High efficient screening method for the uncultured microorganisms in Daqu, distiller's grains and pit mud based on the concept of in-situ culture

By using in-situ culture technology and a 96-well plate device, combined with a specific simulated matrix, the problem of difficult-to-culture microorganisms in the Baijiu brewing system that are difficult to screen using the traditional plate method has been solved. This has enabled the isolation and purification of more strains, expanded the types of culturable microorganisms, and promoted the development of microbial resources in the Baijiu brewing system.

CN122357282APending Publication Date: 2026-07-10GUIZHOU UNIV +1
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
GUIZHOU UNIV
Filing Date
2026-04-01
Publication Date
2026-07-10

AI Technical Summary

Technical Problem

The existing traditional plate culture method is difficult to effectively screen and culture a variety of difficult-to-culture microbial resources in the Baijiu brewing system, resulting in a large number of potential functional strains remaining undeveloped.

Method used

Using in-situ culture technology, a unique microbial in-situ culture device was constructed by combining a 0.2μm nylon filter and a 96-well plate. Specific in-situ environment simulation substrates of Daqu (fermentation starter), distiller's grains, and cellar mud were designed to simulate the baijiu brewing environment. Through gradient dilution and in-situ culture enrichment, the strains were then isolated and purified.

Benefits of technology

It significantly improved the attachment and growth efficiency of difficult-to-cultivate microorganisms, expanded the range of culturable microorganisms, and successfully isolated and screened a variety of strains that were difficult to isolate using traditional plate methods, such as Talamoyces aculeatus and Scedosporium boydii, providing technical assistance for the development of microbial 'dark matter' resources in the Baijiu brewing system.

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Abstract

The application designs a microbial separation and screening scheme, through 0.2 mu m food-grade nylon filter screen and special in-situ culture device of 96-hole plate, and is embedded in three in-situ environment simulation matrixes (Daqu, vinasse, pit mud) to ensure good growth and metabolism adaptability of difficult-to-culture strains in the enrichment process, reduce the interference of external adverse factors, effectively realize the directional enrichment of difficult-to-culture microbial groups in traditional Daqu, vinasse, pit mud and other liquor fermentation matrixes, and expand the population range of cultivable microorganisms in the liquor brewing environment. Finally, through the actual application of the technical scheme, a plurality of difficult-to-culture strains have been successfully separated and screened from Daqu, vinasse, pit mud and other samples, and a method reference is provided for efficient development and application of more microbial 'dark matter' resources with good application potential in the traditional liquor brewing system in the future.
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