Engineering strategies for enhanced heterologous protein production by Saccharomyces cerevisiae

Abstract

Microbial proteins are promising substitutes for animal- and plant-based proteins. S. cerevisiae, a generally recognized as safe (GRAS) microorganism, has been frequently employed to generate heterologous proteins. However, constructing a universal yeast chassis for efficient protein production is still a challenge due to the varying properties of different proteins. With progress in synthetic biology, a multitude of molecular biology tools and metabolic engineering strategies have been employed to alleviate these issues. This review first analyses the advantages of protein production by S. cerevisiae. The most recent advances in improving heterologous protein yield are summarized and discussed in terms of protein hyperexpression systems, protein secretion engineering, glycosylation pathway engineering and systems metabolic engineering. Furthermore, the prospects for efficient and sustainable heterologous protein production by S. cerevisiae are also provided.

Keywords: Expression system; Protein production; Saccharomyces cerevisiae; Secretion engineering; Systems metabolic engineering.

Fig. 1

Construction of S. cerevisiae cell factories. The advent of new technologies has paved the way for designing S. cerevisiae to become a perfect production platform, significantly reducing strain construction time and accelerating the entire design, build, test, and learning cycle

Fig. 2

A review of engineering strategies for improved protein production by S. cerevisiae, including the construction of a hyperexpression system, secretion engineering, glycosylation pathway engineering, and systems metabolic engineering

Fig. 3

Strategies for protein hyperexpression systems, including codon optimization, increasing gene copy numbers, and transcriptional regulation

Fig. 4

Promoter engineering for protein production in S. cerevisiae. (A) Random mutation and screening of promoter libraries. (B) Construction of the minimal promoter construct. (C) Combination of each element for hybrid promoters. (D) Machine learning procedures for promoter design

Fig. 5

Protein secretion engineering in S. cerevisiae, including secretion signal engineering, ER folding engineering, and vesicle trafficking engineering

Fig. 6

The construction of a high-protein-producing yeast assisted by systems metabolic engineering, including improving substance and energy metabolism for protein synthesis, reducing oxidative stress, and rationally engineering metabolic pathways guided by multiomics data and constrained metabolic network models