Cell surface display of salmobin, a thrombin-like enzyme from Agkistrodon halys venom on Escherichia coli using ice nucleation protein

被引：27
作者：
Jeong, HS
Yoo, SK
Kim, EJ
机构：
[1] Yonsei Univ, Coll Engn, Dept Biotechnol, Seoul 120749, South Korea
[2] Yonsei Univ, Bioprod Res Ctr, Seoul 120749, South Korea
来源：
ENZYME AND MICROBIAL TECHNOLOGY | 2001年 / 28卷 / 2-3期
关键词：
cell surface display; ice nucleation protein; thrombin-like enzyme; salmobin;
D O I：
10.1016/S0141-0229(00)00315-X
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：
Cell surface display on Escherichia coli using ice nucleation protein was performed in order to develop a new expression system for recombinant eukaryotic proteins. Salmobin, the thrombin-like enzyme obtained from Korean snake (Agkistrodon halys) venom was displayed on the surface of Escherichia coli fused to the C-terminus of the ice nucleation protein (INP). an outer membrane protein of Pseudomonas syringae. The thrombin cleavage site was inserted between salmobin and INP. The presence of salmobin on the bacterial cell surface was verified by SDS-PAGE, Western blotting, whole cell ELISA, and immunofluorescence microscopy. After thrombin cleavage the thrombin-like enzyme activity of recombinant salmobin was tested and verified. We concluded that INP-based cell surface display can be used as a novel expression system for eukaryotic proteins. (C) 2001 Elsevier Science Inc. All rights reserved.
引用
收藏
页码：155 / 160
页数：6
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