Secondary Metabolites of Halobacillus sp.: Antimicrobial and Antioxidant Activity, Biological Compatibility, and Gas Chromatography-Mass Spectrometry (GC-MS) Analysis.

ABSTRACT

Background The rise of infectious diseases and the emergence of resistant pathogens pose significant challenges to human health. In response to this global threat, researchers are exploring novel sources of bioactive compounds for effective antimicrobial therapies. One avenue of investigation is the study of halophilic bacteria and their secondary metabolites. These bacteria thrive under extreme conditions and produce valuable bioactive metabolites, which have the potential for therapeutic applications. Methods In this study, the potent bacterial cultures obtained from the Payanur salt pan, Tamil Nadu, were analyzed for the antimicrobial activity of their metabolites. The secondary metabolites were obtained from the halophilic bacteria by culturing the bacteria in 8% NaCl. The resultant secondary metabolites produced were extracted using ethyl acetate and their antimicrobial property was studied using the well diffusion method. The minimum inhibitory concentration (MIC) of these metabolites against five clinical pathogens, namely, Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, Pseudomonas aeruginosa, and Candida albicans was determined. Their antioxidant property was studied using the DPPH (2,2-diphenyl-1-picrylhydrazyl) method and biological compatibility was determined by hemolytic assay of the secondary metabolites. Results The potent halophilic bacteria isolated from salt pan bacteria were phenotypically and genotypically identified as Halobacillus sp. The secondary metabolites extracted from these bacteria yielded 110 mg of crude metabolites. The antimicrobial activity of crude metabolites shows a moderate zone of inhibition of 14 mm for P. aeruginosa, 13 mm for E. coli and C. albicans, and 11 mm for S. aureus. The minimum inhibitory concentration was 128 µg/mL for E. coli, P. aeruginosa, and C. albicans, which was found to be the best growth inhibition concentration. The DPPH scavenging activity shows a higher activity till the concentration of 64 µg/mL. The hemolytic activity of 25% is obtained at 128 µg/mL and below 64 µg/mL, there is no hemolytic activity. The gas chromatography-mass spectrometry (GC-MS) analysis of the secondary metabolites shows the presence of 17 compounds. Among them, there were four major compounds (i) cyclo(L-prolyl-L-valine) (probability of 95.63%), (ii) pyrrolo[1,2-a]pyrazine- 1,4-dione,hexahydro-3-(2-methylpropl) (probability of 94.45%), (iii) 2,5-piperazinedione,3,6-bis(2-methylpropyl) (probability of 71.94%) and (iv) pyrrolo[1,2-a]pyrazine-1,4-dione,hexahydro-3-(phenylmethyl) (probability of 88.01%). Conclusion In conclusion, the isolated bacterium is confirmed to be Halobacillus sp. and the secondary metabolites produced by this bacterium could be the potential source for the development of novel antimicrobial and antioxidant compounds that are highly biologically compatible. Further research may help to develop novel compounds in the pharmaceutical industry.