An escherichia coli engineering bacterium for producing batroxobin with controllable generation and a construction method and application thereof

By constructing a hokC toxin protein expression vector and combining it with the constitutive promoter trxA fragment, the problem of imprecise control of hirudin production in engineered Escherichia coli strains was solved. Active response and production regulation were achieved under conditions without inducers, making it applicable to a variety of hirudin-synthesizing strains and meeting the production needs of the biopharmaceutical field.

CN121826015BActive Publication Date: 2026-07-14INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
Filing Date
2026-03-11
Publication Date
2026-07-14

AI Technical Summary

Technical Problem

In existing technologies, the production control of engineered hirudin-producing Escherichia coli is not precise enough, making it difficult to actively respond without inducers. Furthermore, leaked engineered bacteria may survive and spread for a long time, affecting the safety and efficiency of industrial production.

Method used

The hokC toxin expression vector pET28a(+)-ptrxA-hokC was constructed and transformed into a hirudin-producing strain. The hirudin-producing Escherichia coli strain was constructed by regulating the hirudin production through the constitutive promoter trxA fragment. The hirudin production was finely regulated through multiple rounds of culture.

Benefits of technology

It achieves active response and precise control of hirudin production under inducer-free conditions, enhances the convenience and autonomy of regulation, adapts to industrial production needs, is applicable to a variety of hirudin-synthesizing strains, and has versatility.

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Abstract

The application discloses an E. coli engineering bacterium for producing controllable-generation hirudin and a construction method and application thereof, and relates to the technical field of biological medicines. hokC The construction method of the E. coli engineering bacterium for producing controllable-generation hirudin is to construct a recombinant expression vector pET28a(+)-ptrxA-hokC by using a protein gene and a constitutive promoter trxA with a sequence as shown in SEQ ID NO. 2, and then transform the recombinant expression vector into a hirudin synthesis strain E. coli Hirudin HV1-3 to obtain the E. coli engineering bacterium for producing controllable-generation hirudin. After multiple rounds of culture, the yield of hirudin of the engineering strain is reduced to 0 at the 10th round. The application is suitable for various types of hirudin and has the characteristics of reliable operation and high safety.
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