ArticleApplied and environmental microbiology2014
Improved production of a heterologous amylase in Saccharomyces cerevisiae by inverse metabolic engineering.
Article in Applied and environmental microbiology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed, 37 citations in OpenAlex.
- Engineering strategies for enhanced heterologous protein production by Saccharomyces cerevisiae.Microbial cell factories · 2024Review
- Key anti-freeze genes and pathways of Lanzhou lily (Lilium davidii, var. unicolor) during the seedling stage.PloS one · 2024Article
- Strategies for the Development of Industrial Fungal Producing Strains.Journal of fungi (Basel, Switzerland) · 2023Review
- Improvement of cell-tethered cellulase activity in recombinant strains of Saccharomyces cerevisiae.Applied microbiology and biotechnology · 2022Article
- CRISPR/Cas9-mediated point mutations improve α-amylase secretion in Saccharomyces cerevisiae.FEMS yeast research · 2022Article
- A multistrategy approach for improving the expression of E. coli phytase in Pichia pastoris.Journal of industrial microbiology & biotechnology · 2020Article
- Engineering the protein secretory pathway ofProceedings of the National Academy of Sciences of the United States of America · 2018Article
- Microfluidic screening and whole-genome sequencing identifies mutations associated with improved protein secretion by yeast.Proceedings of the National Academy of Sciences of the United States of America · 2015Article
- Enhancement of phenol biodegradation by Pseudochrobactrum sp. through ultraviolet-induced mutation.International journal of molecular sciences · 2015Article
- Recent advances in combinatorial biosynthesis for drug discovery.Drug design, development and therapy · 2015Review
Corrections and comments
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Authors and funding
9 authors at 4 institutions in 2 countries.
Funding
Abstract
The increasing demand for industrial enzymes and biopharmaceutical proteins relies on robust production hosts with high protein yield and productivity. Being one of the best-studied model organisms and capable of performing posttranslational modifications, the yeast Saccharomyces cerevisiae is widely used as a cell factory for recombinant protein production. However, many recombinant proteins are produced at only 1% (or less) of the theoretical capacity due to the complexity of the secretory pathway, which has not been fully exploited. In this study, we applied the concept of inverse metabolic engineering to identify novel targets for improving protein secretion. Screening that combined UV-random mutagenesis and selection for growth on starch was performed to find mutant strains producing heterologous amylase 5-fold above the level produced by the reference strain. Genomic mutations that could be associated with higher amylase secretion were identified through whole-genome sequencing. Several single-point mutations, including an S196I point mutation in the VTA1 gene coding for a protein involved in vacuolar sorting, were evaluated by introducing these to the starting strain. By applying this modification alone, the amylase secretion could be improved by 35%. As a complement to the identification of genomic variants, transcriptome analysis was also performed in order to understand on a global level the transcriptional changes associated with the improved amylase production caused by UV mutagenesis.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.