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Cloning, characterization and expression of two new polyphenol oxidase cDNAs from Agaricus bisporus

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Abstract

Two new polyphenol oxidase (PPO) cDNAs (PPO3 and PPO4 cDNAs, accession numbers GQ354801 and GQ354802, respectively) were obtained by RACE-PCR from Agaricus bisporus. PPO3 cDNA was 1844 bp in length with an open reading frame of 1731 bp, while PPO4 cDNA was 2042 bp with an open reading frame of 1836 bp. PPO3 and PPO4 cDNAs, with 52% identity at the nucleic acid level, encoded a 576-amino acid protein of 66.3 kDa and 611-amino acid protein of 68.3 kDa, respectively. Mature forms of PPO3 and PPO4 were characterized after removing the specific C-terminal region and expressed in Escherichia coli BL21 (DE3) RIPL using pGEX-4T-1 vector. The expressed proteins were probed by the anti-A. bisporus PPO antibody but without PPO activity. This indicated that the recombinant mature PPO3 and mature PPO4 could not form an active center in prokaryotic expression system.

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References

  • Coseteng MY, Lee CY (1987) Changes in apple polyphenol oxidase and polyphenol concentrations in relation to degree of browning. J Food Sci 52:985–989

    Article  CAS  Google Scholar 

  • Fan Y, Flurkey WH (2004) Purification and characterization of tyrosinase from gill tissue of Portabella mushrooms. Phytochemistry 65:671–678

    Article  CAS  PubMed  Google Scholar 

  • Fraignier MP, Marques L, Fleuriet A, Macheix JJ (1995) Biochemical and immunochemical characteristics of polyphenol oxidases from different fruits of Prunus. J Agric Food Chem 43:2375–2380

    Article  CAS  Google Scholar 

  • Fujita Y, Uraga Y, Ichisima E (1995) Molecular cloning and nucleotide sequence of the protyrosinase gene, melO, from Aspergillus oryzae and expression of the gene in yeast cells. Biochim Biophys Acta Gene Struct Expr 1261:151–154

    Google Scholar 

  • Garcia-Borron JC, Solano F (2002) Molecular anatomy of tyrosinase and its related proteins: beyond the histidine-bound metal catalytic center. Pigment Cell Res 15:162–173

    Article  CAS  PubMed  Google Scholar 

  • Gawlik-Dziki U, Zlotek U, Swieca M (2008) Characterization of polyphenol oxidase from butter lettuce (Lactuca sativa var. capitata L.). Food Chem 107:129–135

    Article  CAS  Google Scholar 

  • Gerdemann C, Eicken C, Krebs B (2002) The crystal structure of catechol oxidase: new insight into the function of type-3 copper proteins. Acc Chem Res 35:183–191

    Article  CAS  PubMed  Google Scholar 

  • Halaouli S, Record E, Casalot L, Hamdi M, Sigoillot JC, Asther M, Lomascolo A (2006) Cloning and characterization of a tyrosinase gene from the white-rot fungus Pycnoporus sanguineus, and overproduction of the recombinant protein in Aspergillus niger. Appl Microbiol Biotechnol 70:580–589

    Article  CAS  PubMed  Google Scholar 

  • Kawamura-Konishi Y, Tsuji M, Hatana S, Asanuma M, Kakuta D, Kawano T, Mukouyama EB, Goto H, Suzuki H (2007) Purification, characterization, and molecular cloning of tyrosinase from Pholiota nameko. Biosci Biotechnol Biochem 71:1752–1760

    Article  CAS  PubMed  Google Scholar 

  • Kupper U, Niedermann DM, Travaglini G, Lerch K (1989) Isolation and characterization of the tyrosinase gene from Neurospora crassa. J Biol Chem 264:17250–17258

    CAS  PubMed  Google Scholar 

  • Lantto R, Plathin P, Niemist M, Buchert J, Autio K (2006) Effects of transglutaminase, tyrosinase and freeze-dried apple pomace powder on gel forming and structure of pork meat. LWT Food Sci Technol 39:1117–1124

    Article  CAS  Google Scholar 

  • Madani W, Kermasha S, Goetghebeur M, Tse M (1997) Partial purification and characterization of a polyphenol esterase from Aspergillus niger. Process Biochem 32:61–69

    Article  CAS  Google Scholar 

  • Marusek CM, Trobaugh NM, Flurkey WH, Inlow JK (2006) Comparative analysis of polyphenol oxidase from plant and fungal species. J Inorg Biochem 100:108–123

    Article  CAS  PubMed  Google Scholar 

  • Mayer AM, Harel E (1979) Polyphenol oxidase in plants. Phytochemistry 18:193–215

    Article  CAS  Google Scholar 

  • Takasaki S, Kawakishi S, Murata M, Homma S (2001) Polymerisation of gliadin mediated by mushroom tyrosinase. LWT Food Sci Technol 34:507–512

    CAS  Google Scholar 

  • Thalmann CR, Lötzbeyer T (2002) Enzymatic cross-linking of proteins with tyrosinase. Eur Food Res Technol 214:276–281

    Article  CAS  Google Scholar 

  • Wichers HJ, Gerritsen YAM, Chapelon CGJ (1996) Tyrosinase isoforms from the fruitbodies of Agaricus bisporus. Phytochemistry 43:333–337

    Article  CAS  Google Scholar 

  • Wichers HJ, Recourt K, Hendriks M, Ebbelaar CE, Biancone G, Hoeberichts FA, Mooibroek H, Soler-Rivas C (2003) Cloning, expression and characterisation of two tyrosinase cDNAs from Agaricus bisporus. Appl Microbiol Biotechnol 61:336–341

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by the National High Technology Research and Development Program of China (863 Program) (No.2006AA10Z324) and Goal-oriented Project of State Key Laboratory of Food Science and Technology, Nanchang University (No. SKLF-MB-200807).

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Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047, China

    Jinju Wu, Hongbing Chen, Jinyan Gao, Xiao Liu, Wei Cheng & Xiaojuan Ma

  2. Sino-German Joint Research Institute, Nanchang University, Nanchang, 330047, China

    Hongbing Chen

  3. Department of Food Science, Nanchang University, Nanchang, 330047, China

    Jinyan Gao

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  1. Jinju Wu
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  2. Hongbing Chen
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  3. Jinyan Gao
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  4. Xiao Liu
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  5. Wei Cheng
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Correspondence to Jinyan Gao.

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Wu, J., Chen, H., Gao, J. et al. Cloning, characterization and expression of two new polyphenol oxidase cDNAs from Agaricus bisporus . Biotechnol Lett 32, 1439–1447 (2010). https://doi.org/10.1007/s10529-010-0329-2

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  • DOI: https://doi.org/10.1007/s10529-010-0329-2

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