ArticleApplied and environmental microbiology2013
Combinatorial design of a highly efficient xylose-utilizing pathway in Saccharomyces cerevisiae for the production of cellulosic biofuels.
Article in Applied and environmental microbiology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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29 citing papers in PubMed, 82 citations in OpenAlex.
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- Dynamic modulation of enzyme activity by synthetic CRISPR-Cas6 endonucleases.Nature chemical biology · 2022Article
- Fine-tuning the expression of pathway gene in yeast using a regulatory library formed by fusing a synthetic minimal promoter with different Kozak variants.Microbial cell factories · 2021Article
- Genetic circuit characterization by inferring RNA polymerase movement and ribosome usage.Nature communications · 2020Article
- Metabolic engineering considerations for the heterologous expression of xylose-catabolic pathways in Saccharomyces cerevisiae.PloS one · 2020Article
- Pentose Metabolism in Saccharomyces cerevisiae: The Need to Engineer Global Regulatory Systems.Biotechnology journal · 2019Review
- Applications of Yeast Synthetic Biology Geared towards the Production of Biopharmaceuticals.Genes · 2018Review
- Condition-specific promoter activities in Saccharomyces cerevisiae.Microbial cell factories · 2018Article
- Genomic and phenotypic characterization of a refactored xylose-utilizingBiotechnology for biofuels · 2018Article
- Engineering biological systems using automated biofoundries.Metabolic engineering · 2017Review
- Engineered yeast with a COScientific reports · 2017Article
- Systematic Optimization of Protein Secretory Pathways inFrontiers in microbiology · 2017Article
- High-throughput evaluation of synthetic metabolic pathways.Technology · 2016Article
- Metabolic engineering of Saccharomyces cerevisiae to produce 1-hexadecanol from xylose.Microbial cell factories · 2016Article
- Directed evolution of a fungal β-glucosidase in Saccharomyces cerevisiae.Biotechnology for biofuels · 2016Article
- Irradiation of Yarrowia lipolytica NRRL YB-567 creating novel strains with enhanced ammonia and oil production on protein and carbohydrate substrates.Applied microbiology and biotechnology · 2015Article
- Recent advances in DNA assembly technologies.FEMS yeast research · 2015Review
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4 authors at 2 institutions in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
Balancing the flux of a heterologous metabolic pathway by tuning the expression and properties of the pathway enzymes is difficult, but it is critical to realizing the full potential of microbial biotechnology. One prominent example is the metabolic engineering of a Saccharomyces cerevisiae strain harboring a heterologous xylose-utilizing pathway for cellulosic-biofuel production, which remains a challenge even after decades of research. Here, we developed a combinatorial pathway-engineering approach to rapidly create a highly efficient xylose-utilizing pathway for ethanol production by exploring various combinations of enzyme homologues with different properties. A library of more than 8,000 xylose utilization pathways was generated using DNA assembler, followed by multitiered screening, which led to the identification of a number of strain-specific combinations of the enzymes for efficient conversion of xylose to ethanol. The balancing of metabolic flux through the xylose utilization pathway was demonstrated by a complete reversal of the major product from xylitol to ethanol with a similar yield and total by-product formation as low as 0.06 g/g xylose without compromising cell growth. The results also suggested that an optimal enzyme combination depends on not only the genotype/phenotype of the host strain, but also the sugar composition of the fermentation medium. This combinatorial approach should be applicable to any heterologous pathway and will be instrumental in the optimization of industrial production of value-added products.
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