Abstract
Dental biofilms represent a serious oral health problem playing a key role in the development of caries and other oral diseases. In the present work, we cloned and expressed in E. coli two glucanases, Prevotella melaninogenica mutanase (PmGH87) and Capnocytophaga ochracea dextranase (CoGH66), and characterized them biochemically and biophysically. Their three-dimensional structures were elucidated and discussed. Furthermore, we tested the capacity of the enzymes to hydrolyze mutan and dextran to prevent formation of Streptococcus mutans biofilms, as well as to degrade pre- formed biofilms in low and abundant sugar conditions. The percentage of residual biofilm was calculated for each treatment group in relation to the control, as well as the degree of synergism. Our results suggest that both PmGH87 and CoGH66 are capable of inhibiting biofilm formation grown under limited or abundant sucrose conditions. Degradation of pre-formed biofilms experiments reveal a time-dependent effect for the treatment with each enzyme alone. In addition, a synergistic and dose-dependent effects of the combined enzymatic treatment with the enzymes were observed. For instance, the highest biomass degradation was 95.5% after 30 min treatment for the biofilm grown in low sucrose concentration, and 93.8% after 2 h treatment for the biofilm grown in sugar abundant condition. Strong synergistic effects were observed, with calculated degree of synergism of 5.54 and 3.18, respectively and their structural basis was discussed. Jointly, these data can pave the ground for the development of biomedical applications of the enzymes for controlling growth and promoting degradation of established oral biofilms.
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The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
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Funding
This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) via grants 2021/08780-1 and 2020/15092-1, and by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) via grants 306852/2021-7 and 440180/2022-8.
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All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by Anelyse Abreu Cortez, Mateus Xavier de Queiroz and Caio Cesar de Mello Capetti. The first draft of the manuscript was written by Anelyse Abreu Cortez and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Cortez, A.A., de Queiroz, M.X., de Oliveira Arnoldi Pellegrini, V. et al. Recombinant Prevotella melaninogenica α-1,3 glucanase and Capnocytophaga ochracea α-1,6 glucanase as enzymatic tools for in vitro degradation of S. mutans biofilms. World J Microbiol Biotechnol 39, 357 (2023). https://doi.org/10.1007/s11274-023-03804-z
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DOI: https://doi.org/10.1007/s11274-023-03804-z