PRODUCTION, PURIFICATION, AND CHARACTERIZATION OF BIOACTIVE METABOLITES PRODUCED FROM RARE ACTINOBACTERIA PSEUDONOCARDIA ALNI

Rabab Omran; Mohammed Fadhil Kadhem

doi:10.22159/ajpcr.2016.v9s3.14961

Authors

Rabab Omran babylon university http://orcid.org/0000-0001-9449-3421
Mohammed Fadhil Kadhem

DOI:

https://doi.org/10.22159/ajpcr.2016.v9s3.14961

Abstract

ABSTRACT
Objectives: Pseudonocardia alni exhibits antimicrobial activity against tested pathogenic Staphylococcus aureus, Microsporum canis, and Trichophyton
mentagrophyte. The present paper aimed to optimize various cultural conditions for antimicrobial metabolite production, purification, and
characterization of the active substance.
Methods: The effects of various parameters such as culture media, carbon and nitrogen sources, phosphate concentration, pH, temperature, incubation
period, and agitation rate on bioactive metabolite production were studied using a flask scale with varying single parameter. The active substances
were purified by adsorption chromatography using Silica gel column and Sephadex LH 20 column, and the physical, chemical, and biological properties
were characterized.
Results: The metabolite production by P. alni was greatly influenced by various cultural conditions. It produced high levels of the antimicrobial
substance in International Streptomyces project-2 broth compared with that in potato dextrose broth. The optimum parameters for antimicrobial
production from the actinobacterium occurred in the production medium consisting of glucose (1%) and tryptone (1%), 0.001 M of K2HPO4 and
0.05M glycine at initial pH 8.5 and incubated at 30Â°C for 4 d in stand incubator. The higher concentration of phosphate buffer salts (Ëƒ0.01M) repressed
the bioactive production. The purified active substance had relative factor Rf=0.53 in the mobile phase of a thin layer chromatography system and
the maximum absorbance (Î» max) at 216 nm. The results of infrared spectra Fourier transform-infrared spectroscopy analysis indicate that it may be
regarded to glycopeptide antibiotic. The purified substance had antibacterial and antifungal activities as well as cytotoxic activity in breast cancer cell
line Michigan Cancer Foundation-7 and normal hepatic cell line (WRL-68) at a percentage up to 23.7% and 7.64%, respectively.
Conclusions: The actinobacterium P. alni was a novel strain having the ability to produce antimicrobial and anticancer substances.
Keywords: Pseudonocardia alni, Antifungalâ€“antibacterial, Anticancer optimization, Production, Purification, Characterization.

Downloads

Download data is not yet available.

Author Biography

Rabab Omran, babylon university

biology deparament , biotechnology-genetic engineering

References

Newman DJ, Cragg GM. Natural products as sources of new drugs over the last 25 years. J Nat Prod 2007;70(3):461-77.

Gehlot V, Mahant S, Das K, Das R. Most of the Helicobacter pylori isolates are resistant to levofloxacin in north India. Int J Pharm Pharm Sci 2016;8(1):454-6.

Gehlot V, Mahant S, Vijayraghwan P, Das K, Hoda S, Das R. Therapeutic potential of lichen Parmeliaperlata against dual drag-resistant Helicobacter pylori isolates. Int J Pharm Pharm Sci 2016;8(1):205-8.

BÃ©rdy J. Bioactive microbial metabolites. J Antibiot (Tokyo) 2005;58(1):1-26.

da Silva IR, Martins MK, Carvalho CM, de Azevedo JL, de Lima ProcÃ³pio RE. The effect of varying culture conditions on the production of antibiotics by Streptomyces spp., isolated from the Amazonian soil. Ferment Technol 2012;1:105.

Usha Kiranmayi M, Sudhakar P, Sreenivasulu K, Vijayalakshmi M. Optimization of culturing conditions for improved production of bioactive metabolites by Pseudonocardia sp. VUK-10. Mycobiology 2011;39(3):174-81.

Omran R, Kadhem MF. Isolation and identification of antibiotic-producing actinomycetes. J Babylon Univ 2012;22(1):285-92.

Shirling EB, Gottlieb D. Methods for characterization of Streptomyces species. Int J Sys Bacteriol 1966;16(3):313-40.

Perez C, Pauli M, Bazevque P. An antibiotic assay by the agar-well diffusion method. Acta Biol Med Exp 1990;15:113-5.

Pavia DL, Lampman GM, Kriz GS, Vyvyan JR. Microscale andMacroscale Techniques in the Organic Laboratory. USA: Brooks/Cole; 2002.

Mohanty S, Basu S. Fundamentals of Practical Clinical Biochemistry. New Delhi. India: BI Publications Pvt.; 2006. p. 282.

Augustine SK, Bhavsar SP, Baserisalehi M, Kapadnis BP. A non-polyene antifungal antibiotic from Streptomyces albidoflavus PU 23. Indian J Exp Biol 2006;42:928-32.

Freshney RI. The Culture of an Animal Cell. 6th ed. New York: Wily-Liss; 2010.

Chih PL, Wei JT, Yuang LL, Yuh CK. The extracts from Nelumbo nucifera suppress cell cycle progression, cytokine genes expression, and cell proliferation in human peripheral blood mononuclear cells. Life Sci 2004;75:699-16.

Omran R. Corrosive lesions at concrete infrastructures as promising source for isolating bioactive actinobacteria. Am J Life Sci 2015;3(4):247-56.

Ripa FA, Nikkon F, Zaman S, Khondkar P. Optimal conditions for antimicrobial metabolites production from a new Streptomyces sp. Rupa-08pr isolated from Bangladeshi soil. Mycobiology 2009;37(3):211-4.

Raytapadar S, Paul AK. Production of an antifungal antibiotic by Streptomyces aburaviensis IDA-28. Microbiol Res 2001;155(4):315-23.

Aharonowitz Y, Demain AL. Influence of inorganic phosphate and organic buffers on cephalosporin production by Streptomyces clavuligerus. Arch Microbiol 1977;115(2):169-73.

Alduina R, Lo Piccolo L, Dâ€™Alia D, Ferraro C, Gunnarsson N, Donadio S, et al. Phosphate-controlled regulator for the biosynthesis of the dalbavancin precursor A40926. J Bacteriol 2007;189(22):8120-29.

Demain AL, Matteo CC. Phenylalanine stimulation of gramicidin S formation. Antimicrob Agents Chemother 1976;9(6):1000-03.

Thumar JT, Dhulia K, Singh SP. Isolation and partial purification of an antimicrobial agent from halotolerant alkaliphilic Streptomyces aburaviensis strain Kut-8. World J Microbiol Biotechnol 2010;26:2081-7.

Fiedler H, Dieter A, Hamm A, Goodfellow M, Muller WE, Brun R, et al. Pyrocoll an antibiotic, antiparasitic and antitumor compound produced by a novel alkaliphilic Streptomyces strain. J Antibiot 2003;56(7):639-46.

El-Naggar MY, Hassan MA, Said WY, El-Aassar SA. Effect of support materials on antibiotic MSW2000 production by immobilized Streptomyces violatus. J Gen Appl Microbiol 2003;49(4):235-43.

Miyoshi T, Iseki M, Konomi T, Imanaka H. Biosynthesis of bicyclomycin. I. Appearance of aerial mycelia negative strains (am-). J Antibiot (Tokyo) 1980;33(5):480-7.

Traub WH, Leonhard B. Heat stability of the antimicrobial activity of sixty-two antibacterial agents. J Antimicrob Chemother 1995;35(1):149-54.

Oskay M. Isolation and purification of two metabolites (KGG32-A & KGG32-B) from a soil bacterium, Streptomyces sp., KGG32. Int J Agric Biol 2011;13(3):369-74.

Lorian V. Antibiotics in Laboratory Medicine. USA: Lippincott Williams and Wilkins; 2005. p. 654.

Zhang C, Li S, Tian X, Niu S, Zhang W, Chen Y, et al. Pseudonocardians AC, new diaza anthraquinone derivatives from a deep-sea actinomycete Pseudonocardia sp. SCSI 01299. Mar Drugs 2011;9(8):1428-39.

Pavia DL, Lampman GM, Kriz GS, Vyvyan JR. Introduction to Spectroscopy. USA: Brooks/Cole; 2009.

Takeuchi M, Enokita R, Okazaki T, Kagasaki T, Inukai M. Helvecardins A and B, novel glycopeptide antibiotics I. Taxonomy, fermentation, isolation and physico-chemical properties. J Antibiot (Tokyo) 1991;4(3):263-70.

Jeena K, Liju VB, Kuttan R. Antitumor and cytotoxic activity of ginger essential oil (Zingiber officinale roscoe). Int J Pharm Pharm Sci 2015;7(8):341-44.

Thanekar DR, Tupe P, Dhodi J, Juvekar AR. Evaluation of in vitro cytotoxic activity of petroleum ether and methanol extract of Mentha arvensis (whole plant) on human cancer cell lines. Int J Pharm Pharm Sci 2014;6(8):169-72.