A combination module and its application in improving stress resistance of nitrogen-fixing microorganisms

By constructing a Pc-algU-dosH combined module in nitrogen-fixing microorganisms, the problems of long time consumption, high cost and poor versatility in the existing technology are solved, which significantly improves the stress resistance of nitrogen-fixing microorganisms under high temperature and drought conditions and enhances nitrogen fixation efficiency.

CN117165619BActive Publication Date: 2026-07-24THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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Patent Information

Application Number
CN202311049843.6
Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Filing Date
2023-08-18
Publication Date
2026-07-24
Estimated Expiration
2043-08-18

AI Technical Summary

Technical Problem

Existing technologies for constructing the stress resistance of nitrogen-fixing microorganisms suffer from problems such as long time consumption, high cost, poor versatility, and low level of intelligence. Furthermore, existing synthetic biology methods have poor tolerance to adverse conditions or are only tolerant to a single condition.

Method used

Synthetic biology techniques were used to construct a combinatorial module, including the constitutive promoter element Pc, the RNA polymerase σ factor AlgU from Pseudomonas, and the molecular chaperone protein DosH from Anomococcus. This module was then introduced into a recombinant expression vector using seamless cloning technology to form an expression cassette for stress tolerance proteins. The vector was then transferred into nitrogen-fixing microorganisms to construct recombinant strains.

Benefits of technology

It significantly improved the stress resistance of nitrogen-fixing microorganisms under high temperature and drought stress, enhanced nitrogenase activity and survival ability, and increased nitrogen fixation efficiency in the plant rhizosphere.

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Abstract

The application provides a combination module and application thereof in improving stress resistance of nitrogen-fixing microorganisms. The combination module comprises, in sequence from upstream to downstream, a constitutive promoter element Pc, a RNA polymerase sigma factor from Pseudomonas AlgU and a molecular chaperone protein from Deinococcus DosH . The application also provides application of the combination module in improving stress resistance of nitrogen-fixing microorganisms. The application obtains the combination module Pc −algU−dosH −algU−dosH , introduces the combination module into an expression vector to construct a recombinant expression vector, and transfers the recombinant vector into nitrogen-fixing bacteria to obtain a nitrogen-fixing recombinant engineering strain. Compared with a single stress resistance module, the combination module can significantly improve the stress resistance of a chassis nitrogen-fixing microorganism under high-temperature and drought stress conditions, and significantly improve the nitrogenase activity and survival ability of the chassis microorganism.
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