Dataset Information

Whole-Genome Sequence of Lactobacillus sakei LT-13 Isolated from Moto Starter of Sake.

ABSTRACT: Lactobacillus sakei strain LT-13 is a lactic acid bacterium isolated from moto starter of Japanese sake. This genome analysis revealed that the genome is composed of a circular chromosome and one plasmid, which contain 1,938 and 8 putative protein-coding genes, respectively.

SUBMITTER: Kato S

PROVIDER: S-EPMC5543631 | biostudies-literature | 2017 Aug

REPOSITORIES: biostudies-literature

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Whole-Genome Sequence of <i>Lactobacillus sakei</i> LT-13 Isolated from Moto Starter of Sake.

Kato Shiro S Oikawa Tadao T

Genome announcements 20170803 31

<i>Lactobacillus sakei</i> strain LT-13 is a lactic acid bacterium isolated from moto starter of Japanese sake. This genome analysis revealed that the genome is composed of a circular chromosome and one plasmid, which contain 1,938 and 8 putative protein-coding genes, respectively. ...[more]

PMID: 28774969

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Project description:The Lactobacillus sakei strain LK-145 isolated from Moto, a starter of sake, produces potentially large amounts of three D-amino acids, D-Ala, D-Glu, and D-Asp, in a medium containing amylase-digested rice as a carbon source. The comparison of metabolic pathways deduced from the complete genome sequence of strain LK-145 to the type culture strain of Lactobacillus sakei strain LT-13 showed that the L- and D-amino acid metabolic pathways are similar between the two strains. However, a marked difference was observed in the putative cysteine/methionine metabolic pathways of strain LK-145 and LT-13. The cystathionine ?-lyase homolog gene malY was annotated only in the genome of strain LT-13. Cystathionine ?-lyase is an important enzyme in the cysteine/methionine metabolic pathway that catalyzes the conversion of L-cystathionine into L-homocysteine. In addition to malY, most genome-sequenced strains of L. sakei including LT-13 lacked the homologous genes encoding other putative enzymes in this pathway. Accordingly, the cysteine/methionine metabolic pathway likely does not function well in almost all strains of L. sakei. We succeeded in cloning and expressing the malY gene from strain LT-13 (Ls-malY) in the cells of Escherichia coli BL21 (DE3) and characterized the enzymological properties of Ls-MalY. Spectral analysis of purified Ls-MalY showed that Ls-MalY contained a pyridoxal 5'-phosphate (PLP) as a cofactor, and this observation agreed well with the prediction based on its primary structure. Ls-MalY showed amino acid racemase activity and cystathionine ?-lyase activity. Ls-MalY showed amino acid racemase activities in various amino acids, such as Ala, Arg, Asn, Glu, Gln, His, Leu, Lys, Met, Ser, Thr, Trp, and Val. Mutational analysis revealed that the ?-amino group of Lys233 in the primary structure of Ls-MalY likely bound to PLP, and Lys233 was an essential residue for Ls-MalY to catalyze both the amino acid racemase and ?-lyase reactions. In addition, Tyr123 was a catalytic residue in the amino acid racemase reaction but strongly affected ?-lyase activity. These results showed that Ls-MalY is a novel bifunctional amino acid racemase with multiple substrate specificity; both the amino acid racemase and ?-lyase reactions of Ls-MalY were catalyzed at the same active site.

Project description:Kimoto-style seed mash is a traditional preparation method for sake that takes advantage of spontaneous lactic acid fermentation before the growth of yeast. Lactic acid helps decrease the pH in seed mash and control the growth of unfavorable microorganisms. In this study, we carried out a comprehensive analysis of the change in the bacterial community and chemical composition during the lactic acid fermentation stage in kimoto-style seed mash preparation. The bacterial transitions were diverse at five sake breweries, but they exhibited three patterns. Lactobacillus sakei was the dominant species in the later stage of lactic acid fermentation in all sake breweries. This species was found to be the most important bacterium for the accumulation of lactic acid, because its average production rate of lactic acid in seed mash reached 4.44 × 10-11 mg cell-1 h-1, which is 10 times higher than those of other species. As a result of specific growth rate analysis, it was revealed that the growth rate of L. sakei was influenced by the strain, pH, and temperature. The effects of pH and temperature were explained by the square root model, and the result indicates that the strains isolated in this study were incapable of growth below pH 3.9. The growth curve predicted using the growth model fit the actual cell density in two out of five sake breweries; however, our model did not work well for the remaining three sake breweries, and we presume that the error was caused by the strain or an unknown factor.IMPORTANCE It is important to produce lactic acid in kimoto-style seed mash; however, the bacterial transition is different depending on the sake brewery. The reason why there are diverse bacterial transitions during kimoto-style seed mash preparation for each sake brewery is unclear so far, and it causes difficulty in starting kimoto-style seed mash. Our findings indicate that the changes in pH caused by lactic acid bacteria grown prior to L. sakei in seed mash influence the growth of L. sakei and are related to the diversity of the bacterial transition. This study uses comprehensive analytical methods to reveal that there is a diversity of bacterial transition and chemical compositions in kimoto-style seed mash depending on the sake brewery and to explain the differences in bacterial transition depending on the characteristics of L. sakei.

Dataset Information

Whole-Genome Sequence of Lactobacillus sakei LT-13 Isolated from Moto Starter of Sake.

Publications

Whole-Genome Sequence of <i>Lactobacillus sakei</i> LT-13 Isolated from Moto Starter of Sake.

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