Molecular clustering of Pycnoporus strains from various geographic origins and isolation of monokaryotic strains for laccase hyperproduction

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• 作者：Anne Lomascolo ; Jean-Luc Cayol ; Marjolaine Roche ; Lin Guo ; Jean-Luc Robert ; Eric Record ; Laurence Lesage-Meessen ; Bernard Ollivier ; Jean-Claude Sigoillot ; Marcel Asther
• 刊名：Mycological Research
• 出版年：2002
• 出版时间：October 2002
• 年：2002
• 卷：106
• 期：10
• 页码：1193-1203
• 全文大小：209 K

文摘

The production of laccase, an enzyme of industrial interest, was screened among species of the genus Pycnoporus, in particular P. sanguineus. Strains were isolated from various tropical Chinese environments and phylogenetically compared to ones deposited in international collections. Molecular clustering, based on ribosomal ITS1–5.8S-ITS2 genomic sequence analysis, showed that the Chinese strains of P. sanguineus formed an homogeneous phylogenetic group distinguished by its laccase-overproducing character. The dikaryotic strain P. sanguineus G05 was selected for its ability to produce up to 40000 U 1⁻¹ laccase in the presence of 2,5-xylidine, Tween 80 and maize bran. Since fruit bodies of P. sanguineus could be formed in the laboratory, monokaryotic laccase-hyperproducing strains were isolated using classic genetical methods. Among these isolates, strain G05.10 synthesized up to 71 000 U 1⁻¹ laccase, with a productivity of 5069 U 1⁻¹ d⁻¹. The laccase was purified and identified as a 70 kDa protein with an acidic pI, and was very stable at high temperatures.