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      Antibodies generated against dextransucrase exhibit potential anticariostatic properties in Streptococcus mutans

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          Abstract

          Streptococcus mutans is a common principal causative agent of dental caries. In this communication, we describe that the antibodies raised against purified dextransucrase effectively inhibited the growth of S. mutans. The purified enzyme showed 58-fold enrichment, 17.5% yield and a specific activity of 3.96 units/mg protein. Purified IgG fraction of the antibody showed significant affinity with the antigenic protein. Immunotritation of the enzyme with dextransucrase antibody showed a gradual increase in inhibition of dextransucrase activity. The growth of S. mutans was also inhibited by 85% in the presence of 28 μg of IgG fraction of the antibody. Antibodies also impaired glucosyltransferase activity (72.8%) and biofilm formation by 92.6% in S. mutans. Western blot analysis revealed no cross reactivity with the various tissues of mice, rat, rabbit and humans. Dot blot analysis showed little reactivity with Lactobacillus acidophilus and Staphylococcus aureus and there was no reactivity with other bacterial strains like Enterococcus faecalis, Escherichia coli and Salmonella typhimurium. These findings suggest that antibody raised against dextransucrase exhibit inhibitory effects on the growth of S. mutans and biofilm formation with no reactivity with various mammalian tissues, thus it could be an effective anticariogenic agent.

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          Most cited references35

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          Functional genomics approach to identifying genes required for biofilm development by Streptococcus mutans.

          Streptococcus mutans, the primary etiological agent of human dental caries, is an obligate biofilm-forming bacterium. The goals of this study were to identify the gene(s) required for biofilm formation by this organism and to elucidate the role(s) that some of the known global regulators of gene expression play in controlling biofilm formation. In S. mutans UA159, the brpA gene (for biofilm regulatory protein) was found to encode a novel protein of 406 amino acid residues. A strain carrying an insertionally inactivated copy of brpA formed longer chains than did the parental strain, aggregated in liquid culture, and was unable to form biofilms as shown by an in vitro biofilm assay. A putative homologue of the enzyme responsible for synthesis of autoinducer II (AI-2) of the bacterial quorum-sensing system was also identified in S. mutans UA159, but insertional inactivation of the gene (luxS(Sm)) did not alter colony or cell morphology or diminish the capacity of S. mutans to form biofilms. We also examined the role of the homologue of the Bacillus subtilis catabolite control protein CcpA in S. mutans in biofilm formation, and the results showed that loss of CcpA resulted in about a 60% decrease in the ability to form biofilms on an abiotic surface. From these data, we conclude that CcpA and BrpA may regulate genes that are required for stable biofilm formation by S. mutans.
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            Fueling the caries process: carbohydrate metabolism and gene regulation by Streptococcus mutans

            The nature of the oral cavity and host behaviors has mandated that the oral microbiota evolve mechanisms for coping with environmental fluctuations, especially changes in the type and availability of carbohydrates. In the case of human dental caries, the presence of excess carbohydrates is often responsible for altering the local environment to be more favorable for species associated with the initiation and progression of disease, including Streptococcus mutans. Some of the earliest endeavors to understand how cariogenic species respond to environmental perturbations were carried out using chemostat cultivation, which provides fine control over culture conditions and bacterial behaviors. The development of genome-scale methodologies has allowed for the combination of sophisticated cultivation technologies with genome-level analysis to more thoroughly probe how bacterial pathogens respond to environmental stimuli. Recent investigations in S. mutans and other closely related streptococci have begun to reveal that carbohydrate metabolism can drastically impact pathogenic potential and highlight the important influence that nutrient acquisition has on the success of pathogens; inside and outside of the oral cavity. Collectively, research into pathogenic streptococci, which have evolved in close association with the human host, has begun to unveil the essential nature of careful orchestration of carbohydrate acquisition and catabolism to allow the organisms to persist and, when conditions allow, initiate or worsen disease.
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              Virulence factors of mutans streptococci: role of molecular genetics.

              Biochemical approaches were utilized initially to identify the virulence factors of the mutans streptococci (primarily Streptococcus mutans and S. sobrinu). Traditional mutant analysis of these organisms further suggested the important role of several of these factors in cariogenicity. However, because these mutations were not clearly defined, the utilization of cloned genes was necessary to verify their significance. The introduction of molecular genetic approaches for characterizing these factors has led not only to a clearer understanding of the role of these virulence factors in cariogenicity but has also suggested some novel approaches for reducing further the incidence of dental caries.
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                Author and article information

                Contributors
                rathershabeer@gmail.com
                sukeshcs@pu.ac.in
                akhtarmah@yahoo.com
                Journal
                Appl Microbiol Biotechnol
                Appl. Microbiol. Biotechnol
                Applied Microbiology and Biotechnology
                Springer Berlin Heidelberg (Berlin/Heidelberg )
                0175-7598
                1432-0614
                3 January 2020
                2020
                : 104
                : 4
                : 1761-1772
                Affiliations
                GRID grid.261674.0, ISNI 0000 0001 2174 5640, Department of Biochemistry, , Panjab University, ; Sector 14, Chandigarh, 160014 India
                Author information
                http://orcid.org/0000-0003-3614-3049
                Article
                10327
                10.1007/s00253-019-10327-x
                7223241
                31900558
                8585129c-564c-4bf1-b913-f97bd4d4e389
                © Springer-Verlag GmbH Germany, part of Springer Nature 2020

                This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.

                History
                : 18 September 2019
                : 11 December 2019
                : 18 December 2019
                Categories
                Applied Microbial and Cell Physiology
                Custom metadata
                © Springer-Verlag GmbH Germany, part of Springer Nature 2020

                Biotechnology
                dextransucrase,dental caries,streptococcus mutans,antibody,biofilm,anticariogenic agent
                Biotechnology
                dextransucrase, dental caries, streptococcus mutans, antibody, biofilm, anticariogenic agent

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