Isolation of Thermophilic Bacteria Geobacillus subterraneus From Mount Tangkuban Perahu and the Novelty as a Candidate for Streptococcus mutans Anti-Biofilm

Abstract

Thermophilic bacteria living in extreme areas with high temperatures are capable of producing secondary metabolites, such as antimicrobial peptides (AMPs). AMPs are stable at high temperatures and show good antibacterial activity. Therefore, this study aimed to identify thermophilic bacteria from the crater of Mount Tangkuban Perahu around West Java and assess antibacterial effectiveness of AMPs against Streptococcus mutans, which contribute to oral biofilm formation. The isolate obtained was identified using 16S ribosomal ribonucleic acid (rRNA) gene sequencing, and the supernatant of the isolate was tested against S. mutans American Type Culture Collection (ATCC) 25175 using the disc assay method. To determine AMPs-coding genes, its genome was uploaded to antibiotic and secondary metabolite analysis shell (antiSMASH) 5.0.0 platform and biofilm inhibition was tested using the microtiter plate technique (with a 96-well bottom). Subsequently, the results were assessed using a microplate reader operating at 595 nm wavelength. The isolate was identified as Geobacillus subterraneus, with antibacterial activity against S. mutans, and produced an inhibition zone of 8.40 mm at an optimum pH of 8. The output of AMPs-coding gene showed that AMPs of the isolate were a member of the lanthipeptide class I, or bacteriocin-I group. AMPs of G. subterraneus suppressed the growth of S. mutans biofilm at a supernatant concentration of 5%, with the lowest optical density (OD) value of 0.061 and the highest percentage of biofilm growth inhibition at 28.24%. Based on the results, G. subterraneus derived from the crater of Mount Tangkuban Perahu showed potent antibacterial properties against S. mutans, making it a promising novel S. mutans anti-biofilm candidate.