4.8 Article

A universal glycoenzyme biosynthesis pipeline that enables efficient cell-free remodeling of glycans

期刊

NATURE COMMUNICATIONS
卷 13, 期 1, 页码 -

出版社

NATURE PORTFOLIO
DOI: 10.1038/s41467-022-34029-7

关键词

-

资金

  1. Bill and Melinda Gates Foundation [OPP1217652]
  2. Defense Threat Reduction Agency [HDTRA1-15-10052, HDTRA1-20-10004]
  3. National Science Foundation [CBET-1159581, CBET-1264701, CBET-1936823, MCB-1413563]
  4. National Institutes of Health [1R01GM137314, 1R01GM127578, R01GM130915, 1U54CA210184]
  5. Royal Thai Government Fellowship
  6. Cornell Fleming Graduate Scholarship
  7. Bill and Melinda Gates Foundation [OPP1217652] Funding Source: Bill and Melinda Gates Foundation

向作者/读者索取更多资源

This study describes a strategy to convert membrane-bound glycosyltransferases into water soluble biocatalysts, which allows for easier expression and structural remodeling of glycans. The approach was successfully used to produce 98 difficult-to-express GTs, predominantly of human origin, and demonstrated its universality and versatility.
The ability to reconstitute natural glycosylation pathways or prototype entirely new ones from scratch is hampered by the limited availability of functional glycoenzymes, many of which are membrane proteins that fail to express in heterologous hosts. Here, we describe a strategy for topologically converting membrane-bound glycosyltransferases (GTs) into water soluble biocatalysts, which are expressed at high levels in the cytoplasm of living cells with retention of biological activity. We demonstrate the universality of the approach through facile production of 98 difficult-to-express GTs, predominantly of human origin, across several commonly used expression platforms. Using a subset of these water-soluble enzymes, we perform structural remodeling of both free and protein-linked glycans including those found on the monoclonal antibody therapeutic trastuzumab. Overall, our strategy for rationally redesigning GTs provides an effective and versatile biosynthetic route to large quantities of diverse, enzymatically active GTs, which should find use in structure-function studies as well as in biochemical and biomedical applications involving complex glycomolecules.

作者

我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。

评论

主要评分

4.8
评分不足

次要评分

新颖性
-
重要性
-
科学严谨性
-
评价这篇论文

推荐

暂无数据
暂无数据