Isolation and identification of microorganisms in pit mud and primary analysis of caproic acid bacteria metabolites
The strains were isolated from strong-flavor(Nongxiangxing)Baijiu pit mud by a variety of medium enrichment methods and anaerobic iso-lation techniques,their basic taxonomic information was determined by morphological observation,16S rRNA gene sequencing,and phylogenetic analysis,and the metabolites of caproic acid bacteria of Oscillospiraceae were analyzed by GC-MS and HPLC.The results showed that a total of 63 strains of bacteria were isolated.Based on the 16S rRNA gene sequences,all strains could be identified to genus and belonged to 2 class,3 order,8 fami-lies and 15 genus,including Clostridium,Lactiplantibacillus,Staphylococcus,Sporolactobacillus,Haloimpatiens,Paraclostridium,Rummeliibacillus,Caproiciproducens,etc.Among them,there were 9 strains showed low sequence similarity(96.17％-97.70％)to the proximate bacteria,which might be potential new species.Both of 3 caproic acid bacteria of Oscillospiraceae was capable of producing acetic acid,butyric acid,caproic acid by fermen-tation using RCM medium,and the caproic acid production of Caproicibacterium sp.20-5D,Caproiciproducens sp.CPB-2 and Caproiciproducens sp.YR-4 was 3.84 g/L,1.03 g/L and 0.35 g/L,respectively,with strong ability of producing mixed acids.The functional bacteria isolated from the pit mud of strong-flavor Baijiu had the great significance for enriching the numbers of culturable microbes in pit mud and the application and development of functional microorganisms.
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