Sclerotiorin: a Novel Azaphilone with Demonstrated Membrane Targeting and DNA Binding Activity against Methicillin-Resistant Staphylococcus aureus
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The emergence of multi-drug resistant, pathogenic methicillin-resistant Staphylococcus aureus (MRSA) is a threat to global health and has created a need for novel functional therapeutic agents. In this study, we evaluated the underlying mechanisms of the anti-MRSA effect of an azaphilone pigment, sclerotiorin (SCL) from Penicillium sclerotiorum. The antimicrobial activity of SCL was evaluated using agar disc diffusion, broth microdilution, time-kill assays and biophysical studies. SCL exhibits selective activity against Gram positive bacteria including MRSA (range, MIC = 128-1028 μg/ml) and exhibited rapid bactericidal action against MRSA with a > 4 log reduction in colony forming units within three hours of administration. Biophysical studies, using fluorescent probes and laser or electron microscopy, demonstrated a SCL dose-dependent alternation in membrane potential (62.6 ± 5.0.4% inhibition) and integrity (> 95 ± 2.3%), and the release of UV260 absorbing materials within 60 min (up to 3.2 fold increase, p < 0.01) of exposure. Further, SCL localized to the cytoplasm and hydrolyzed plasmid DNA. While in vitro checkerboard studies revealed that SCL potentiated the antimicrobial activity of topical antimicrobials such as polymixin, neomycin, and bacitracin (Fractional Inhibitory Concentration Index range, 0.26-0.37). Taken together these results suggest that SCL targets the membrane and DNA of MRSA to facilitate its anti- MRSA antimicrobial effect.
Abstract
The emergence of multi-drug resistant, pathogenic methicillin-resistant Staphylococcus aureus (MRSA) is a threat to global health and has created a need for novel functional therapeutic agents. In this study, we evaluated the underlying mechanisms of the anti-MRSA effect of an azaphilone pigment, sclerotiorin (SCL) from Penicillium sclerotiorum. The antimicrobial activity of SCL was evaluated using agar disc diffusion, broth microdilution, time-kill assays and biophysical studies. SCL exhibits selective activity against Gram positive bacteria including MRSA (range, MIC = 128-1028 μg/ml) and exhibited rapid bactericidal action against MRSA with a > 4 log reduction in colony forming units within three hours of administration. Biophysical studies, using fluorescent probes and laser or electron microscopy, demonstrated a SCL dose-dependent alternation in membrane potential (62.6 ± 5.0.4% inhibition) and integrity (> 95 ± 2.3%), and the release of UV260 absorbing materials within 60 min (up to 3.2 fold increase, p < 0.01) of exposure. Further, SCL localized to the cytoplasm and hydrolyzed plasmid DNA. While in vitro checkerboard studies revealed that SCL potentiated the antimicrobial activity of topical antimicrobials such as polymixin, neomycin, and bacitracin (Fractional Inhibitory Concentration Index range, 0.26-0.37). Taken together these results suggest that SCL targets the membrane and DNA of MRSA to facilitate its anti- MRSA antimicrobial effect.
참고문헌 (35)
- Stewart CM, "Staphylococcus aureus growth boundaries : moving mechanistic predictive mode is based on solute-specific effects", Appl. Environ. Microbiol, 68, pp.1864~1871 (2002)
- Tommasi R, "ESKAPEing the labyrinth of antibacterial discovery", Nat. Rev. Drug. Disc, 14, pp.529~542 (2015)
- Friedman M, "Antibiotic-resistant bacteria : prevalence in food and inactivation by food compatible compounds and plant extracts", J. Agric. Food. Chem, 63, pp.3805~3822 (2015)
- Osmanova N, "Azaphilones a class of fungal metabolites with diverse biological activities", Phytochem. Rev, 9, pp.315~342 (2010)
- Dong J, "New nematicidal azaphilones from the aquatic fungus Pseudohalonectria adversaria YMF1.01019", FEMS. Microbiol. Lett, 264, pp.65~69 (2006)
- Yoshida E, "Isolation of three new azaphilones, leteusins C, D, and E, from an ascomycete, Talaromyces luteus", Chem. Phar. Bull, 44, pp.284~287 (1996)
- Yang DJ, "New isochromophilones VII and VIII produced by Penicillium sp. FO4164", J. Antibiot, 49, pp.223~229 (1996)
- Yu BZ, "Phomoeuphorbins A-D, azaphilones from the fungus Phomopsis euphorbiae", Phytochemistry, 69, pp.2523~2526 (2008)
- Dos Santos P, "Antioxidant and antibacterial activities of the chlorine pigment sclerotiorin from Penicillium mallochii and its chemotaxonomic significance", Biochem. Syst. Ecol, 86, pp.103915~ (2019)
- Musso L, "Natural and semisynthetic azaphilones as a new scaffold for Hsp90 inhibitors", Bioorg. Med. Chem, 18, pp.6031~6043 (2010)
- Nam JY, "8-O-Methylsclerotiorinamine, antagonist of the Grb2-SH2 domain, isolated from Penicillium multicolor", J. Nat. Prod, 63, pp.1303~1305 (2000)
- Pairet L, "Azaphilones with endothelin receptor binding activity produced by Penicillium sclerotiorum : taxonomy, fermentation, isolation, structure elucidation and biological activity", J. Antibiot, 48, pp.913~923 (1995)
- Chidananda C, "Sclerotiorin, from Penicillium frequantans, a potent inhibitor of aldose reductase", Biotechnol. Lett, 28, pp.1633~1636 (2006)
- Tomoda H, "Structure-specific inhibition of cholesteryl ester transfer protein by azaphilones", J. Antibiot, 52, pp.160~170 (1999)
- Giridharan P, "Anticancer activity of sclerotiorin, isolated from endophytic fungus cephalotheca faveolata Yaguchi, Nishim, &Udagawa", Ind. J. Exp. Biol, 50, pp.464~468 (2012)
- Arunpanichlert J, "Azaphilone and isocoumarin derivatives from the endophytic fungus Penicillium Sclerotiorum PSU A-13", Chem. Pharm. Bull, 58, pp.1033~1036 (2010)
- Lin L, "Design, synthesis, and fungicidal activity of novel sclerotiorin derivatives", Chem. Biol. Drug. Des, 80, pp.682~692 (2012)
- Chen D, "Sclerotiorin inhibits protein Kinase G from Mycobacterium tuberculosis and impairs mycobacterial growth in macrophages", Tuberculosis, 103, pp.37~43 (2017)
- Gomes DC, "Sequential fungal fermentation biotransformation process to produce a red pigment from Sclerotiorin", Food Chem, 210, pp.355~361 (2016)
- Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; approved standards, "Clinical and Laboratory Standards Institute", Clinical and Laboratory and Standards Institute, ~, pp.CLSI document M45, 3rd Ed (2016)
- Pag U, "In vitro activity and mode of action of diasteriometic antimicrobial peptides against bacterial clinical isolates", J. Antimicrob. Chemother, 53, pp.230~239 (2004)
- Uppu DSSM, "Membrane active macromolecules resensitize NDM-1 gram negative clinical isolates to tetracycline antibiotics", PLoS One, 10, pp.e0119422~ (2015)
- Carson CF, "Mechanism of action of Melaleuca alternifolia(Tea tree oil)on Staphylococcus aureus determined by time kill assay, lysis, leakage, and salt tolerance assays, and electron microscopy", Antimicrob. Agents. Chemother, 46, pp.1914~1920 (2002)
- Tyagi P, "Bactericidal activity of curcumin I is associated with damaging of bacterial membrane", PLoS One, 10, pp.e0121313~ (2015)
- Nair DR, "Characterization of a novel small molecule that potentiates β-lactam activity against Gram-positive and Gramnegative pathogens", Antimicrob. Agents. Chemother, 59, pp.1876~1885 (2015)
- Song-lin Zhou, "Penicilazaphilone C, a new antineoplastic and antibacterial azaphilone from the Marine Fungus Penicillium sclerotiorum", Archives of Pharmacal Research, 39(12), pp.1621~1627 (2016), 대한약학회
- Lucas EMF, "Antimicrobial properties of Sclerotiorin, isochromophilone VI, and pencolide, metabolites, from a brazilian cerrado isolate of Penicillium sclerotiorum Van Beyma", Braz. J. Microbiol, 38, pp.785~789 (2007)
- Gao SS, "Comazaphilones A-F, azaphilone derivatives from the marine sediment-derived fungus Penicillium commune QSD 17", J. Nat. Prod, 74, pp.256~261 (2011)
- King JD, "Review : lipopolysaccharide biosynthesis in Pseudomonas aeruginosa", Innate Immun, 15, pp.261~312 (2009)
- Bell PJL, "Epicocconone, a novel fluorescent compound from the fungus Epicoccum nigrum", J. Am. Chem. Soc, 25, pp.9304~9305 (2003)
- Choi HY, "Epicocconone, a new cellpermeable long stoke’s shift fluorescent stain for live cell imaging and multiplexing", J. Fluoresc, 16, pp.475~482 (2006)
- Mitchell P, "Chemiosmotic coupling in oxidative and photosynthetic phosphorylation", Biol. Rev. Camb. Philos. Soc, 41, pp.445~502 (1966)
- Cheng MJ, "Secondary metabolites from the endophytic fungus Biscogniauxia formosana and their antimycobacterial activity", Phytochem. Lett, 5, pp.467~472 (2012)
- Farha MA, "Collapsing the proton motive force to identify synergistic combinations against Staphylococcus aureus", Cell. Chem. Biol, 20, pp.1168~1178 (2013)
- del Castillo FJ, "Construction and characterization of mutations at codon 751 of the Escherichia coli gyrB gene that confer resistance to the antimicrobial peptide microcin B17 and alter the activity of DNA gyrase", J. Bacteriol, 183, pp.2137~2140 (2001)