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Optimisation of the Production and Bleaching Process for a New Laccase from Madurella mycetomatis , Expressed in Pichia pastoris : from Secretion to Yielding Prominent
Molecular Biotechnology ( IF 2.4 ) Pub Date : 2020-10-15 , DOI: 10.1007/s12033-020-00281-9
Ahmet Tülek 1 , Ersin Karataş 1 , Mehmet Mervan Çakar 1 , Derya Aydın 2 , Özlem Yılmazcan 2 , Barış Binay 3
Affiliation  

Laccases are polyphenol oxidoreductases used in a number of industrial applications. Due to the increasing demand for these “green catalysis” enzymes, the identification and biochemical characterisation of their novel properties is essential. In our study, cloned Madurella mycetomatis laccase (mmlac) genes were heterologously expressed in the methylotrophic yeast host Pichia pastoris. The high yield of the active recombinant protein in P. pastoris demonstrates the efficiency of a reliably constructed plasmid to express the laccase gene. The optimal biochemical conditions for the successfully expressed MmLac enzyme were identified. Detailed structural properties of the recombinant laccase were determined, and its utility in decolourisation and textile bleaching applications was examined. MmLac demonstrates good activity in an acidic pH range (4.0–6.0); is stable in the presence of cationic metals, organic solvents and under high temperatures (50–60 °C); and is stable for long-term storage at − 20 °C and − 80 °C for up to eight weeks. The structural analysis revealed that the catalytic residues are partially similar to other laccases. MmLac resulted in an increase in whiteness, whilst demonstrating high efficiency and stability and requiring the input of fewer chemicals. The performance of this enzyme makes it worthy of investigation for use in textile biotechnology applications, as well as within environmental and food technologies.



中文翻译:

巴氏毕赤酵母新漆酶的生产和漂白工艺优化:从分泌到产量突出

漆酶是在许多工业应用中使用的多酚氧化还原酶。由于对这些“绿色催化”酶的需求不断增加,对其新特性的鉴定和生化表征至关重要。在我们的研究中,克隆的Madurella mycetomatis漆酶 ( mmlac ) 基因在甲基营养酵母宿主毕赤酵母中异源表达。巴斯德毕赤酵母中活性重组蛋白的高产量证明了可靠构建的质粒表达漆酶基因的效率。成功表达Mm的最佳生化条件鉴定了紫胶酶。确定了重组漆酶的详细结构特性,并检查了其在脱色和纺织品漂白应用中的效用。Mm Lac 在酸性 pH 值范围 (4.0–6.0) 中表现出良好的活性;在阳离子金属、有机溶剂和高温(50-60°C)下稳定;在 − 20 °C 和 − 80 °C 下可长期储存长达八周。结构分析显示催化残基与其他漆酶部分相似。毫米紫胶导致白度增加,同时表现出高效率和稳定性,并且需要更少的化学品输入。这种酶的性能使其在纺织生物技术应用以及环境和食品技术中的应用值得研究。

更新日期:2020-10-15
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