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利用酶活性测定和基因组分析相结合的方法高效筛选博斯氏菌FBZP-16产生的潜在纤维素酶和半纤维素酶。

Efficient screening of potential cellulases and hemicellulases produced by Bosea sp. FBZP-16 using the combination of enzyme assays and genome analysis.

作者信息

Houfani Aicha Asma, Větrovský Tomáš, Baldrian Petr, Benallaoua Said

机构信息

Laboratoire de Microbiologie Appliquée (LMA), Département de Microbiologie, Faculté des Sciences de la Nature et de la Vie, Université de Bejaia, 06000, Bejaia, Algeria.

Laboratory of Environmental Microbiology, Institute of Microbiology of the Czech Academy of Sciences (CAS), Videnska 1083, 14220, Praha 4, Czech Republic.

出版信息

World J Microbiol Biotechnol. 2017 Feb;33(2):29. doi: 10.1007/s11274-016-2198-x. Epub 2017 Jan 5.

Abstract

Identification of bacteria that produce carbohydrolytic enzymes is extremely important given the increased demand for these enzymes in many industries. Twenty lignocellulose-degrading bacterial isolates from Algerian compost and different soils were screened for their potential to produce different enzymes involved in biomass deconstruction. Based on 16S rRNA gene sequencing, the isolates belonged to Proteobacteria and Actinobacteria. Differences among species were reflected both as the presence/absence of enzymes or at the level of enzyme activity. Among the most active species, Bosea sp. FBZP-16 demonstrated cellulolytic activity on both amorphous cellulose (CMC) and complex lignocellulose (wheat straw) and was selected for whole-genomic sequencing. The genome sequencing revealed the presence of a complex enzymatic machinery required for organic matter decomposition. Analysis of the enzyme-encoding genes indicated that multiple genes for endoglucanase, xylanase, β-glucosidase and β-mannosidase are present in the genome with enzyme activities displayed by the bacterium, while other enzymes, such as certain cellobiohydrolases, were not detected at the genomic level. This indicates that a combination of functional screening of bacterial cultures with the use of genome-derived information is important for the prediction of potential enzyme production. These results provide insight into their possible exploitation for the production of fuels and chemicals derived from plant biomass.

摘要

鉴于许多行业对碳水化合物水解酶的需求不断增加,鉴定产生碳水化合物水解酶的细菌极为重要。对从阿尔及利亚堆肥和不同土壤中分离出的20株木质纤维素降解细菌进行了筛选,以评估它们产生参与生物质解构的不同酶的潜力。基于16S rRNA基因测序,这些分离株属于变形菌门和放线菌门。物种间的差异既体现在酶的有无上,也体现在酶活性水平上。在最具活性的物种中,博斯氏菌属FBZP-16对无定形纤维素(羧甲基纤维素)和复杂木质纤维素(小麦秸秆)均表现出纤维素分解活性,并被选用于全基因组测序。基因组测序揭示了存在有机物分解所需的复杂酶系。对编码酶的基因分析表明,基因组中存在多个编码内切葡聚糖酶、木聚糖酶、β-葡萄糖苷酶和β-甘露糖苷酶的基因,且该细菌表现出这些酶的活性,而其他酶,如某些纤维二糖水解酶,在基因组水平未被检测到。这表明将细菌培养物的功能筛选与利用基因组衍生信息相结合,对于预测潜在的酶产量很重要。这些结果为它们在生产源自植物生物质的燃料和化学品方面的可能应用提供了见解。

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