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树皮刮取真菌:应用于非纺织染料生物修复的漆酶潜在来源。

Tree bark scrape fungus: A potential source of laccase for application in bioremediation of non-textile dyes.

机构信息

Department of Microbiology, PSGVP Mandal's Arts, Science and Commerce College, Shahada, India.

Department of Biotechnology, SSVP Sansth's Late Karmveer Dr. P. R. Ghogrey Science College, Dhule, India.

出版信息

PLoS One. 2020 Jun 4;15(6):e0229968. doi: 10.1371/journal.pone.0229968. eCollection 2020.

Abstract

Although laccase has been recognized as a wonder molecule and green enzyme, the use of low yielding fungal strains, poor production, purification, and low enzyme kinetics have hampered its large-scale application. Thus,this study aims to select high yielding fungal strains and optimize the production, purification, and kinetics of laccase of Aspergillus sp. HB_RZ4. The results obtained indicated that Aspergillus sp. HB_RZ4 produced a significantly large amount of laccase under meso-acidophilic shaking conditions in a medium containing glucose and yeast extract. A 25 μM CuSO4 was observed to enhance the enzyme yield. The enzyme was best purified on a Sephadex G-100 column. The purified enzyme resembled laccase of A. flavus. The kinetics of the purified enzyme revealed high substrate specificity and good velocity of reaction,using ABTS as a substrate. The enzyme was observed to be stable over various pH values and temperatures. The peptide structure of the purified enzyme was found to resemble laccase of A. kawachii IFO 4308. The fungus was observed to decolorize various dyes independent of the requirement of a laccase mediator system.Aspergillus sp. HB_RZ4 was observed to be a potent natural producer of laccase, and it decolorized the dyes even in the absence of a laccase mediator system. Thus, it can be used for bioremediation of effluent that contains non-textile dyes.

摘要

尽管漆酶已被视为神奇分子和绿色酶,但由于产量低的真菌菌株、生产、纯化和低酶动力学的限制,其大规模应用受到了阻碍。因此,本研究旨在选择高产真菌菌株,并优化 Aspergillus sp. HB_RZ4 漆酶的生产、纯化和动力学。研究结果表明,Aspergillus sp. HB_RZ4 在含有葡萄糖和酵母提取物的中酸性摇瓶条件下,能产生大量漆酶。实验发现 25μM 的 CuSO4 能提高酶的产量。酶的最佳纯化方法是使用 Sephadex G-100 柱。纯化后的酶与 A. flavus 的漆酶相似。用 ABTS 作为底物时,该酶的动力学表现出高底物特异性和良好的反应速度。该酶在各种 pH 值和温度下均表现出稳定性。纯化后的酶的肽结构与 A. kawachii IFO 4308 的漆酶相似。该真菌能够独立于漆酶介体系统对各种染料进行脱色。Aspergillus sp. HB_RZ4 是一种潜在的天然漆酶产生菌,即使在没有漆酶介体系统的情况下,它也能对染料进行脱色。因此,它可以用于含有非纺织染料的废水的生物修复。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0dee/7272029/9e0cf29d33a1/pone.0229968.g001.jpg

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