WFRF:(Wang Yanyan 1989)
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- Wang, Yanyan,1989Chalmers tekniska högskola,Chalmers University of Technology (author)
CRISPR/Cas9-mediated point mutations improve alpha-amylase secretion in Saccharomyces cerevisiae
- Article/chapterEnglish2022
Publisher, publication year, extent ...
- 2022-07-01
- Oxford University Press (OUP),2022
- electronicrdacarrier
Numbers
- LIBRIS-ID:oai:research.chalmers.se:cdd78f81-2d6a-4cde-84b5-eb71bfc43cd4
- https://research.chalmers.se/publication/531366URI
- https://doi.org/10.1093/femsyr/foac033DOI
Supplementary language notes
- Language:English
- Summary in:English
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- The rapid expansion of the application of pharmaceutical proteins and industrial enzymes requires robust microbial workhorses for high protein production. The budding yeast Saccharomyces cerevisiae is an attractive cell factory due to its ability to perform eukaryotic post-translational modifications and to secrete proteins. Many strategies have been used to engineer yeast platform strains for higher protein secretion capacity. Herein, we investigated a line of strains that have previously been selected after UV random mutagenesis for improved alpha-amylase secretion. A total of 42 amino acid altering point mutations identified in this strain line were reintroduced into the parental strain AAC to study their individual effects on protein secretion. These point mutations included missense mutations (amino acid substitution), nonsense mutations (stop codon generation), and frameshift mutations. For comparison, single gene deletions for the corresponding target genes were also performed in this study. A total of 11 point mutations and seven gene deletions were found to effectively improve alpha-amylase secretion. These targets were involved in several bioprocesses, including cellular stresses, protein degradation, transportation, mRNA processing and export, DNA replication, and repair, which indicates that the improved protein secretion capacity in the evolved strains is the result of the interaction of multiple intracellular processes. Our findings will contribute to the construction of novel cell factories for recombinant protein secretion. Systematic characterization of point mutations from evolved strains using CRISPR/Cas9 technology revealed a set of gene alterations that improved recombinant protein secretion in Saccharomyces cerevisiae.
Subject headings and genre
- NATURVETENSKAP Biologi Cellbiologi hsv//swe
- NATURAL SCIENCES Biological Sciences Cell Biology hsv//eng
- NATURVETENSKAP Biologi Biokemi och molekylärbiologi hsv//swe
- NATURAL SCIENCES Biological Sciences Biochemistry and Molecular Biology hsv//eng
- MEDICIN OCH HÄLSOVETENSKAP Medicinsk bioteknologi Medicinsk bioteknologi hsv//swe
- MEDICAL AND HEALTH SCIENCES Medical Biotechnology Medical Biotechnology hsv//eng
- protein production
- Cas9
- gene deletion
- point mutation
- recombinant proteins
- CRISPR
Added entries (persons, corporate bodies, meetings, titles ...)
- Li, Xiaowei,1986Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)lixiaow (author)
- Chen, Xin,1980Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)cxin (author)
- Siewers, Verena,1976Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)siewers (author)
- Chalmers tekniska högskola (creator_code:org_t)
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- In:FEMS Yeast Research: Oxford University Press (OUP)22:11567-13561567-1364
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