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Optimization of the expression of a laccase gene from Trametes versicolor in Pichia methanolica

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Abstract

A cDNA encoding for laccase (Lcc1) was isolated from the ligninolytic fungus Trametes versicolor by reverse transcriptase polymerase chain reaction. The Lcc1 gene was subcloned into the Pichia methanolica expression vector pMETαA and transformed into the P. methanolica strains PMAD11 and PMAD16. The extracellular laccase activity of the PMAD11 recombinants was found to be 1.3-fold higher than that of the PMAD16 recombinants. The identity of the recombinant protein was further confirmed by immunodetection using the Western blot analysis. As expected, the molecular mass of the mature laccase was 64.0 kDa, similar to that of the native form. The effects of copper concentration, cultivation temperature, pH and methanol concentration in the BMMY on laccase expression were investigated. The laccase activity in the PMAD11 recombinant was up to 12.6 U ml−1 by optimization.

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Authors and Affiliations

  1. Key Laboratory of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300222, PR China

    Mei Guo, Fuping Lu, Lianxiang Du, Jun Pu & Dongqing Bai

  2. Department of Food Science, Tianjin Agriculture College, Tianjin, 300384, PR China

    Mei Guo

  3. Key Laboratory of Ecology and Aquaculture, Department of Fishery Science, Tianjin Agriculture College, Tianjin, 300384, PR China

    Dongqing Bai

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  1. Mei Guo
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  2. Fuping Lu
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  3. Lianxiang Du
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  4. Jun Pu
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  5. Dongqing Bai
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Correspondence to Fuping Lu.

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Guo, M., Lu, F., Du, L. et al. Optimization of the expression of a laccase gene from Trametes versicolor in Pichia methanolica . Appl Microbiol Biotechnol 71, 848–852 (2006). https://doi.org/10.1007/s00253-005-0210-8

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  • DOI: https://doi.org/10.1007/s00253-005-0210-8

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