Comprehensive Analysis of Signal Peptides in Saccharomyces cerevisiae Reveals Features for Efficient Secretion

Abstract

Signal peptides (SPs) are N‐terminus sequences on the nascent polypeptide for protein export or localization delivery, which are essential for maintaining cell function. SPs are also employed as a key element for industrial production of secreted recombinant proteins. Yet, detailed information and rules about SPs and their cellular interactions are still not well understood. Here, systematic bioinformatics analysis and secretion capacity measurement of genome‐wide SPs from the model organism Saccharomyces cerevisiae is performed. Several key features of SPs, including region properties, consensus motifs, evolutionary relationships, codon bias, e.g., are successfully revealed. Diverse cell metabolism can be trigged by using different SPs for heterologous protein secretion. Influences on SPs with different properties by chaperones can cause different secretory efficiencies. Protein secretion by the SP NCW2 in SEC72 deletion strain is 10 times than the control. These findings provide insights into the properties and functions of SPs and contribute to both fundamental research and industrial application.

Exploiting Systematic Engineering of the Expression Cassette as a Powerful Tool to Enhance Heterologous Gene Expression in Trichoderma reesei

“…Moreover, these promoters were also reported to be able to drive heterologous gene expression in T. reesei. ,, As we previously have engineered the cbh1 promoter to pCbh1pX (Table S2), which has an improved efficiency to drive transcription, we included the pCbh1pX promoter in the comparison, as well.…”

“…In eukaryotic cells, a signal peptide directs the secretion of a nascent protein by translocating across the endoplasmic reticulum and Golgi apparatus for ultimate release into the extracellular milieu. In yeasts, filamentous fungi, and mammalian cells, changing the signal peptide can largely alter the expression level of a protein. , This implies that efficient secretion of a protein of interest requires fine-tuned compatibility of the signal peptide with the protein translocation machinery, although the underlying mechanisms are still not thoroughly understood.…”

“…In yeasts, filamentous fungi, and mammalian cells, changing the signal peptide can largely alter the expression level of a protein. 28,31 This implies that efficient secretion of a protein of interest requires fine-tuned compatibility of the signal peptide with the protein translocation machinery, although the underlying mechanisms are still not thoroughly understood.…”

Optimization of Protein Folding for Improved Secretion of Human Serum Albumin Fusion Proteins in Saccharomyces cerevisiae

“…cerevisiae are largely produced during the ethanol consumption phase. Meanwhile, glycerol and acetate levels offer insights into the metabolic activity and energy status of the yeast cells. ,,, To investigate the metabolic characteristics and production processes of the amylase-HSA fusion protein, samples were taken from cultures of strains C0 (harboring the empty plasmid CPOTud), AH0, and AH28 at different time points (Figure ). Compared with strain C0, strain AH0 containing plasmid pCP-AGSH exhibited slower growth and lower final biomass (Figure B).…”

The signal sequence of yeast killer toxin K2 confers producer self-protection and allows conversion into a modular toxin-antitoxin system

“…Given K2 is a secreted protein and thus requires a signal sequence for secretion, we manually searched the N-terminus for known features of a signal peptide. Signal sequences in S. cerevisiae are typically 18-24 residues long, and they feature a hydrophobic alpha-helical region (H-region), flanked by a positively charged N-terminal region (N-region) and a C-terminal region (C-region) that potentially contains a recognition sequence for the signal peptidase 21 ( Figure 2B ) .…”