Construction method and application of saccharomyces cerevisiae engineering bacteria knocked in COQ2 gene based on CRISPR-Cas9 system
By knocking in COQ2 gene based on the CRISPR-Cas9 system in Saccharomyces cerevisiae, the problem of difficulty in accumulating 5-DMQ3B during the fermentation of angelica is solved, and efficient and rapid 5-DMQ3B production is achieved, providing the possibility of application of this compound.
CN119709820APending Publication Date: 2025-03-28ZHEJIANG UNIV OF TECH
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Patent Information
- Application Number
- CN202411809313.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-10
- Publication Date
- 2025-03-28
AI Technical Summary
Technical Problem
The prior art is difficult to efficiently produce 5-DMQ3B because it is an intermediate metabolite and is difficult to accumulate during the fermentation of angelica.
Method used
Through the construction method of Saccharomyces cerevisiae engineered bacteria knocking in COQ2 gene based on the CRISPR-Cas9 system, efficient fermentation and production of 5-DMQ3B is achieved. The method includes steps such as COQ2 plasmid construction, gRNA plasmid construction, Cas9 plasmid transformation and co-transformation.
Benefits of technology
Through this method, 5-DMQ3B can be produced rapidly and in large quantities, providing the possibility of application of the compound, reducing production costs, and increasing the accumulation of 4-AAQB biosynthesis intermediates.
✦ Generated by Eureka AI based on patent content.
Abstract
The invention relates to a construction method and application of saccharomyces cerevisiae engineering bacteria knocked into COQ2 gene based on a CRISPR-Cas9 system. The construction method specifically comprises the following steps: 1) constructing COQ2 plasmid; the construction method comprises the following steps of (1) preparation of a recombinant expression system, (2) construction of gRNA plasmids, (3) preparation of wxc5 saccharomyces cerevisiae containing cas9 plasmids, (4) construction of Donor DNA, and (5) co-transformation.According to the construction method, a saccharomyces cerevisiae recombinant expression system is established, 5-DMQ3B is generated in saccharomyces cerevisiae engineering bacteria, a new method is provided for improving production of 5-DMQ3B, and the saccharomyces cerevisiae has the advantages of being low in fermentation production cost, convenient in raw material preparation, wide in industrial application prospect and the like.
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