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Heterologous expression and characterisation of a laccase from Colletotrichum lagenarium and decolourisation of different synthetic dyes

文献类型： 外文期刊

作者： Wang, Bo 1 ; Yan, Ying 2 ; Tian, Yongsheng 1 ; Zhao, Wei 1 ; Li, Zhengjun 1 ; Gao, Jianjie 1 ; Peng, Rihe 1 ; Yao, Quanho 1 ;

作者机构： 1.Shanghai Acad Agr Sci, Biotechnol Res Inst, Shanghai 201106, Peoples R China

2.Shanghai Acad Agr Sci, Crop Breeding & Cultivat Res Inst, Shanghai 201403, Peoples R China

关键词： Laccase;Colletotrichum lagenarium;Pichia pastoris;Characterisation;Synthetic dyes

期刊名称：WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY （ 影响因子：3.312； 五年影响因子：3.58 ）

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收录情况： SCI

摘要： Laccases have received considerable attention in recent decades because of their ability to oxidise a large spectrum of phenolic and non-phenolic organic substrates and highly recalcitrant environmental pollutants. In this research, a laccase gene from Colletotrichum lagenarium was chemically synthesised using yeast bias codons and expressed in Pichia pastoris. The molecular mass of the recombinant laccase was estimated to be 64.6 kDa by SDS-PAGE, and the enzyme exhibited maximum activity at pH 3.6-4.0 but more stability in buffer with higher pH (>pH 3.6). The optimal reaction temperature of the enzyme was 40 degrees C, beyond which stability significantly decreased. By using 2,20'- azino-bis-(3-ethylbenzothiazoline)-6-sulphonate (ABTS) as a substrate, K-m and V-max values of 0.34 mM and 7.11 mM min(-1) mg(-1), respectively, were obtained. Using ABTS as a mediator, the laccase could oxidise hydroquinone to p-benzoquinone and decolourise the synthetic dyes malachite green, crystal violet and orange G. These results indicated that the laccase could be used to treat industrial effluents containing artificial dyes.

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