...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biochemistry >Overall Retention of Methyl Stereochemistry during B-12-Dependent Radical SAM Methyl Transfer in Fosfomycin Biosynthesis
【24h】

Overall Retention of Methyl Stereochemistry during B-12-Dependent Radical SAM Methyl Transfer in Fosfomycin Biosynthesis

机译：在FOSFOMYCIN生物合成中B-12依赖性自由基SAM甲基转移过程中甲基立体化学的总体抑制

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

Methylcobalamin-dependent radical S-adenosylmethionine (SAM) enzymes methylate non-nucleophilic atoms in a range of substrates. The mechanism of the methyl transfer from cobalt to the receiving atom is still mostly unresolved. Here we determine the stereochemical course of this process at the methyl group during the biosynthesis of the clinically used antibiotic fosfomycin. In vitro reaction of the methyltransferase Fom3 using SAM labeled with H-1, H-2, and H-3 in a stereochemically defined manner, followed by chemoenzymatic conversion of the Fom3 product to acetate and subsequent stereochemical analysis, shows that the overall reaction occurs with retention of configuration. This outcome is consistent with a double-inversion process, first in the S(N)2 reaction of cob(I)alamin with SAM to form methylcobalamin and again in a radical transfer of the methyl group from methylcobalamin to the substrate. The methods developed during this study allow high-yield in situ generation of labeled SAM and recombinant expression and purification of the malate synthase needed for chiral methyl analysis. These methods facilitate the broader use of in vitro chiral methyl analysis techniques to investigate the mechanisms of other novel enzymes.
机译：Methylcobalamin-dependent radical s-adenosylmethionine（SAM）enzymes methylate non-nucleophilic atoms in a range of substrates.从Cobalt到原子接收的方法转换的机制仍然没有得到解决。在这里，我们定义了这一过程的立体化学课程在Methyl集团在临床使用的抗菌fosfomycin的生物合成过程中。在体视反应中，Methyltransferase FOM3使用与H-1、H-2和H-3在一个立体化学定义的manner中的SAM实验室，遵循FOM3产品的化学转化为acetate和subsequent立体化学分析，显示超反应Occurs具有还原配置。这一结果是与一个双重渗透过程，首先在S（n）2反应COB（i）alamin与SAM的形式methylcobalamin，并再次在一个从methylcobalamin到substrate的辐射转移Methyl组。The methods developed during this study allow high-yield in situ generation of labeled sam and recombinant expression and purification of the malate synthase need for chiral methyl analysis.这些方法有助于广告商使用体液化学分析技术来调查其他小说谜团的机制。

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