Modulation of Kex2p Cleavage Site for In Vitro Processing of Recombinant Proteins Produced by Saccharomyces cerevisiae

被引：2

作者：

Kim, Mi-Jin ^{[1
]}

Park, Se -Lin ^{[1
,2
]}

Kim, Seung Hwa ^{[1
,3
]}

Park, Hyun-Joo ^{[4
]}

Sung, Bong Hyun ^{[1
,3
]}

Sohn, Jung-Hoon ^{[1
,3
,4
]}

Bae, Jung-Hoon ^{[1
]}

机构：

[1] Korea Res Inst Biosci & Biotechnol, Synthet Biol Res Ctr, Daejeon 34141, South Korea

[2] Chungnam Natl Univ, Dept Food Sci & Technol, Daejeon 34134, South Korea

[3] Korea Univ Sci & Technol UST, KRIBB Sch Biotechnol, Dept Biosyst & Bioengn, Daejeon 34113, South Korea

[4] Cellapy Bio Inc, Bioventure Ctr 211, Daejeon 34141, South Korea

来源：

JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY | 2023年 / 33卷 / 11期

基金：

新加坡国家研究基金会;

关键词：

Recombinant protein; secretion; Kex2p; in-vitro processing; Saccharomyces cerevisiae; MOLECULAR-CLONING; FUSION PROTEINS; PROTEASE; SPECIFICITY; ENZYME; EXPRESSION; PRECURSOR; ENDOPEPTIDASE; SUBSITES; REVEALS;

D O I：

10.4014/jmb.2306.06024

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：

Kex2 protease (Kex2p) is a membrane-bound serine protease responsible for the proteolytic maturation of various secretory proteins by cleaving after dibasic residues in the late Golgi network. In this study, we present an application of Kex2p as an alternative endoprotease for the in vitro processing of recombinant fusion proteins produced by the yeast Saccharomyces cerevisiae. The proteins were expressed with a fusion partner connected by a Kex2p cleavage sequence for enhanced expression and easy purification. To avoid in vivo processing of fusion proteins by Kex2p during secretion and to guarantee efficient removal of the fusion partners by in vitro Kex2p processing, P1 ', P2 ', P4, and P3 sites of Kex2p cleavage sites were elaborately manipulated. The general use of Kex2p in recombinant protein production was confirmed using several recombinant proteins.

引用

页码：1513 / 1520

页数：8

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