...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Applied Microbiology and Biotechnology >Diversity of hydrolases from hydrothermal vent sediments of the Levante Bay, Vulcano Island (Aeolian archipelago) identified by activity-based metagenomics and biochemical characterization of new esterases and an arabinopyranosidase
【24h】

Diversity of hydrolases from hydrothermal vent sediments of the Levante Bay, Vulcano Island (Aeolian archipelago) identified by activity-based metagenomics and biochemical characterization of new esterases and an arabinopyranosidase

机译：来自伏尔加诺岛（风群岛）莱万特湾热液喷口沉积物的水解酶多样性通过基于活性的宏基因组学和新的酯酶和阿拉伯吡喃糖苷酶的生化特性鉴定

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

A metagenomic fosmid expression library established from environmental DNA (eDNA) from the shallow hot vent sediment sample collected from the Levante Bay, Vulcano Island (Aeolian archipelago) was established in Escherichia coli. Using activity-based screening assays, we have assessed 9600 fosmid clones corresponding to approximately 350 Mbp of the cloned eDNA, for the lipases/esterases/lactamases, haloalkane and haloacid dehalogenases, and glycoside hydrolases. Thirty-four positive fosmid clones were selected from the total of 120 positive hits and sequenced to yield ca. 1360 kbp of high-quality assemblies. Fosmid inserts were attributed to the members of ten bacterial phyla, including Proteobacteria, Bacteroidetes, Acidobateria, Firmicutes, Verrucomicrobia, Chloroflexi, Spirochaetes, Thermotogae, Armatimonadetes, and Planctomycetes. Of ca. 200 proteins with high biotechnological potential identified therein, we have characterized in detail three distinct alpha/beta-hydrolases (LIPESV12_9, LIPESV12_24, LIPESV12_26) and one new alpha-arabinopyranosidase (GLV12_5). All LIPESV12 enzymes revealed distinct substrate specificities tested against 43 structurally diverse esters and 4 p-nitrophenol carboxyl esters. Of 16 different glycosides tested, the GLV12_5 hydrolysed only p-nitrophenol-alpha-(l)-arabinopyranose with a high specific activity of about 2.7 kU/mg protein. Most of the alpha/beta-hydrolases were thermophilic and revealed a high tolerance to, and high activities in the presence of, numerous heavy metal ions. Among them, the LIPESV12_24 was the best temperature-adapted, retaining its activity after 40 min of incubation at 90 A degrees C. Furthermore, enzymes were active in organic solvents (e.g., > 30 % methanol). Both LIPESV12_24 and LIPESV12_26 had the GXSXG pentapeptides and the catalytic triads Ser-Asp-His typical to the representatives of carboxylesterases of EC 3.1.1.1.
机译：在大肠杆菌中建立了一个宏基因组化的fosmid表达文库，该文库由环境DNA（eDNA）建立，该文库来自于从伏尔卡诺岛（风群岛）的莱万特湾收集的浅热喷口沉积物样品。使用基于活性的筛选分析，我们评估了9600个fosmid克隆，它们对应于约350 Mbp的克隆eDNA，用于脂肪酶/酯酶/内酰胺酶，卤代烷烃和卤酸脱卤酶以及糖苷水解酶。从总共120个阳性结果中选择了34个阳性fosmid阳性克隆，并进行了测序，结果大约为。 1360 kbp的高质量装配体。 fosmid插入物归因于十种细菌门的成员，包括变形杆菌，拟杆菌，嗜酸细菌，硬毛菌，疣状微生物，绿弯曲菌，螺旋体，嗜热菌，腕足动物和扁平菌。约。在其中鉴定出200种具有高生物技术潜力的蛋白质中，我们已详细表征了三种不同的α/β水解酶（LIPESV12_9，LIPESV12_24，LIPESV12_26）和一种新的α-阿拉伯吡喃糖苷酶（GLV12_5）。所有LIPESV12酶均显示出针对43种结构多样的酯和4种对硝基苯酚羧基酯的独特底物特异性。在测试的16种不同的糖苷中，GLV12_5仅水解对硝基苯酚-α-（l）-阿拉伯吡喃糖，具有约2.7 kU / mg蛋白的高比活性。大多数α/β水解酶是嗜热的，并且在存在多种重金属离子时显示出高度的耐受性和高活性。其中，LIPESV12_24具有最佳的温度适应性，在90 A的温度下孵育40分钟后仍保持其活性。此外，酶在有机溶剂（例如> 30％的甲醇）中具有活性。 LIPESV12_24和LIPESV12_26均具有GXSXG五肽和EC 3.1.1.1羧酸酯酶代表典型的催化三联体Ser-Asp-His。

著录项

相似文献

  • 外文文献
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号