ArticleBiotechnology for biofuels2012
PGASO: A synthetic biology tool for engineering a cellulolytic yeast.
Article in Biotechnology for biofuels, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed, 49 citations in OpenAlex.
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- Growth and autolysis of the kefir yeast Kluyveromyces marxianus in lactate culture.Scientific reports · 2021Article
- Heterologous Expression of Lignocellulose-Modifying Enzymes in Microorganisms: Current Status.Molecular biotechnology · 2021Review
- Construction of engineered RuBisCO Kluyveromyces marxianus for a dual microbial bioethanol production system.PloS one · 2021Article
- Constructing a yeast to express the largest cellulosome complex on the cell surface.Proceedings of the National Academy of Sciences of the United States of America · 2020Article
- Characterizing an engineered carotenoid-producing yeast as an anti-stress chassis for building cell factories.Microbial cell factories · 2019Article
- Biological Parts forFrontiers in bioengineering and biotechnology · 2019Article
- Engineering the oleaginous red yeast Rhodotorula glutinis for simultaneous β-carotene and cellulase production.Scientific reports · 2018Article
- Applications of Yeast Synthetic Biology Geared towards the Production of Biopharmaceuticals.Genes · 2018Review
- Genome-wide prediction of CRISPR/Cas9 targets in Kluyveromyces marxianus and its application to obtain a stable haploid strain.Scientific reports · 2018Article
- Construction of a cellulose-metabolizing Komagataella phaffii (Pichia pastoris) by co-expressing glucanases and β-glucosidase.Applied microbiology and biotechnology · 2018Article
- Biochemical characterization and structural analysis of a bifunctional cellulase/xylanase from Clostridium thermocellum.The Journal of biological chemistry · 2015Article
- Challenges and advances in the heterologous expression of cellulolytic enzymes: a review.Biotechnology for biofuels · 2014Article
- Assembling a cellulase cocktail and a cellodextrin transporter into a yeast host for CBP ethanol production.Biotechnology for biofuels · 2013Article
Corrections and comments
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Authors and funding
10 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundTo achieve an economical cellulosic ethanol production, a host that can do both cellulosic saccharification and ethanol fermentation is desirable. However, to engineer a non-cellulolytic yeast to be such a host requires synthetic biology techniques to transform multiple enzyme genes into its genome.
resultsA technique, named Promoter-based Gene Assembly and Simultaneous Overexpression (PGASO), that employs overlapping oligonucleotides for recombinatorial assembly of gene cassettes with individual promoters, was developed. PGASO was applied to engineer Kluyveromycesmarxianus KY3, which is a thermo- and toxin-tolerant yeast. We obtained a recombinant strain, called KR5, that is capable of simultaneously expressing exoglucanase and endoglucanase (both of Trichodermareesei), a beta-glucosidase (from a cow rumen fungus), a neomycin phosphotransferase, and a green fluorescent protein. High transformation efficiency and accuracy were achieved as ~63% of the transformants was confirmed to be correct. KR5 can utilize beta-glycan, cellobiose or CMC as the sole carbon source for growth and can directly convert cellobiose and beta-glycan to ethanol.
conclusionsThis study provides the first example of multi-gene assembly in a single step in a yeast species other than Saccharomyces cerevisiae. We successfully engineered a yeast host with a five-gene cassette assembly and the new host is capable of co-expressing three types of cellulase genes. Our study shows that PGASO is an efficient tool for simultaneous expression of multiple enzymes in the kefir yeast KY3 and that KY3 can serve as a host for developing synthetic biology tools.
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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.