Genomic Analysis Of A.pasteulqanus CGMCC 15730 From Shanxi Aged Vinegar And Interactions With Other Excellent Strains

Acetobacter pasteurianus CGMCC 15730 is isolated from Shanxi aged vinegar,because of its strong abilities to produce and tolerate high concentrations of acetic acid.In this study,the genome of A.pasteurianus CGMCC 15730 was sequenced based on the Nanopore sequencing technology platform,which enriched the genomics of A.pasteurianus.At the same time,the microbial culture method was used to study the interaction between A.pasteurianus CGMCC 15730 and other excellent acetic acid bacteria,lactic acid bacteria,and bacillus,providing a theoretical basis for optimizing the fermentation process of Shanxi aged vinegar.The results are as follows:(1)The genome of A.pasteurianus CGMCC 15730 was sequenced using the Nanopore sequencing technology platform,and the genome was assembled,gene predicted,and functionally annotated with the sequencing data combined with bioinformatics analysis.The genome sequencing results showed that the genome of A.pasteurianus CGMCC 15730 was composed of a circular chromosome and two plasmids,the sizes were 2,760,994 bp,201,873 bp,and 246,670 bp,and the GC contents were 52.88%,51.75%,and 53.69%,respectively.In the chromosome,the average length of the gene was 973 bp,and a total of 2,549 CDS were predicted.In addition,the chromosome contains 15 complete r RNA gene manipulation units,56 t RNAs,1 CRISPR sequence,3 pseudogenes,and 2 prophages.On plasmid 1,the average length of the gene is 637 bp,containing 255 CDS and 6 pseudogenes.On plasmid 2,the average length of the gene is 625 bp,with 317 CDS and 10 pseudogenes.Based on the gene ontology database to annotate the protein-coding genes,the results show that 45.44% of the protein-coding genes of A.pasteurianus CGMCC15730 play an important role in the biological process in which they participate.The COG annotation was performed on protein-coding genes based on the Clusters of Orthologous Groups database.The results showed that the protein-coding genes of A.pasteurianus CGMCC15730 were mainly concentrated in gene replication,recomination and repair,amino acid transport and metabolism,energy production and conversion,etc.The KEGG annotation was performed on the protein-coding genes.The results showed that the gene of A.pasteurianus CGMCC15730 mainly play important roles in biosynthesis of amino acids,carbon metabolism and ABC transporters.Colinearity analysis based on whole-gene sequencing results showed that A.pasteurianus 386 B,A.pasteurianus Ab3,and 8 strains of A.pasteurianus IFO had highly similar genome sequences and the closest genetic relationship.(2)The interactions between A.pasteurianus CGMCC 15730,A.pasteurianus A33,Acetobacter rancens A8-7,and Acetobacter hansenii J2-5 in a pure and co-culture system were compared.It was found that co-cultivation of A.pasteurianus CGMCC 15730 and A.pasteurianus A33 promoted the growth of A.pasteurianus A33,increased the utilization of reducing sugars,total acid and acetoin content,and also increased Content of flavor substances such as volatile flavour compounds and organic acid.Under different temperature conditions,the interaction between A.pasteurianus CGMCC 15730 and Lactobacillus plantarum L7,L.plantarum L19,Lactobacillus lactis L729,and Pediococcus pentosaceus L2422 was studied.The results showed that when co-cultivated with A.pasteurianus CGMCC 15730 and L.plantarum L7 at 30°C,the total acid content increased by 74.49% compared with pure culture,and was significantly higher than the total acid content at 37°C.Organic acids such as acetic acid and acetic acid,and volatile flavour components such as acetoin and 2,3,5,6-tetramethylpyrazine increased significantly.And when the ratio of the two is 1:0.5,the growth of A.pasteurianus CGMCC 15730 is promoted,and the total acid(4.12 g / 100g),nonvolatile acid(1.63 g / 100),and acetoin(2.50 mg / g)content increased significantly,the total amount of organic acids(3.9095 g / 100g)increased,the content of volatile flavour compounds increased and the types increased.The interactions between A.pasteurianus CGMCC 15730 and Bacillus amyloliquefaciens B2030,Bacillus subtilis B803,B.amyloliquefaciens B1539 were studied.The results showed that at 37°C,the A.pasteurianus CGMCC 15730 and B.amyloliquefaciens B2030 co-cultivation is beneficial for acid production,ester production and acetoin production.The content of organic acids such as oxalic acid(0.3656g/100g),lactic acid(0.4520g/100g)and acetic acid(0.9525g/100g)is increased.The content of volatile flavour components such as 2,3-butanedione,2,3,5,6-tetramethylpyrazine and guaiacol increased.And it was better than co-culture of both at 30?.