMS Microbiol Lett 236 283–290 Occurrence Handle10.1016/j.femsle.2004.05.051 Occurrence Handle15251209

    Article  PubMed  Google Scholar 

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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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  5. Honggu Lee
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  8. Yangsoo Moon
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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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