Characterization and Degradation of Triphenylmethane Dyes and Their Leuco-Derivatives by Heterologously Expressed Laccase From Coprinus cinerea

文献类型： 外文期刊

第一作者： Qian, Cen

作者： Qian, Cen;Wang, Bo;Peng, Rihe;Yao, Quanhong;Pei, Zuodi

作者机构：

关键词： Coprinus cinerea; laccase; leuco-derivatives; Pichia pastoris; triphenylmethane dyes

期刊名称：CELL BIOCHEMISTRY AND FUNCTION （ 影响因子：2.7； 五年影响因子：3.0 ）

ISSN： 0263-6484

年卷期： 2024 年 42 卷 7 期

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收录情况： SCI

摘要： Laccase is a copper-containing polyphenol oxidase that can oxidize phenolic and non-phenolic organic substrates. In the past decades, laccases had received considerable attention because of the ability to degrade various organic substances. Based on the codon preference of the Pichia pastoris expression system, this study optimized the gene structure of the laccase gene Lcc1 from Coprius cinerea through synthetic biology methods. A new gene Lcc1I was synthesized and heterologously expressed in P. pastoris. After 3 days of cultivation in a shake flask at 30 degrees C, the transformants produced at a yield of 890 mg L(-1)protein. The highest production level of the recombinant laccase was 2760 U L-1. The molecular mass of the recombinant laccase was estimated at 60 kDa. The enzyme showed highest activity at pH 3.4 and 45 degrees C. It possessed better stability at higher pH and lower temperature condition. Using 2,2'-azino-bis-(3-ethylbenzothiazoline)-6-sulphonate (ABTS) as the substrate, the K-m and V-max values were 0.136 mM and 9778 mu M min(-1) mg(-1), respectively. The recombinant laccase could directly oxidize some triphenylmethane dyes like leuco-crystal violet (LCV) and leuco-malachite green (LMG). With the help of ABTS mediator, it could oxidize and degrade 77.7% crystal violet (CV) and 79.2% malachite green (MG) within 1 h. Our results indicate that optimization of the laccase gene achieves good expression results in the host system. The dye degradation model constructed in this study may also be applied to the degradation of other organic pollutants and toxic substances, providing new solutions for environmental remediation against the increasingly severe environmental pollution.

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