Yeast cells harboring human α-1,3-fucosyltransferase at the cell surface engineered using Pir, a cell wall-anchored protein

被引：21

作者：

Abe, H ^{[1
]}

Ohba, M ^{[1
]}

Shimma, Y ^{[1
]}

Jigami, Y ^{[1
]}

机构：

[1] AIST, RCG, Tsukuba, Ibaraki 3058566, Japan

来源：

FEMS YEAST RESEARCH | 2004年 / 4卷 / 4-5期

关键词：

fucosyltransferase; intact cell enzyme; oligosaccharide synthesis; Pit; Saccharomyces cerevisiae;

D O I：

10.1016/S1567-1356(03)00193-4

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Human alpha-1,3-fucosyltansferase (FucT) encoded by the FUT6 gene was displayed at the cell surface of yeast cells engineered using the yeast cell wall protein Pir1 or Pir2, and the FucT activity was detected at the surface of cells producing the Pir1-HA-FUT6 or Pir2-FLAG-FUT6 fusion proteins. To obtain higher activity, we engineered the host yeast cells in which endogenous PIR genes of the PIR1-4 gene family were disrupted. Among the disruptants, the pir1Delta pir2Delta pir3Delta strain with the PIR1-HA-FUT6 fusion gene showed the highest FucT activity, which was about three-fold higher than that of the wild-type strain. Furthermore, the co-expression of both the Pir1-HA-FUT6 and the Pir2-FLAG-FUT6 fusions showed an approximately 1.5-fold higher activity than that in the cell wall displaying Pir1-HA-FUT6 alone. The present method was thus effective for producing yeast cells that can easily synthesize various oligosaccharides, such as Le(x) and sLe(x), using Pir-glycosyltransferase fusions in combination with the deletion of endogenous PIR genes. (C) 2003 Published by Elsevier B.V. on behalf of the Federation of European Microbiological Societies.

引用

页码：417 / 425

页数：9

共 32 条

[1] In vitro oligosaccharide synthesis using intact yeast cells that display glycosyltransferases at the cell surface through cell wall-anchored protein Pir [J].