Skip to main content
SpringerLink
Log in

High production of laccase B from Trametes sp. in Pichia pastoris

  • Brief Communication
  • Published:
World Journal of Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

The coding sequence of a laccase isozyme from Trametes sp. AH28-2 was cloned in pPIC9K vector and heterologously overexpressed in the yeast Pichia pastoris strain GS115. In the minimal medium containing 0.3 mM CuSO4 and 0.6% alanine, the maximum yield of the recombinant laccase rLacB reached 32,000 U/l (1,012 U/mg), slightly higher than that of the native enzyme nLacB (∼30,000 U/l, 1,356 U/mg). The enzymatic properties of rLacB were different from those of nLacB as well. Regardless of the inferior thermal stability, rLacB had much better stability at both neutral and basic pH range compared to nLacB. In addition, the dye decolorization potential of rLacB was similar to that of nLacB.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

References

  • Abadulla E, Tzanov T, Costa S, Robra KH, Cavaco-Paulo A, Gübitz GM (2000) Decolorization and detoxification of textile dyes with a laccase from Trametes hirsuta. Appl Environ Microbiol 66:3357–3362

    Article  CAS  Google Scholar 

  • Bulter T, Alcalde M, Sieber V, Meinhold P, Schlachtbauer C, Arnold FH (2003) Functional expression of a fungal laccase in Saccharomyces cerevisiae by directed evolution. Appl Environ Microbiol 69:987–995

    Article  CAS  Google Scholar 

  • Chivukula M, Renganathan V (1995) Phenolic azo dye oxidation by laccase from Pyricularia oryzae. Appl Environ Microbiol 61:4374–4377

    CAS  Google Scholar 

  • Freire RS, Durán N, Kubota LT (2002) Development of a laccase-based flow injection electrochemical biosensor for phenolic compounds determination and its application for monitoring remediation of Kraft E1 paper mill effluent. Anal Chim Acta 463:229–238

    Article  CAS  Google Scholar 

  • Gianfreda L, Xu F, Bollag JM (1999) Laccases: a useful group of oxidoreductive enzymes. Biorem J 3:1–26

    Article  CAS  Google Scholar 

  • Han Y, Lei XG (1999) Role of glycosylation in the functional expression of an Aspergillus niger phytase gene (phyA) in Pichia pastoris. Arch Biochem Biophys 364:83–90

    Article  CAS  Google Scholar 

  • Hong F, Meinander NQ, Jönsson L (2002) Fermentation strategies for improved heterologous expression of laccase in Pichia pastoris. Biotechnol Bioeng 79:438–449

    Article  CAS  Google Scholar 

  • Hong YZ, Xiao YZ, Zhou HM, Fang W, Zhang M, Zhu J, Wang J, Wu LJ, Yu ZL (2006) Expression of a laccase cDNA from Trametes sp. AH28–2 in Pichia pastoris and mutagenesis of transformants by nitrogen ion implantation. FEMS Microbiol Lett 258:96–101

    Article  CAS  Google Scholar 

  • Hoshida H, Fujita T, Murata K, Kubo K, Akada R (2005) Copper-dependent production of a Pycnoporus coccineus extracellular laccase in Aspergillus oryzae and Saccharomyces cerevisiae. Biosci Biotechnol Biochem 69:1090–1097

    Article  CAS  Google Scholar 

  • Larsson S, Cassland P, Jönsson LJ (2001) Development of a Saccharomyces cerevisiae strain with enhanced resistance to phenolic fermentation inhibitors in lignocellulose hydrolysates by heterologous expression of laccase. Appl Environ Microbiol 67:1163–1170

    Article  CAS  Google Scholar 

  • Liu W, Chao Y, Liu S, Bao H, Qian S (2003) Molecular cloning and characterization of a laccase gene from the basidiomycete Fome lignosus and expression in Pichia pastoris. Appl Microbiol Biotechnol 63:174–181

    Article  CAS  Google Scholar 

  • Record E, Punt PJ, Chamkha M, Labat M, van Den Hondel CAMJJ, Asther M (2002) Expression of the Pycnoporus cinnabarinus laccase gene in Aspergillus niger and characterization of the recombinant enzyme. Eur J Biochem 269:602–609

    Article  CAS  Google Scholar 

  • Soden DM, Dobson ADW (2001) Differential regulation of laccase gene expression in Pleurotus sajor-caju. Microbiology 147:1755–1763

    CAS  Google Scholar 

  • Soden DM, O’Callaghan J, Dobson ADW (2002) Molecular cloning of a laccase isozyme gene from Pleurotus sajor-caju and expression in the heterologous Pichia pastoris host. Microbiology 148:4003–4014

    CAS  Google Scholar 

  • Thurston CF (1994) The structure and function of fungal laccases. Microbiology 140:19–26

    Article  CAS  Google Scholar 

  • Ullah MA, Bedford CT, Evans CS (2000) Reactions of pentachlorophenol with laccase from Coriolus versicolor. Appl Microbiol Biotechnol 53:230–234

    Article  CAS  Google Scholar 

  • Xiao YZ, Tu XM, Wang J, Zhang M, Cheng Q, Zeng WY, Shi YY (2003) Purification, molecular characterization and reactivity with aromatic compounds of a laccase from basidiomycete Trametes sp. strain AH28-2. Appl Microbiol Biotechnol 60:700–707

    CAS  Google Scholar 

  • Xiao YZ, Chen Q, Hang J, Shi YY, Wu J, Hong YZ, Wang YP (2004) Selective induction, purification and characterization of a laccase isozyme from the basidiomycete Trametes sp. AH28-2. Mycologia 96:26–35

    CAS  Google Scholar 

  • Xiao YZ, Zhou T, Hong YZ, Zhang M, Ge HH (2005) Dye decolorization and heterologous expression of laccase A from Trametes sp. AH28-2 In: Li SP (ed), Proceedings of the 8th internal conference on environmental microbes, Beijing, China: CI Press, pp 618–625

    Google Scholar 

  • Xiao YZ, Hong YZ, Li JF, Hang J, Tong PG, Fang W, Zhou CZ (2006) Cloning of novel laccase isozyme genes from Trametes sp. AH28-2 and analyzes of their differential expression. Appl Microbiol Biotechnol 71:493–501

    Article  CAS  Google Scholar 

  • Zhang H, Hong YZ, Xiao YZ, Yuan J, Tu XM, Zhang XQ (2006) Efficient production of laccases by Trametes sp. AH28-2 in co-cultivation with a Trichoderma strain. Appl Microbiol Biotechnol (in press). doi:10.1007/s00253-006-0430-6

Download references

Acknowledgements

The authors thank Dr. Jun Hang for the critical reading of the manuscript. This work was supported by grants from the National Natural Sciences Foundation of China (30670069, 30470056), the Science & Technology Foundation of Distinguished Young Scholars of Anhui Province (04043048, 05023057) and the Innovative Research Team of 211 Project in Anhui University (02203109).

Author information

Authors and Affiliations

  1. School of Life Science & Modern Experiment Technology Center, Anhui University, Hefei, 230039, P.R. China

    J. F. Li, Y. Z. Hong, Y. Z. Xiao, Y. H. Xu & W. Fang

Authors
  1. J. F. Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Y. Z. Hong
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Y. Z. Xiao
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Y. H. Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. W. Fang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Y. Z. Xiao.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, J.F., Hong, Y.Z., Xiao, Y.Z. et al. High production of laccase B from Trametes sp. in Pichia pastoris . World J Microbiol Biotechnol 23, 741–745 (2007). https://doi.org/10.1007/s11274-006-9286-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11274-006-9286-2

Keywords

Search

Navigation