拟南芥 MUT3379:高产漆酶的新型细胞工厂

Luca Mellere, Martina Bellasio, Francesca Berini, Flavia Marinelli, Jean Armengaud, F. Beltrametti
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摘要

Coriolopsis trogii 是一种基生真菌,利用大量木质素修饰酶来定殖和分解枯木。它的细胞外酶库包括长效酶(即多酚氧化酶),这种酶具有氧化各种天然和合成化合物的能力,因此在不同的工业应用中具有重要意义。在这项工作中,我们探索并改进了 C. trogii MUT3379 中漆化酶的生产。发酵过程从最初的约 10,000 U L-1 的产量逐渐优化,最终达到约 200,000 U L-1 的产量。分泌物组的 SDS-PAGE 分析显示,有一种 60 kDa 左右的主要蛋白质具有漆酶活性,通过串联质谱确定其主序列后,将其命名为 Lac3379-1。对 Lac3379-1 的表征表明,它在表面活性剂和溶剂存在下具有显著的酶稳定性,并在多种底物上具有多样化的活性,使其成为一种可用于多种生物技术应用的有趣工具。高产和稳健的生产过程表明,C. trogii MUT3379 是一种很有前途的漆酶细胞工厂,为木质素改性酶的工业应用提供了新的前景。
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Coriolopsis trogii MUT3379: A Novel Cell Factory for High-Yield Laccase Production
Coriolopsis trogii is a basidiomycete fungus which utilizes a large array of lignin-modifying enzymes to colonize and decompose dead wood. Its extracellular enzymatic arsenal includes laccases, i.e., polyphenol oxidases of relevant interest for different industrial applications thanks to their ability to oxidize a diverse range of natural and synthetic compounds. In this work, the production of laccases in C. trogii MUT3379 was explored and improved. From an initial production of ca. 10,000 U L−1, the fermentation process was gradually optimized, reaching a final yield of ca. 200,000 U L−1. An SDS-PAGE analysis of the secretome highlighted the presence of a main protein of ca. 60 kDa showing laccase activity, which was designated as Lac3379-1 once its primary sequence was established by tandem mass spectrometry. The characterization of Lac3379-1 revealed a remarkable enzymatic stability in the presence of surfactants and solvents and a diversified activity on a broad range of substrates, positioning it as an interesting tool for diverse biotechnological applications. The high-yield and robust production process indicates C. trogii MUT3379 as a promising cell factory for laccases, offering new perspectives for industrial applications of lignin-modifying enzymes.
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