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Cloning of novel laccase isozyme genes from Trametes sp. AH28-2 and analyses of their differential expression

  • Applied Genetics and Molecular Biotechnology
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Abstract

Three novel laccase isozyme genes, lacA, lacB, and lacC, have been identified from basidiomycete Trametes sp. AH28-2. These genes display a high similarity with other basidiomycete laccases at the amino acid level. An inferred TATA box and several putative CAAT, MRE, XRE, and CreA consensus sequences were identified in the lacA, lacB, and lacC promoter regions. Different from the TATA boxes of lacA and lacB at about −100, the TATA box of lacC is located at −172. For all the isozymes, copper ion is essential for laccase synthesis in Trametes sp. AH28-2. More interestingly, different aromatic compounds can selectively induce the production of distinct laccase isozymes, with o-toluidine inducing the expression of laccase A (LacA) while 3,5-dihydroxytoluene mainly stimulating the production of laccase B (LacB). Quantitative reverse transcriptase–polymerase chain reaction showed that the accumulation of laccase messenger RNA transcripts is accompanied by the increase of corresponding enzyme activity in cultures. The glucose-repression effect on laccase expression in Trametes sp. AH28-2 was also observed. Furthermore, lower Cu2+ concentration (lower than 0.5 mM) can induce LacA and a novel laccase (LacC), and the latter will disappear when Cu2+ concentration is increased up to 1−2 mM. Upon induction by 3,5-dihydroxytoluene, the ratio of LacA to LacB decreased in the later phase of induction.

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Acknowledgements

This work was supported by grants from the National Natural Sciences Foundation of China (30370045, 30470056), Key Programs for Science and Technology of Chinese Ministry of Education (204064), Science & Technology Foundation of Distinguished Young Scholars of Anhui Province (04043048), and Innovative Research Team of 211 Project in Anhui University.

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Authors and Affiliations

  1. School of Life Science & Modern Experiment Center, Anhui University, 230039, Hefei, People's Republic of China

    Y. Z. Xiao, Y. Z. Hong, J. F. Li, P. G. Tong & W. Fang

  2. Creatv MicroTech, Inc., 11609 Lake Potomac Drive, Potomac, MD, 20854, USA

    J. Hang

  3. Laboratory of Structure Biology, School of Life Sciences, University of Science and Technology of China, 230026, Hefei, People's Republic of China

    C. Z. Zhou

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  1. Y. Z. Xiao
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  2. Y. Z. Hong
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  3. J. F. Li
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Correspondence to Y. Z. Xiao.

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Xiao, Y.Z., Hong, Y.Z., Li, J.F. et al. Cloning of novel laccase isozyme genes from Trametes sp. AH28-2 and analyses of their differential expression. Appl Microbiol Biotechnol 71, 493–501 (2006). https://doi.org/10.1007/s00253-005-0188-2

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  • DOI: https://doi.org/10.1007/s00253-005-0188-2

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