Secretome-based screening of fusion partners and their application in recombinant protein secretion in Saccharomyces cerevisiae

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Title
Secretome-based screening of fusion partners and their application in recombinant protein secretion in Saccharomyces cerevisiae
Author(s)
Jung Hoon Bae; S H Yun; Mi-Jin Kim; Hyun-Jin Kim; Bong Hyun Sung; S I Kim; Jung Hoon Sohn
Bibliographic Citation
Applied Microbiology and Biotechnology, vol. 106, no. 2, pp. 663-673
Publication Year
2022
Abstract
For the efficient production of heterologous proteins in the yeast Saccharomyces cerevisiae, we screened for a novel fusion partner from the yeast secretome. From twenty major proteins identified from the yeast secretome, we selected Scw4p, a cell wall protein with similarity to glucanase, and modified to develop a general fusion partner for the secretory expression of heterologous proteins in yeast. The optimal size of the SCW4 gene to act as an efficient fusion partner was determined by C-terminal truncation analysis; two of the variants, S1 (truncated at codon 115Q) and S2 (truncated at codon 142E), were further used for the secretion of heterologous proteins. When fused with S2, the secretion of three target proteins (hGH, exendin-4, and hPTH) significantly increased. Conserved O-glycosylation sites (Ser/Thr-rich domain) and hydrophilic sequences of S2 were deemed important for the function of S2 as a secretion fusion partner. Approximately 5 g/L of the S2-exendin-4 fusion protein was obtained from fed-batch fermentation. Intact target proteins were easily purified by affinity chromatography after in vitro processing of the fusion partner. This system may be of general application for the secretory production of heterologous proteins in S. cerevisiae. KEY POINTS : ? Target proteins were efficiently secreted with their N-terminus fused to Scw4p. ? O-glycosylation and hydrophilic stretches in Scw4p were important for protein secretion. ? A variant of Scw4p (S2) was successfully applied for the secretory expression of heterologous proteins.
Keyword
Recombinant protein productionSecretionSCW4Fusion partnerhGHExendin-4hPTH
ISSN
0175-7598
Publisher
Springer
DOI
http://dx.doi.org/10.1007/s00253-021-11750-9
Type
Article
Appears in Collections:
Synthetic Biology and Bioengineering Research Institute > Synthetic Biology Research Center > 1. Journal Articles
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