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

Overview

Journal J Microbiol Biotechnol

Specialties Biotechnology
Microbiology

Date 2023 Jul 24

PMID 37482809

Authors

Mi-Jin Kim

Se-Lin Park

Seung Hwa Kim

Hyun-Joo Park

Bong Hyun Sung

Jung-Hoon Sohn

Jung-Hoon Bae

Affiliations

Soon will be listed here.

Abstract

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 . 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, P', P', P, and P sites of Kex2p cleavage sites were elaborately manipulated. The general use of Kex2p in recombinant protein production was confirmed using several recombinant proteins.

Citing Articles

Functional expression of recombinant insulins in Saccharomyces cerevisiae.

Kim M, Park S, Kim H, Sung B, Sohn J, Bae J Microb Cell Fact. 2024; 23(1):302.

PMID: 39529045 PMC: 11552327. DOI: 10.1186/s12934-024-02571-2.

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