Pyruvate gene loop mediated saccharomyces cerevisiae ethanol metabolism redirection method

By constructing a pyruvate response gene circuit in Saccharomyces cerevisiae, dynamically regulating cell growth and product synthesis of Saccharomyces cerevisiae, the problem of restriction of ethanol synthesis under high sugar conditions is solved, the recovery of cell growth and the maximization of product synthesis flux is achieved, the fermentation cycle is shortened and the production efficiency is improved.

CN120442687APending Publication Date: 2025-08-08NANJING TECH UNIV
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Patent Information

Application Number
CN202510661420.2
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-05-22
Publication Date
2025-08-08

AI Technical Summary

Technical Problem

Saccharomyces cerevisiae preferentially fermented through glycolytic pathways under high sugar conditions, resulting in carbon flux flow to ethanol synthesis, limiting the yield and cell growth of non-ethanol biological products. The static gene knockout strategy leads to impaired cell growth and imbalance in cofactor reduction. The existing dynamic regulatory methods are insufficiently used in Saccharomyces cerevisiae.

Method used

The pyruvate response gene circuit is constructed in Saccharomyces cerevisiae, and the cell growth and product synthesis are dynamically regulated during the fermentation process through the bifunctional pyruvate gene circuit, and the gene expression is controlled using pyruvate activated and inhibitory promoters, combined with the glucose stage supply strategy, and the dynamic balance of cell growth, ethanol synthesis and product synthesis is achieved.

Benefits of technology

The dynamic recovery of Saccharomyces cerevisiae cell growth and the maximum product synthesis flux are achieved, the fermentation cycle is shortened, the product production intensity and efficiency are improved, and it is suitable for the fermentation and production of different metabolites.

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Abstract

The invention discloses a pyruvic acid gene loop mediated saccharomyces cerevisiae ethanol metabolism redirection method. Comprising the following steps: constructing a pyruvic acid response gene loop in saccharomyces cerevisiae host bacteria to obtain modified saccharomyces cerevisiae; the growth of saccharomyces cerevisiae cells, the synthesis of by-product ethanol and the synthesis of products are dynamically regulated and modified by controlling glucose feeding. A low-concentration glucose environment is maintained in the early stage of growth, so that cell growth and ethanol metabolism related genes are normally expressed, and cell growth is recovered; high-concentration glucose is supplemented in the later stage of fermentation, expression of cell growth related ethanol metabolism genes is inhibited, the ethanol synthesis flux is reduced, sufficient pyruvic acid precursors are provided for product synthesis, and the product synthesis flux is maximized. According to the method, the yield of the byproduct ethanol is reduced, the production efficiency of the product is improved, and meanwhile, the fermentation period is shortened.
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