期刊
CARBOHYDRATE POLYMERS
卷 278, 期 -, 页码 -出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.carbpol.2021.118951
关键词
Paenibacillus polymyxa; Paenan; CRISPR-Cas9; Exopolysaccharides; Fucose
资金
- German Federal Ministry of Education and Research (BMBF) [03VP02560]
- Research Council of Norway [226244, 294946]
By using the CRISPR-Cas9 mediated knock-out strategy, a pure fucose-containing polysaccharide named paenan II was successfully isolated from Paenibacillus polymyxa, and its structure was determined using various analytical methods.
Paenibacillus polymyxa is an avid producer of exopolysaccharides of industrial interest. However, due to the complexity of the polymer composition, structural elucidation of the polysaccharide remained unfeasible for a long time. By using a CRISPR-Cas9 mediated knock-out strategy, all single glycosyltransferases as well as the Wzy polymerases were individually deleted in the corresponding gene cluster for the first time. Thereby, it was observed that the main polymer fraction was completely suppressed (or deleted) and a pure minor fucose containing polysaccharide could be isolated, which was named paenan II. Applying this combinatorial approach, the monosaccharide composition, sequence and linkage pattern of this novel polymer was determined via HPLC-MS, GC-MS and NMR. Furthermore, we demonstrated that the knock-out of the glycosyltransferases PepQ, PepT, PepU and PepV as well as of the Wzy polymerase PepG led to the absence of paenan II, attributing those enzymes to the assembly of the repeating unit.
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。
推荐
暂无数据