Genome sequence analisys of Lactobacilli strain isolated from fermented animal food

PABLO MORTERA; VÍCTOR S. BLANCATO; MARTÍN ESPARIZ; CHRISTIAN MAGNI

Simposio; V Simposio Internacional de Bacterias Lácticas (SIBAL); 2016

Lactobacilli are a common lactic acid bacteria (BAL) widely studied in regard to their role in the dairy technology and probiotic properties. New isolated strains from specific environments, allows us to identify physiological capacities of interest associated with novelmetabolic networks. A Lactobacilli strain, named PM07, was isolated from fermented cereal food using for livestock feeding by the Nutreza company, in Santa Fe. Part of our biotechnological study, comprises to find fibrinolytic activities (xylanase, cellulase, chitinase oramylase) and another catabolic pathways present in this strain associated to vegetal fermentation. Illumina technology was used for sequence and SeqMan NGen software, Advanced Pipmaker and Mauve Genome Alignment for genome sequences assembly.Phylogenetic genome analysis, performed with Gegenees, provided the strain PM07 present 84-86% similarity with 11 Lb. casei and 84-85% with 2 Lb. paracasei strains. The similarity with the model strain Lb. casei BL23 was 84%. Genome annotation by RAST. Genomesize was 2,995,822 bases in length with 46,4% GC content, containing 2975 coding sequences (CDS) and 71 RNAs (61 tRNAs) predicted. The Genome annotation, achieved by RAST covered 341 subsystems, including 1276 (43%) of which 63 CDS were labeled ashypothetical proteins. As most BAL, Lb. PM07 strain shows a wide sugar fermentation capability, represented by 425 total CDS related to carbohydrates metabolism. Specifically, a total of 10 genes encoding enzymes for maltose and maltodextrin utilization, and 26genes related to lactose and galactose uptake and utilization, 12 of them are in a cluster organization. Chitin and N-acetylglucosamine utilization were assigned to the presence of 14 genes. Likewise, a total of 11 genes were associated to N-acetylgalactosamine andgalactosamine metabolism. Instead, no genes related to cellulose metabolism were detected. Noteworthy, we found an 8 genes cluster encoding among others proteins, an aspartate aminotransferase family enzyme, a putative transport system permease protein,phosphopentomutase like enzyme, phosphotriesterase, and evolved beta-D-galactosidase. Most of these genes are only presents in few Lactobacilli strains (Lactobacillus casei LOCK919, Lb. paracasei ATCC 334 and 2 strains of Lb. rhamnosus). Moreover, we alsodetected 35 CDS corresponding to phage/prophage proteins machine. PHAST analysis tool revealed at least 4 prophage regions identified. Non plasmidic, virulence factors either human pathogenic sequences were found using Finder genomic analysis tools. Additionalbioinformatical and physiological works are needed to classify phylogenetically the Lactobacillus PM07 strain as casei or paracasei and evaluate the potential technological to be used in fermented feed