Dataset Information

Complete Genome Sequence of Megaplasmid-Bearing Streptococcus salivarius Strain LAB813, Isolated from the Dental Plaque of a Caries-Free Child.

ABSTRACT: Streptococcus salivarius strain LAB813 was isolated from the dental plaque biofilm of a caries-free child with healthy oral tissues. We report here the complete genome sequence of S. salivarius strain LAB813. This genome consists of a chromosome of 2.2 Mb and a megaplasmid, pSAL813, of 183 kb.

SUBMITTER: Gong SG

PROVIDER: S-EPMC6787328 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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Complete Genome Sequence of Megaplasmid-Bearing Streptococcus salivarius Strain LAB813, Isolated from the Dental Plaque of a Caries-Free Child.

Gong Siew-Ging SG Chan Yuki Y Lévesque Céline M CM

Microbiology resource announcements 20191010 41

<i>Streptococcus salivarius</i> strain LAB813 was isolated from the dental plaque biofilm of a caries-free child with healthy oral tissues. We report here the complete genome sequence of <i>S. salivarius</i> strain LAB813. This genome consists of a chromosome of 2.2 Mb and a megaplasmid, pSAL813, of 183 kb. ...[more]

PMID: 31601671

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Project description:The hydrolysis of urea by urease enzyme of oral bacteria is believed to have a major impact on oral microbial ecology and to be intimately involved in oral health and diseases. To begin to understand the biochemistry and genetics of oral ureolysis, a study of the urease of Streptococcus salivarius, a highly ureolytic organism which is present in large numbers on the soft tissues of the oral cavity, has been initiated. By using as a probe a 0.6-kpb internal fragment of the S. salivarius 57.I ureC gene, two clones from subgenomic libraries of S. salivarius 57.I in an Escherichia coli plasmid vector were identified. Nucleotide sequence analysis revealed the presence of one partial and six complete open reading frames which were most homologous to ureIAB-CEFGD of other ureolytic bacteria. Plasmid clones were generated to construct a complete gene cluster and used to transform E. coli and Streptococcus gordonii DL1, a nonureolytic, dental plaque microorganism. The recombinant organisms expressed high levels of urease activity when the growth medium was supplemented with NiCl2. The urease enzyme was purified from E. coli, and its biochemical properties were compared with those of the urease produced by S. salivarius and those of the urease produced by S. gordonii carrying the plasmid-borne ure genes. In all cases, the enzyme had a Km of 3.5 to 4.1 mM, a pH optimum near 7.0, and a temperature optimum near 60 degrees C. S. gordonii carrying the urease genes was then demonstrated to have a significant capacity to temper glycolytic acidification in vitro in the presence of concentrations of urea commonly found in the oral cavity. The ability to genetically engineer plaque bacteria that can modulate environmental pH through ureolysis will open the way to using recombinant ureolytic organisms to test hypotheses regarding the role of oral ureolysis in dental caries, calculus formation, and periodontal diseases. Such recombinant organisms may eventually prove useful for controlling dental caries by replacement therapy.

Project description:Due to the complex microecology and microenvironment of dental plaque, novel caries prevention strategies require modulating the microbial communities ecologically and reducing the cariogenic properties effectively. Antimicrobial peptide GH12 reduced the lactic acid production and exopolysaccharide (EPS) synthesis of a Streptococcus mutans biofilm and a three-species biofilm in vitro in previous studies. However, the anticaries effects and microecological effects of GH12 remained to be investigated in a complex biofilm model in vitro and an animal caries model in vivo In the present study, GH12 at 64 mg/liter showed the most effective inhibition of lactic acid production, EPS synthesis, pH decline, and biofilm integrity of human dental plaque-derived multispecies biofilms in vitro, and GH12 at 64 mg/liter was therefore chosen for use in subsequent in vitro and in vivo assays. When treated with 64-mg/liter GH12, the dental plaque-derived multispecies biofilms sampled from healthy volunteers maintained its microbial diversity and showed a microbial community structure similar to that of the control group. In the rat caries model with a caries-promoting diet, 64-mg/liter GH12 regulated the microbiota of dental plaque, in which the abundance of caries-associated bacteria was decreased and the abundance of commensal bacteria was increased. In addition, 64-mg/liter GH12 significantly reduced the caries scores of sulcal and smooth surface caries in all locations. In conclusion, GH12 inhibited the cariogenic properties of dental plaque without perturbing the dental plaque microbiota of healthy individuals and GH12 regulated the dysbiotic microbial ecology and arrested caries development under cariogenic conditions.IMPORTANCE The anticaries effects and microecological regulation effects of the antimicrobial peptide GH12 were evaluated systematically in vitro and in vivo GH12 inhibited the cariogenic virulence of dental plaque without overintervening in the microbial ecology of healthy individuals in vitro GH12 regulated the microbial ecology of dental plaque to a certain extent in vivo under cariogenic conditions, increased the proportion of commensal bacteria, and decreased the abundance of caries-associated bacteria. GH12 significantly suppressed the incidence and severity of dental caries in vivo This study thus describes an alternative antimicrobial therapy for dental caries.

Dataset Information

Complete Genome Sequence of Megaplasmid-Bearing Streptococcus salivarius Strain LAB813, Isolated from the Dental Plaque of a Caries-Free Child.

Publications

Complete Genome Sequence of Megaplasmid-Bearing Streptococcus salivarius Strain LAB813, Isolated from the Dental Plaque of a Caries-Free Child.

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