Extended-spectrum β-lactamase (ESBL)-producing bacteria pose a big challenge in clinical practices, warranting a new therapeutic strategy. In this study, methanol extract of the marine cyanobacterium Oscillatoria acuminata NTAPC05 was fractionated under bioassay guidance and the fractions were tested against three well-characterized ESBL-producing bacteria Escherichia coli U655, Stenotrophomonas maltophilia B929 and Enterobacter asburiae B938. Out of the four HPLC fractions, fraction 2 showed bactericidal activity against all the three ESBL producers much more efficiently (MIC 100 μg ml−1) than the fourth-generation cephalosporin (MIC >125 μg ml−1). The active fraction was subjected to time-kill test at concentrations of 1/2 × MIC, 1 × MIC and 2 × MIC, and the results substantiated the bactericidal property of the fraction against the ESBL producers. Spectral analysis revealed monogalactosyldiacylglycerol containing a palmitoyl (MGDG-palmitoyl), being reported for the first time, as the active fraction, and its bactericidal property against ESBL producers was determined. The active fraction appears to damage the bacterial membrane leading to lysis of the cell, as revealed in confocal laser scanning microscopy (CLSM) analysis, that was confirmed in scanning electron microscopic analysis. Cytotoxicity assay revealed the O. acuminata compound to be safe to a normal cell line HEK293 (human embryonic kidney cell). The in silico analysis of MGDG-palmitoyl revealed two successive H-bonding interactions with Leu198 of TEM1 β-lactamase. Taken together, the MGDG-palmitoyl from O. acuminata NTAPC05 offers potential to develop analogs as a therapeutic for bacteremia caused by ESBL producers.
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Bradford, P. A. Extended-spectrum β-lactamases in the 21st century: characterization, epidemiology, and detection of this important resistance threat. Clin. Microbiol. Rev. 14, 933–951 (2001).
Rishi, H. P. D. & John, C. ESBLs: a clear and present danger. Critical Care Res. Pract. 2012, 625170 (2012).
Sanders, C. C. Chromosomal cephalosporinases responsible for multiple resistances to newer beta-lactam antibiotics. Ann. Rev. Microbiol. 41, 573–593 (1987).
Paterson, D. L. & Bonomo, R. A. Extended-spectrum b-lactamases: a clinical update. Clin. Microbiol. Rev. 18, 657–686 (2005).
Bonnet, R. Growing group of extended-spectrum b-lactamases: the CTX-M enzymes. Antimicrob. Agents Chemother. 48, 1–14 (2004).
Albertini, M. T. et al. Surveillance of methicillin-resistant Staphylococcus aureus (MRSA) and Enterobacteriaceae producing extended-spectrum beta-lactamase (ESBLE) in Northern France: a five-year multicentre incidence study. J. Hosp. Infect. 52, 107–113 (2002).
Teng, C. P., Chen, H. H., Chan, J. & Lye, D. C. Ertapenem for the treatment of extended-spectrum beta-lactamase-producing gram-negative bacterial infections. Int. J. Antimicrob. Agents 30, 356–359 (2007).
Rasheed, M. U., Thajuddin, N., Ahamed, P., Teklemariam, Z. & Jamil, K. Antimicrobial drug resistance in strains of Escherichia coli isolated from food sources. Rev. Inst. Med. Trop. São Paulo 56, 341–346 (2014).
Tandeau-de-Marsac, H. J. Adaptation of cyanobacteria to environmental stimuli: new steps towards molecular mechanisms. FEMS Microbiol. Lett. 104, 119–190 (1993).
Mayer, A. M. S. & Hamann, M. T. Marine pharmacology in 2001–2002: marine compounds with anti helmintic, antibacterial, anticoagulant, anti diabetic, antifungal, anti-inflammatory, anti malarial, anti platelet, antiprotozoal, anti tuberculosis, and antiviral activities, affecting the cardiovascular, immune and nervous systems and other miscellaneous mechanisms of action. Comp. Biochem. Physiol. C 140, 265–286 (2005).
Moreau, J. D., Pasando, P., Bernand, P., Caram, B. & Pinnat, J. C. Seasonal variation in the production of antifungal substrates by dictyotales (brown algae) from the French Mediterranean coast. Hydobiologia 2, 1097–1132 (1988).
Renu, A. Antibacterial activities of freshwater algae Chlorella ellipsoidea. J. Basic Appl. Biol. 4, 22–26 (2010).
Abed, R. M. M. et al. Cyanobacterial diversity and bioactivity of inland hyper saline microbial mats from a desert stream in the Sultanate of Oman. Fottea 11, 215–224 (2011).
Rama murthy, V., Raveendran, S., Thirumeni, S. & Krishnaveni, S. Antimicrobial activity of heterocytic cyanobacteria. Int. J. Adv. Lif. Sci. 1, 32–39 (2012).
CLSI Performance Standards for Antimicrobial Susceptibility Testing: 22nd Informational Supplement M100-S22, (Clinical and Laboratory Standard Institute, Wayne, PA, (2012).
Desikachary, T. V. Cyanophyta, (ICAR Publication, New Delhi, (1959).
Smoker, J. A. & Barnum, S. R. Rapid small scale DNA isolation from filamentous cyanobacteria. FEMS Microbiol. Let. 56, 119–122 (2006).
Tamura, K. et al. MEGA5: molecular evolutionary genetics analysis using maximum likelihood evolutionary distance, and maximum parsimony methods. Mol. Biol. Evol. 28, 2731–2739 (2011).
Cheesbrough, M. inMedical Laboratory Manual for Tropical Countries Vol. 2, 2–392 (Tropical Health Technology Publications and Butterworth–Heinemann, Cambridge, UK, (2002).
Wikler, M. A. Performance Standards for Antimicrobial Susceptibility Testing; Eighteenth Informational Supplement; M100-S18. C.L.S.I. (Clinical and Laboratory Standard Institute), Pennsylvania, PA, USA 28, 46–52 (2008).
Weigand., I., Hilpert, R. & Hancock, R. E. W. Agar and broth dilution methods to determine the minimum inhibitory concentration (MIC) of antimicrobial substance. Nat. Protoc. 3, 163–175 (2008).
Eliopoulos, G. M. & Moellering, R. C. inAntibiotics in Laboratory Medicine 4thedn(ed.Lorain V) 330–396 The Williams & Wilkins, Baltimore, MD, USA, (1996).
Denizot, F. & Lang, R. Rapid colorimetric assay for cell growth and survival: modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J. Immunol. Methods 89, 271–277 (1986).
Ahmad, S. & Yadava, J. N. S. Rapid detection of b-lactam antibiotic resistance among clinical isolates of Escherichia coli. India Vet. Med. J. 3, 256–259 (1979).
Morris, G. M. et al. AutoDock4 and AutoDockTools4: automated docking with selective receptor flexibility. J. Comput. Chem. 30, 2785–2791 (2009).
Payne, D. J. Desperately seeking new antibiotics. Science 321, 1644–1645 (2008).
Lipsitch, M., Bergstrom, C. T. & Levin, B. R. The epidemiology resistance in hospital; Paradoxes and prescription. Proc. Natl Acad. Sci. USA 97, 1938–1943 (2000).
Pitout, J. D., Hanson, N. D. & Church, D. L. Population-based laboratory surveillance for Escherichia coli producing extended spectrum beta lactamases: Importance of community isolates with blaCTX-M genes. Clin. Infect. Dis. 38, 1736–1741 (2004).
Sanders, C. C. & Sanders, W. E. β-Lactamase resistance in Gram-negative bacteria: global trends and clinical impact. Clin. Infect. Dis. 15, 824–839 (1992).
Kerby, N. N. & Rowell, P. inPhotosynthetic Prokaryotes (eds Mann, N H & Car,N.G.) 233–254 (Plenum Press, New York, (1992).
Gupta, A. B. & Shrivastava, G. C. On antibiotic properties of some fresh water algae. Hydrobiologia 25, 285–288 (1965).
Schlegel, I., Doan, N. T., Chazal, N. & Smith, G. D. Antibiotic activity of new cyanobacterial isolates from Australia and Asia against green algae and cyanobacteria. J. Appl. Phycol. 10, 471–479 (1999).
Harriman, A. (Photo) isomerization dynamics of merocyanine dyes in solution. J. Photochem. Photobiol. A 65, 79–93 (1992).
Holzl, G. & Dormann, P. Structure and function of glycoglycerolipids in plants and bacteria. Prog. Lipid Res. 46, 225–243 (2007).
Heinz, E. & Roughan, P. G. Similarities and differences in lipid metabolism of chloroplasts isolated from 18:3 and 16:3 plants. Plant Physiol. 72, 273–279 (1983).
Dormann, P. & Benning, C. Galactolipids rule in seed plants. Trends Plant Sci. 7, 112–118 (2002).
Shirahashi, H. et al. Isolation and identification of anti-tumor-promoting principles from the fresh-water cyanobacterium Phormidium tenue. Chem. Pharm. Bull. 41, 1664–1666 (1993).
Morimoto, T. et al. Antitumor promoting glycerol-glycolipids from the green alga Chlorella vulgaris. Phytochemistry 40, 1433–1437 (1995).
Frentzen, M., Weier, D. & Feussner, I. Reports on symposia and congresses. Eur. J. Lipid Sci. Technol. 105, 784–792 (2003).
Vered, R. et al. New acylated sulfoglycolipids and digalactolipids and related known glycolipids from cyanobacteria with a potential to inhibit the reverse transcriptase of HIV-1. J. Nat. Prod. 60, 1251–1260 (1997).
Shai, Y. Mode of action of membrane active antimicrobial peptides. Biopolymers 66, 236–248 (2002).
Hameed, A. S. et al. In vitro antibacterial activity of ZnO and Nd doped ZnO nanoparticles against ESBL producing Escherichia coli and Klebsiella pneumoniae. Sci. Rep. 6, 24312 (2016).
Dhara, L., Tripathi, A. & Pal, A. Molecular characterization and in silico analysis of naturally occurring TEM β-lactamase variants among pathogenic Enterobacteriaceae infecting Indian patients. Biomed. Res. Int. 2013, 783540 (2013).
Bhagavat, R., Saqib, A. & Karigar, C. Molecular docking studies of novel palmitoyl- ligands for cyclooxygenase-2. Chem. Biol. Drug Des. 79, 1043–1048 (2012).
We are grateful to Dr Hemalatha Rao, Medwin Hospital (Hyderabad, India) for providing the clinical isolates. We thank the Deanship of Scientific Research at King Saud University for funding the work through the research group project (Ref No RGP-VPP-332). We thank DST- PURSE scheme (Project Ref No- SR/FT/LS-113/2009) of the Department of Science and Technology (DST), New Delhi, India, for the Confocal Laser Scanning Microscopy facility. This work was otherwise supported by the Project (Ref No BT/PR4815/AAQ/3/587/2012, BT/PR6619/PBD/26/310/2013, BT/IN/Indo-UK/SuBB/23/NT/2013 and BT/PR7005/PBD/ 26/357/2015) sanctioned by the Department of Biotechnology (DBT), New Delhi, India.
AP and NT performed most of the experiments and drafted the manuscript; MUR carried out the sample collection from humans and identified the bacterial strains; KPMN investigated the antibiotic resistance pattern and identified ESBL producers; NR contributed to the amplification of ESBL genes and interpretation of results; SS obtained the microphotograph and identification of cyanobacterium; MYMI contributed in 16S DNA amplification in ESBL producers; NSA, CA and SAA equally contributed in the analysis and interpretation of data; MAA carried out the cell line study, coordinated the manuscript drafting and language editing. All authors approved the final version of the manuscript.
The authors declare no conflict of interest.
Supplementary Information accompanies the paper on The Journal of Antibiotics website
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Parveez Ahamed, A., Rasheed, M., Peer Muhamed Noorani, K. et al. In vitro antibacterial activity of MGDG-palmitoyl from Oscillatoria acuminata NTAPC05 against extended-spectrum β-lactamase producers. J Antibiot 70, 754–762 (2017) doi:10.1038/ja.2017.40
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