Purification and characterisation of a novel antistaphylococcal peptide (ASP-1) from Bacillus sp. URID 12.1
| Autor: | Utpal Roy, Sushma Nema, Ajay Ghosh Chalasani |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Microbiology (medical) Methicillin-Resistant Staphylococcus aureus Circular dichroism Spectrometry Mass Electrospray Ionization 030106 microbiology Peptide Bacillus subtilis Microbial Sensitivity Tests medicine.disease_cause 03 medical and health sciences Cell Line Tumor medicine Humans Pharmacology (medical) Peptide sequence chemistry.chemical_classification Chromatography Reverse-Phase Chromatography Molecular mass biology Chemistry General Medicine Hep G2 Cells biology.organism_classification 030104 developmental biology Infectious Diseases Biochemistry Staphylococcus aureus Acetylation Biofilms Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization MCF-7 Cells Peptides Lactone Antimicrobial Cationic Peptides |
| Zdroj: | International journal of antimicrobial agents. 51(1) |
| ISSN: | 1872-7913 |
| Popis: | A strong antistaphylococcal peptide (ASP-1) from Bacillus subtilis URID 12.1 strain that is active against cefoxitin- and methicillin-resistant Staphylococcus aureus clinical isolates was purified to homogeneity by solvent extraction, silica gel-based adsorption chromatography and reversed-phase high-performance liquid chromatography. The peptide sequence of ASP-1 as determined by MALDI-TOF/MS and ESI-FTICR-MS was acetylated Phe-Thr-Ala-Val-Dhb-Phe-Ile/Leu. The peptide was further analysed by alkaline hydrolysis, ESI-Q-TOF-MS and an ion mobility assay, which detected the presence of a lactone ring in the intact peptide and a cyclic nature, subsequently revealing the linearised peptide sequence as acPhe-Leu-Phe-Thr-Val-Ala-Dhb. Based on the molecular mass (804.5 Da), peptide sequence and amino acid composition, ASP-1 was identified as a lactone ring-containing peptide similar to TL-119, a poorly studied cyclic depsipeptide. Circular dichroism spectroscopy revealed its predominantly random structure in aqueous solution and its β-sheet conformation in methanol. Minimum inhibitory concentrations (MICs) of the purified peptide against S. aureus and methicillin-resistant S. aureus (MRSA) ranged from 2 µg/mL to 64 µg/mL. At sub-MICs and 1× MIC, ASP-1 showed a strong antibiofilm characteristic. ASP-1 at a concentration of 128 µg/mL did not show haemolytic activity, and no cytotoxicity was observed against hepatic carcinoma and breast carcinoma cell lines at the same concentration. Peptide ASP-1 with anti-MRSA and antibiofilm abilities and non-haemolytic and non-cytotoxic properties has not been reported previously. These findings suggest that it may serve as a lead molecule for developing alternative topical antibacterial agents. |
| Databáze: | OpenAIRE |
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