Anti-MRSA activity of a bioactive compound produced by a marine Streptomyces and its optimization using statistical experimental design

Document Type : Original Article

Authors

1 Department of Biology, University of Isfahan, Isfahan, Iran

2 Biotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran

Abstract

Objective(s): To address the alarming problem of methicillin-resistant Staphylococcus aureus (MRSA), herein, a marine Streptomyces capable of producing an anti-MRSA compound has been studied.
Materials and Methods: Strain MN41 was morphologically and physiologically characterized and then, molecularly identified using 16SrRNA analysis. To produce the bioactive compound in large scale, a kind of submerged liquid fermentation was adopted. The antibacterial agent was purified using a silica gel column followed by a semi-preparative HPLC and the isolated metabolite was identified using mass spectrometry, Nuclear magnetic resonance (NMR) and Fourier-transform infrared (FTIR).  Finally, the production process was subjected to a two steps optimization using Plackett-Burman design (PBD) and Response Surface Method (RSM), respectively. In addition, the antitumor activity of the active agent was studied.
Results: The purified compound with a molecular weight of 421.2 was identified as a natural pyrrole-derivative. The optimization revealed a significant effect for starch, pH, calcium carbonate and peptone on the production of this anti-MRSA compound and resulted in a 218% increase in the production yield.
Conclusion: The isolated pyrrole-derivative showed a remarkable activity against MRSA and also showed some promising anti-tumor activity.

Keywords

Main Subjects


1.  Maragakis LL, Perencevich EN, Cosgrove S. Clinical and economic burden of antimicrobial resistance. Expert Rev Anti Infect Ther 2008; 6:751-763.
2. Loomba PS, Taneja J, Mishra B. Methicillin and Vancomycin resistant S. aureus in hospitalized patients. J Glob Infect Dis 2010; 2:275-283.
3. Enright MC. The evolution of a resistant pathogen-the case of MRSA. Curr Opin Pharmacol 2003; 3:474-479.
4. Subramani R, Aalbersber W. Marine actinomycetes: an ongoing source of novel bioactive metabolites. Microbiol Res 2012; 167:571-580.
5. Dharmaraj S. Marine Streptomyces as a novel source of bioactive substances. World J Microbiol Biotechnol 2010; 26:2123-2139.
6. Arasu MV, Duraipandiyan V, Ignacimuthu S. Antibacterial and antifungal activities of polyketide metabolite from marine Streptomyces sp. AP-123 and its cytotoxic effect. Chemosphere 2013; 90:479-487.
7. Jang KH, Nam SJ, Locke JB, Kauffman CA, Beatty DS, Paul LA, et al. Anthracimycin, a potent anthrax antibiotic from a marine-derived actinomycete. Angew Chem In Ed Engl 2013; 52:7822-7824.
8. Jiao W, Zhang F, Zhao X, Hu J, Suh JW. A novel alkaloid from marine-derived Streptomyces xinghaiensis with broad-spectrum antibacterial and cytotoxic activities. PloS one 2013; 8:e75994.
9. De Carvalho CCCR, Fernandes P. Production of metabolites as bacterial responses to the marine environment. Mar Drugs 2010; 8:705-727.
10. Lam KS. Discovery of novel metabolites from marine actinomycetes. Curr Opin Microbiol 2006; 9:245-251.
11. Mohseni M, Norouzi H, Hamedi J, Roohi A. Screening of antibacterial producing actinomycetes from sediments of the Caspian Sea. Int J Mol Cell Med 2013; 2:64-71.
12. Rajeswari P, Jose PA, Amiya R, Jebakumar SRD. Characterization of saltern based Streptomyces sp. and statistical media optimization for its improved antibacterial activity. Front Microbiol 2015; 5:753.
13. Arul Jose P, Satheeja Santhi V, Jebakumar SR. Phylogenetic-affiliation, antimicrobial potential and PKS gene sequence analysis of moderately halophilic Streptomyces sp. inhabiting an Indian saltpan. J Basic Microbiol 2011; 51:348-356.
14. Greasham RL. Media for microbial fermentations. 2 st ed. Biotechnology Set; 1983.p. 127-139.
15. Wang Y, Fang X, An F, Wang G, Zhang X. Improvement of antibiotic activity of Xenorhabdus bovienii by medium optimization using response surface methodology. Microb Cell Fact 2011; 10:98.
16. Kanmani P, Karthik S, Aravind J, Kumaresan K. The use of response surface methodology as a statistical tool for media optimization in lipase production from the dairy effluent isolate Fusarium solani. ISRN Biotechnol 2012; 8:201.
17. Jose PA, Jebakumar SRD. Diverse actinomycetes from Indian coastal solar salterns-a resource for antimicrobial screening. J Pure Appl Microbiol 2013; 7:2569-2575.
18. Shirling EB, Gottlieb D. Methods for characterization of Streptomyces species. Int J Syst Evol Microbiol 1966; 16:313-340.
19. Stephen JR, McCaig AE, Smith Z, Prosser J, Embley TM. Molecular diversity of soil and marine 16S rRNA gene sequences related to beta-subgroup ammonia-oxidizing bacteria. Appl Environ Microbiol 1996; 62:4147-4154.
20. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kuma S. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 2011; 28:2731-2739.
21. Wu Z, Li S, Li J, Chen Y, Saurav K, Zhang Q, et al. Antibacterial and cytotoxic new napyradiomycins from the marine-derived Streptomyces sp. SCSIO 10428. Mar Drugs 2013; 11:2113-2125.
22. Saraswathi K, Sabitha Rani AM, Sindhu S, Arumugam P. Isolation, characterization of bioinspired secondary metabolites producing actinomycetes from marine soil samples. Int J Curr Microbiol Appl Sci 2015; 4:107-119.
23. Ebaba SS, Edrada RA, Lin W, Prokesh P. Methods for isolation, purification and structural elucidation of bioactive secondary metabolites from marine invertebrates. Nat Protoc 2008; 3:1820-1831.
24. Oniszczuk A, Wójtowicz A, Oniszczuk T, Olech M, Nowak R, Wojtunik K, et al. Extruded corn gruels containing linden flowers: quantitation of phenolic compounds and selected quality characteristics. Open Chem 2015;13:1209-1217.
25. Soheili V, Khedmatgozar Oghaz N, Sabeti Z, Fazly Bazzaz BS. The novel effect of cis-2-decenoic acid on biofilm producing Psudomonas aeruginosa. Microbiol Res (Pavia) 2016; 6:6158.
26. Ravikumar S, Fredimoses M, Gnanadesigan M. Anticancer property of sediment actinomycetes against MCF-7 and MDA-MB-231 cell lines. Asian Pac J Trop Biomed 2012; 2:92-96.
27. Zheng Z, Zeng W, Huang Y, Yang Z, Li J, Cai H, Su W. Detection of antitumor and antimicrobial activities in marine organism associated actinomycetes isolated from the Taiwan Strait. FEMS Microbiol lett 2000; 188:87-91.
28. Norouzi H, Danesh A, Mohseni M, Rabbani Khorasgani M. Marine Actinomycetes with probiotic potential and bioactivity against Multidrug-resistant bacteria. Int J Mol Cell Med 2018; 7:44-52.
29. Boyle VJ, Fancher ME, Ross RW. Rapid modified Kirby-Bauer susceptibility test with single, high-concentration antimicrobial disks. Antimicrob Agents Chemother 1973; 3:418-424.
30. Marzban A, Ebrahimipour G, Danesh A. Bioactivity of a novel glycolipid produced by a halophilic Buttiauxella sp. and improving submerged fermentation using a response surface method. Molecules 2016; 21:1256.
31. Jose PA, Sivakala KK, and Jebakumar SRD. Formulation and statistical optimization of culture medium for improved production of antimicrobial compound by Streptomyces sp. JAJ06. Int J Microbiol 2013; 2013:526260.
32. Kannan RR, Iniyan AM, Prakash VS. Isolation of a small molecule with anti-MRSA activity from a mangrove symbiont Streptomyces sp. PVRK-1 and its biomedical studies in Zebrafish embryos. Asian Pac J Trop Biomed 2011; 1:341-347.
33. Chaston JM, Suen G, Tucker SL, Andersen AW, Bhasin A, Bode E, et al. The entomopathogenic bacterial endosymbionts Xenorhabdus and Photorhabdus: convergent lifestyles from divergent genomes. PloS one 2011; 6:e27909.
34. Williams STM, Goodfellow M, Alderson G, Wellington EMP, Sneath PHA, Sackin MJ. Numerical classification of Streptomyces and related genera. J Gen Microbial 1983; 129:1743-1813.
35. Mousumi D, Dayanand A. Production and antioxidant attribute of L-glutaminase from Streptomyces enissocaesilis DMQ-2. Int J Latest Res Sci Technol 2013; 2:1-9.
36. Kemung HM, Tan LT, Khan TM, Chan KG, Pusparajah P, Goh BH, et al . Streptomyces as a prominent resource of future Anti-MRSA drugs. Front Microbiol 2018; 9:2221-2246.
37. Haste NM, Thienphrapa W, Tran DN, Loesgen S, Sun P, Nam SJ, et al. Activity of the thiopeptide antibiotic nosiheptide against contemporary strains of methicillin-resistant Staphylococcus aureus. J Antibiot 2012; 65:593-598.
38. Park HB, Le JK, Lee KR, Kwon HC. Angumycinones, A and B, two new angucyclic quinones from Streptomyces sp. KMC004 isolated from acidic mine drainage. Tetrahedron Lett 2014; 55:63-66.
39. CLSI. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically,; Tenth Edition: Approved Standard; 2015.M07-A10.Wayne, PA:CLSI.Available online at: http://www.facm.ucl.ac.be/intranet/CLSI/CLSI-2015-M07-A10-original.pdf
40. Li XB, Qiao B, Yuan YJ. Differential analysis of secondary metabolites by LC-MS following strain improvement of Streptomyces lydicus AS 4.2501. Biotechnol Appl Biochem 2006; 45:107-118.
41. Wilkerson WW, Copeland RA, Covington M, Trzaskos JM. Anti-inflammatory 4,5-diarylpyrroles activity as a function of cyclooxygenase inhibition. J Med Chem 1995; 38:3895-3901.
42. Lee H, Lee J, Lee S, Shin Y, Jung W, Kim JH, et al. A novel class of highly potent, selective, and non-peptidic inhibitor of ras farnesyl transferase (FTase). Bioorg Med Chem Lett 2001; 11:3069-3072.
43. Hughes CC, Kauffman CA, Jensen PR, Fenical W. Structures, reactivities, and antibiotic properties of the marinopyrroles A-F. J Org Chem 2010; 75:3240-3250.
44. Suffness M, Pezzuto JM. Assay related to cancer drug discovery. In: Hostettman K. editors. Methods in plant biochemistry: assay for bioactivity. London: Academic Press ;1990. p.71-133.
45. Praveen V, Ripathi D, Tripathi CKM, Bihari V. Nutritional regulation of actinomycin-D production by a new isolate of Streptomyces sindenensis using statistical methods. Indian J Exp Biol 2008; 46:138-144.
46. Marques DAV, Cunha MNC, Araújo JM, Lima-Filho JL, Converti A, Pessoa-Jr A, et al. Optimization of clavulanic acid production by Streptomyces daufpe 3060 by response surface methodology. Braz J Microbiol 2011; 42:658-667.
47. Guo Z, Shen L , Ji Z, Wu W. Enhanced production of a novel cyclic hexapeptide antibiotic (NW-G01) by Streptomyces alboflavus 313 using response surface methodology. Int J Mol Sci 2012; 13:5230-5241.
48. Raytapadar S, Paul AK. Production of an antifungal antibiotic by Streptomyces aburaviensis IDA-28. Microbiol. Res 2001; 155:315-323.