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Glycoside hydrolase activities of thermophilic bacterial consortia adapted to switchgrass

机译:适应柳枝thermo的嗜热细菌群落糖苷水解酶活性

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摘要

Industrial-scale biofuel production requires robust enzymatic cocktails to produce fermentable sugars from lignocellulosic biomass. Thermophilic bacterial consortia are a potential source of cellulases and hemicellulases adapted to harsher reaction conditions than commercial fungal enzymes. Compost-derived microbial consortia were adapted to switchgrass at 60°C to develop thermophilic biomass-degrading consortia for detailed studies. Microbial community analysis using small-subunit rRNA gene amplicon pyrosequencing and short-read metagenomic sequencing demonstrated that thermophilic adaptation to switchgrass resulted in low-diversity bacterial consortia with a high abundance of bacteria related to thermophilic paenibacilli, Rhodothermus marinus, and Thermus thermophilus. At lower abundance, thermophilic Chloroflexi and an uncultivated lineage of the Gemmatimonadetes phylum were observed. Supernatants isolated from these consortia had high levels of xylanase and endoglucanase activities. Compared to commercial enzyme preparations, the endoglucanase enzymes had a higher thermotolerance and were more stable in the presence of 1-ethyl-3-methylimidazolium acetate ([C2mim][OAc]), an ionic liquid used for biomass pretreatment. The supernatants were used to saccharify [C2mim][OAc]-pretreated switchgrass at elevated temperatures (up to 80°C), demonstrating that these consortia are an excellent source of enzymes for the development of enzymatic cocktails tailored to more extreme reaction conditions.
机译:工业规模的生物燃料生产需要强大的酶混合物来从木质纤维素生物质生产可发酵糖。嗜热细菌聚生体是纤维素酶和半纤维素酶的潜在来源,与商业真菌酶相比,纤维素酶和半纤维素酶适合于更苛刻的反应条件。来自堆肥的微生物聚生体适用于60°C的柳枝to,以开发嗜热生物质降解聚生体,以进行详细研究。使用小亚基rRNA基因扩增子焦磷酸测序和短读宏基因组测序进行的微生物群落分析表明,嗜热适应柳枝switch的结果是低多样性细菌聚生,其中包括与嗜热念珠菌,罗氏嗜热菌和嗜热栖热菌相关的大量细菌。在较低的丰度下,观察到嗜热的叶绿藻和未培养的Gemmatimonadetes phylum。从这些财团中分离出的上清液具有高水平的木聚糖酶和内切葡聚糖酶活性。与市售酶制剂相比,内切葡聚糖酶具有更高的耐热性,并且在存在用于生物质预处理的离子液体乙酸1-乙基-3-甲基咪唑鎓盐([C2mim] [OAc])时更稳定。上清液用于在高温(高达80°C)下糖化[C2mim] [OAc]预处理的柳枝,,证明这些聚生体是开发适合更极端反应条件的酶混合物的出色酶源。

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