Escherichia coli alkali-resistant target spot and application thereof

By screening 43 novel gene target combinations in Escherichia coli through whole-genome screening, an alkali-tolerant engineered strain was constructed, which solved the problem of cell damage caused by alkaline products in the process of diamine synthesis in Escherichia coli and significantly improved alkali tolerance and production capacity.

CN121362766APending Publication Date: 2026-01-20NANJING TECH UNIV
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Patent Information

Application Number
CN202511789089.9
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-12-01
Publication Date
2026-01-20

AI Technical Summary

Technical Problem

In existing technologies, the pH value of diamine synthesis in Escherichia coli increases due to the alkalinity of the product, causing cell damage and limiting the yield, conversion rate, and efficiency of diamine. Furthermore, traditional breeding methods are inefficient.

Method used

Through high-throughput screening of genome-wide overexpression libraries, 43 novel gene targets related to alkali tolerance in Escherichia coli were systematically identified, especially combinations of metN, prpE, fruK, nikC, mazG, and ybiT. Recombinant plasmids were constructed and expressed in Escherichia coli to form engineered strains with synergistic effects.

Benefits of technology

Under extreme alkaline stress, the growth rate of Escherichia coli was upregulated by more than 3 times, significantly improving alkaline tolerance and providing a rich gene resource library, laying the foundation for improving the intensity and final yield of diamine production.

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Abstract

The invention discloses an Escherichia coli alkali-resistant target spot and application thereof, an inhibition model of pH on Escherichia coli E.coli AG1 is established by setting LB culture media with different pH values, and it is found that the inhibition degree of cell growth is positively correlated with the rise of pH. Under the condition that the pH value is equal to 10, the escherichia coli overexpression library ASKA is subjected to high-throughput screening, and 43 alkali-tolerant elements are obtained after multiple rounds of screening. The obtained gene is subjected to KEGG / GO enrichment analysis through an online gene function annotation database DAVID, an EcoCyc database and related literatures are looked up, an alkali tolerance mechanism is predicted, and it is found that the alkali tolerance mechanism is possibly related to biological membrane formation, intracellular pH steady state maintenance, intracellular ROS level reduction and the like. Through overexpression of the genes, the strain can adapt to an environment under alkali stress, the alkali tolerance of the strain is improved, and the strain has a good application prospect.
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