Culturing The Baijiu Microorganism From Fermentation Vinasse And Pit Mud With Culturomics And Amplicon-based Metagenomic Approaches

Baijiu is a kind of Chinese traditional fermented liquor.The fermentation components possess plentiful microbial diversity.Based on amplicon-based metagenomics and culturomics,we investigated the microbial community structure and diversity of baijiu vinasse and pit mud.Finally,457 bacterial monoclones and 53 fungal monoclones were obtained.Metagenomic analysis showed that bacteria in vinasse belonged to 21 phyla,39 classes,84 orders,182 families,384 genera and 528 species.Among these,Lactobacillus exhibited the highest relative abundance.Bacteria in pit mud belonged to 9 phyla,23 classes,49 orders,99 families,201 genera and 313 species,most of which were anaerobic bacteria.Hydrogenispora possesseed the highest relative abundance.Fungi in vinasse belonged to 4phyla,16 classes,38 orders,70 families,107 genera and 164 species.Among these,Aspergillus exhibited the highest relative abundance.Fungi in pit mud belonged to 7 phyla,20 classes,37 orders,68 families,100 genera and 139 species.Pseudeurotium was the highest relative abundance.Based on the amplicon-based metagenomics analysis,culturomics was designed for isolating the bacteria and fungi in vinasse and pit mud.Totally 21 conditions were set for isolating vinasse's bacteria.And 242 strains were isolated,which belonged to 4 phyla,6classes,12 orders,21 families,32 genera and 84 species,which increased to 275% compared with previously reported.In addition,62 strains were classified as potential new species,belonged to 14 genera.Three new species were verified according to genome sequence.Tweenty-seven conditions were set for isolating pit mud's bacteria.Finally,215 strains bacteria were isolated,which belonged to 4 phyla,7 classes,13 orders,27 families,40 genera and 66 species,which increased to 252% compared with previously reports.In addition,48 strains were classified as potential new species,belonged to 11 genera.10 new species were verified according to genome sequence.Tweenty-one conditions were set for isolating the fungi in both vinasse and pit mud.Totally 33 fungi were obtained from vinasse,belonging to 5 genera.20 fungi were obtained from pit mud,belonging to 6 genera.This study analyzed the microbial community structure in baijiu vinasse and pit mud,and predicted their functions based on metagenomics analysis.Culturomics expanded the cultivable microbial species in baijiu vinasse and pit mud,and detected the genus with low abundance.Our study provides basis for cultivation and application of brewing microbes.