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从堆肥宏基因组中获得的新GH43基因具有双木聚糖酶和纤维素酶活性。

Newly derived GH43 gene from compost metagenome showing dual xylanase and cellulase activities.

作者信息

Sae-Lee Ritthironk, Boonmee Atcha

机构信息

Department of Microbiology, Faculty of Science, Khon Kaen University, Khon Kaen, 40002, Thailand.

出版信息

Folia Microbiol (Praha). 2014 Sep;59(5):409-17. doi: 10.1007/s12223-014-0313-7. Epub 2014 Apr 16.

Abstract

A metagenomic fosmid library was constructed from compost microbial communities that were collected from various farms throughout the Khon Kaen province, Thailand. The library was enriched in carboxymethylcellulose (CM-cellulose)--containing media prior to the screening of clones capable of degrading cellulosic materials. Two clones were selected for further subcloning and sequencing based on different patterns from restriction analysis. Deduced amino acid analysis of possible ORFs revealed one novel gene encoding an enzyme belonging to glycosyl hydrolase family 43 (GH43), which is a GH family rarely found in metagenomic studies. The most notable finding is that this enzyme, designated as Biof1_09, shows dual activities, namely endocellulase and endoxylanase activities. Biof1_09 showed greater than 50% of its activity under acidic conditions ranging from pH 3.5 to 5.5 with a pH optimum of 4.5. The optimum temperature of this enzyme was between 45 and 55 °C with an optimum of 50 °C. The properties of Biof1_09 make this enzyme an attractive candidate for large-scale expression for use in lignocellulose degradation for various bioprocess applications, including bioethanol fermentation.

摘要

从泰国孔敬府各地农场收集的堆肥微生物群落构建了一个宏基因组fosmid文库。在筛选能够降解纤维素材料的克隆之前,该文库在含有羧甲基纤维素(CM-纤维素)的培养基中进行了富集。根据限制性分析的不同模式,选择了两个克隆进行进一步的亚克隆和测序。对可能的开放阅读框进行推导氨基酸分析,发现了一个编码属于糖基水解酶家族43(GH43)的酶的新基因,这是一个在宏基因组研究中很少发现的GH家族。最值得注意的发现是,这种酶命名为Biof1_09,具有双重活性,即内切纤维素酶和内切木聚糖酶活性。Biof1_09在pH 3.5至5.5的酸性条件下显示出超过50%的活性,最适pH为4.5。该酶的最适温度在45至55℃之间,最适温度为50℃。Biof1_09的特性使其成为大规模表达的有吸引力的候选酶,可用于各种生物工艺应用中的木质纤维素降解,包括生物乙醇发酵。

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