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Production of pyruvate in Saccharomyces cerevisiae through adaptive evolution and rational cofactor metabolic engineering

Paper ID Volume ID Publish Year Pages File Format Full-Text
3513 173 2012 6 PDF Available
Title
Production of pyruvate in Saccharomyces cerevisiae through adaptive evolution and rational cofactor metabolic engineering
Abstract

Pyruvate-decarboxylase (Pdc)-negative Saccharomyces cerevisiae has been proven as a suitable metabolic engineering platform to produce organic acids. S. cerevisiae BY5419 Pdc− strain cannot grow in batch cultures on synthetic medium with glucose as the sole carbon source, yet grows well on synthetic medium with ethanol or acetate. In this study, by combining adaptive evolution and cofactor engineering, we obtained a series of engineered yeasts that can produce pyruvate using glucose as sole carbon source. Differential expression of noxE, encoding a water-forming NADH oxidase from Lactococcus lactis, and udhA, encoding a soluble pyridine nucleotide transhydrogenase from Escherichia coli, was investigated. Of all the constructed recombinant strains, G2U1-A0 was able to produce 75.1 g l−1 pyruvate, increased 21% compared with the original strain A0. The production yield of this strain reached 0.63 g of pyruvate g of glucose−1. This study demonstrates that the fine regulation of intracellular NADH/NAD+ ratio is critical for cell metabolism and pyruvate production. Combining the adaptive evolution and fine regulation of intracellular NADH/NAD+ ratio provides a new strategy for improving the Pdc− strain engineering platform.

► Combining adaptive evolution and cofactor engineering obtained engineered yeasts. ► G2U1-A0 was able to produce 75.1 g l−1 pyruvate. ► Differential expression of noxE and udhA was investigated. ► Fine regulation of NADH/NAD+ ratio is critical to pyruvate production. ► A new strategy for improvement of the Pdc− strain engineering platform is provided.

Keywords
NADH/NAD+; Pyruvate; Saccharomyces cerevisiae; Metabolic engineering; Adaptive evolution; Cofactor metabolic engineering
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Production of pyruvate in Saccharomyces cerevisiae through adaptive evolution and rational cofactor metabolic engineering
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 67, 15 August 2012, Pages 126–131
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us