Wang Guozeng, Huang Xiaoyun, Ng Tzi Bun, Lin Juan, Ye Xiu Yun
College of Biological Science and Technology, Fuzhou University, Fuzhou 350108, P.R. China; National Engineering Laboratory for High-efficiency Enzyme Expression, Fuzhou 350002, P. R. China.
College of Biological Science and Technology, Fuzhou University, Fuzhou 350108, P.R. China.
PLoS One. 2014 Nov 13;9(11):e112798. doi: 10.1371/journal.pone.0112798. eCollection 2014.
Soda lakes are one of the most stable naturally occurring alkaline and saline environments, which harbor abundant microorganisms with diverse functions. In this study, culture-independent molecular methods were used to explore the genetic diversity of glycoside hydrolase (GH) family 10 and GH11 xylanases in Lake Dabusu, a soda lake with a pH value of 10.2 and salinity of 10.1%. A total of 671 xylanase gene fragments were obtained, representing 78 distinct GH10 and 28 GH11 gene fragments respectively, with most of them having low homology with known sequences. Phylogenetic analysis revealed that the GH10 xylanase sequences mainly belonged to Bacteroidetes, Proteobacteria, Actinobacteria, Firmicutes and Verrucomicrobia, while the GH11 sequences mainly consisted of Actinobacteria, Firmicutes and Fungi. A full-length GH10 xylanase gene (xynAS10-66) was directly cloned and expressed in Escherichia coli, and the recombinant enzymes showed high activity at alkaline pH. These results suggest that xylanase gene diversity within Lake Dabusu is high and that most of the identified genes might be novel, indicating great potential for applications in industry and agriculture.
苏打湖是最稳定的天然碱性和盐性环境之一,其中蕴藏着功能多样的丰富微生物。在本研究中,采用非培养分子方法探究了达布苏湖(一个pH值为10.2、盐度为10.1%的苏打湖)中糖苷水解酶(GH)家族10和GH11木聚糖酶的遗传多样性。共获得671个木聚糖酶基因片段,分别代表78个不同的GH10和28个GH11基因片段,其中大多数与已知序列的同源性较低。系统发育分析表明,GH10木聚糖酶序列主要属于拟杆菌门、变形菌门、放线菌门、厚壁菌门和疣微菌门,而GH11序列主要由放线菌门、厚壁菌门和真菌组成。一个全长GH10木聚糖酶基因(xynAS10-66)被直接克隆并在大肠杆菌中表达,重组酶在碱性pH下显示出高活性。这些结果表明,达布苏湖内木聚糖酶基因多样性很高,且大多数已鉴定的基因可能是新的,这表明其在工农业应用中具有巨大潜力。