Identification of Multi-Potent Protein Subtilisin A from halophilic bacterium Bacillus firmus VE2.

Autor: P M; Department of Microbiology, Faculty of Science, Annamalai University, Chidambaram, 608002, Tamilnadu, India., Moopantakath J; Department of Genomic Science, Central University of Kerala, Tejaswini Hills, Periya, Kasaragod, 671320, Kerala, India., Imchen M; Department of Genomic Science, Central University of Kerala, Tejaswini Hills, Periya, Kasaragod, 671320, Kerala, India., Kumavath R; Department of Genomic Science, Central University of Kerala, Tejaswini Hills, Periya, Kasaragod, 671320, Kerala, India. Electronic address: RNKumavath@gmail.com., SenthilKumar PK; Department of Microbiology, Faculty of Science, Annamalai University, Chidambaram, 608002, Tamilnadu, India. Electronic address: drpks1980@gmail.com.
Jazyk: angličtina
Zdroj: Microbial pathogenesis [Microb Pathog] 2021 Aug; Vol. 157, pp. 105007. Date of Electronic Publication: 2021 May 25.
DOI: 10.1016/j.micpath.2021.105007
Abstrakt: Screening of halophiles with antimicrobial activity in saltpan soil samples from Nagapattinam district, Tamil Nadu, revealed isolate VE-2 as the most potent, identified as Bacillus firmus strain VE-2 through 16s rRNA gene sequencing. It had an optimum growth condition (OD 3.1) and antimicrobial protein (AMP) production (450 μg/mL) at 37 °C, pH 8, 25% NaCl, and 36 h incubation. SDS-PAGE analysis of the purified AMP showed the molecular weight of 36 kDa. HPLC analysis of the purified AMP showed different amino acids, such as asparagines, alanine, lysine, proline, threonine, glycine, cysteine, serine, aspartic acid leucine, and valine. Further characterization and identification using FT-IR, 2D-PAGE, MALDI-TOF, and in-silico analysis showed that the isolated AMP had the highest similarity to Subtilisin-A. It showed antibacterial activity against clinical bacterial pathogens like S. aureus, S. pyogenes, C. diphtheria, E. coli, and P. aeruginosa with the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration of 2.5 μg/mL and 20 μg/mL and also against various fungal pathogens such as A. niger, A. flavus, C. albicans, C. tropicalis and C. parapsilosis with the MIC and minimum fungicidal concentrations of 1.25-80 μg/mL. The purified AMP had excellent antioxidant potential, showed a scavenging effect against DPPH and Nitric oxide radicals, and displayed anticancer activity against HeLa cell lines with the IC 50 values 53 μg/mL. Hence, the purified bioactive antimicrobial peptides (AMP) could also be used in anticancer therapies.
(Copyright © 2021 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: