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Cloning of novel bacterial xylanases from lignocellulose-enriched compost metagenomic libraries

机译:从富含木质纤维素的堆肥宏基因组学文库中克隆新型细菌木聚糖酶

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摘要

Xylanases are in important class of industrial enzymes that are essential for the complete hydrolysis of lignocellulosic biomass into fermentable sugars. In the present study, we report the cloning of novel xylanases with interesting properties from compost metagenomics libraries. Controlled composting of lignocellulosic materials was used to enrich the microbial population in lignocellulolytic organisms. DNA extracted from the compost samples was used to construct metagenomics libraries, which were screened for xylanase activity. In total, 40 clones exhibiting xylanase activity were identified and the thermostability of the discovered xylanases was assayed directly from the library clones. Five genes, including one belonging to the more rare family GH8, were selected for subcloning and the enzymes were expressed in recombinant form in E. coli. Preliminary characterization of the metagenome-derived xylanases revealed interesting properties of the novel enzymes, such as high thermostability and specific activity, and differences in hydrolysis profiles. One enzyme was found to perform better than a standard Trichoderma reesei xylanase in the hydrolysis of lignocellulose at elevated temperatures.
机译:木聚糖酶是工业酶的重要类别,对于木质纤维素生物质完全水解为可发酵糖至关重要。在本研究中,我们报道了堆肥宏基因组学库中具有有趣特性的新型木聚糖酶的克隆。木质纤维素材料的受控堆肥用于富集木质纤维素分解生物中的微生物种群。从堆肥样品中提取的DNA用于构建宏基因组学文库,并筛选木聚糖酶活性。总共鉴定出40个表现出木聚糖酶活性的克隆,并直接从文库克隆中分析发现的木聚糖酶的热稳定性。选择了五个基因,包括一个属于较罕见的GH8家族的基因,进行亚克隆,并在大肠杆菌中以重组形式表达了这些酶。源自元基因组的木聚糖酶的初步表征揭示了新型酶的有趣特性,例如高热稳定性和比活性,以及​​水解谱的差异。发现一种酶在升高的温度下水解木质纤维素时比标准的里氏木霉木聚糖酶表现更好。

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