A mutant of expansin protein from rumen anaerobic fungus and its coding gene and application

By performing site-directed mutagenesis on the expansin protein derived from the rumen anaerobic fungus *Eriocaulon simonii* LGM-ZA9, the problems of low expression level and poor stability were solved, achieving efficient degradation of lignocellulose and reduction of methane emissions, thus improving the utilization efficiency and environmental friendliness of agricultural waste.

CN122036895BActive Publication Date: 2026-06-26NANJING AGRICULTURAL UNIVERSITY
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Patent Information

Application Number
CN202610499528.0
Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Filing Date
2026-04-16
Publication Date
2026-06-26
Estimated Expiration
2046-04-16

AI Technical Summary

Technical Problem

Existing commercial expansion hormones are mainly derived from aerobic microorganisms, which have limited stability and activity in anaerobic environments such as the rumen. Furthermore, the expansion hormone gene PFSWO2_1 derived from the anaerobic fungus Neocallimastix sp. has low expression levels in heterologous expression systems, which limits its industrial application. At the same time, methane emissions from the rumen of ruminants lead to the greenhouse effect and feed energy waste.

Method used

By performing site-directed mutagenesis on wild-type expansin protein PFSWO2_1 derived from *Echinococcus neoformans* LGM-ZA9, mutants PFSWO2_1-N1, PFSWO2_1-N2, and PFSWO2_1-N3 with higher expression levels, better thermal stability, and stronger synergistic effect with cellulase were obtained. These mutants can be used to prepare enzyme preparations that degrade lignocellulose and feed additives that reduce rumen methane emissions in ruminants.

Benefits of technology

The mutant significantly improved the enzymatic hydrolysis efficiency and feed utilization rate of lignocellulose, reduced methane emission intensity by 15-20%, improved the utilization efficiency of agricultural waste, and reduced greenhouse gas emissions by regulating the rumen fermentation pathway.

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Abstract

The application discloses a mutant of expansin protein from rumen anaerobic fungi, a coding gene and application thereof, and belongs to the technical field of biological genetic engineering. The wild-type expansin protein PFSWO2_1 is subjected to site-directed mutation, and mutants with higher expression, better thermal stability and stronger synergistic effect with cellulase, namely the mutants PFSWO2_1-N1, PFSWO2_1-N2 and PFSWO2_1-N3, are obtained. The mutants are successfully expressed in Pichia pastoris, the hydrogen bond network between cellulose molecules is destroyed, the mutants can significantly enhance the enzymolysis efficiency of cellulase on lignocellulose, the mutants can not only improve the utilization rate of agricultural wastes such as straws in feed, but also can inhibit the generation of methane by regulating the fermentation parameters and microbial community structure in the rumen in vitro fermentation process, and the mutants have important application value in reducing the greenhouse gas emission of the animal husbandry.
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