• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Applied and Environmental Microbiology >P1 Trisaccharide (Galα1,4Galβ1,4GlcNAc) Synthesis by Enzyme Glycosylation Reactions Using Recombinant Escherichia coli
【24h】

P1 Trisaccharide (Galα1,4Galβ1,4GlcNAc) Synthesis by Enzyme Glycosylation Reactions Using Recombinant Escherichia coli

机译：重组大肠杆菌酶糖基化反应合成P1三糖（Galα1,4Galβ1,4GlcNAc）

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The frequency of Escherichia coli infection has lead to concerns over pathogenic bacteria in our food supply and a demand for therapeutics. Glycolipids on gut cells serve as receptors for the Shiga-like toxin produced by E. coli. Oligosaccharide moiety analogues of these glycolipids can compete with receptors for the toxin, thus acting as antibacterials. An enzymatic synthesis of the P1 trisaccharide (Galα1,4Galβ1,4GlcNAc), one of the oligosaccharide analogues, was assessed in this study. In the proposed synthetic pathway, UDP-glucose was generated from sucrose with an Anabaena sp. sucrose synthase and then converted with an E. coli UDP-glucose 4-epimerase to UDP-galactose. Two molecules of galactose were linked to N-acetylglucosamine subsequently with a Helicobacter pylori β-l,4-galactosyltransferase and a Neisseria meningitidis α-1,4-galactosyltransferase to produce one molecule of P1 trisaccharide. The four enzymes were coexpressed in a single genetically engineered E. coli strain that was then permeabilized and used to catalyze the enzymatic reaction. P1 trisaccharide was accumulated up to 50 mM (5.4 g in a 200-ml reaction volume), with a 67% yield based on the consumption of N-acetylglucosamine. This study provides an efficient approach for the preparative-scale synthesis of P1 trisaccharide with recombinant bacteria.
机译：大肠埃希氏菌感染的频率已引起人们对我们食物供应中的病原菌和对治疗剂的需求的担忧。肠道细胞上的糖脂可作为大肠杆菌产生的志贺氏样毒素的受体。这些糖脂的寡糖部分类似物可与毒素竞争受体，从而起到抗菌作用。这项研究评估了低聚糖类似物之一的P1三糖（Galα1,4Galβ1,4GlcNAc）的酶促合成。在拟议的合成途径中，UDP和葡萄糖是由蔗糖和鱼腥藻产生的。蔗糖合酶，然后用大肠杆菌UDP-葡萄糖4-表异构酶转化为UDP-半乳糖。随后用幽门螺杆菌β-1,4-半乳糖基转移酶和脑膜炎奈瑟氏球菌α-1,4-半乳糖基转移酶将两个分子的半乳糖与N-乙酰氨基葡萄糖连接，以产生一分子的P1三糖。四种酶在单个基因工程大肠杆菌菌株中共表达，然后被渗透并用于催化酶促反应。 P1三糖的累积量高达50 mM（在200 ml反应体积中为5.4 g），基于N-乙酰氨基葡萄糖的消耗，收率为67％。这项研究为重组细菌制备规模规模的P1三糖提供了一种有效的方法。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
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