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Isolation and purification of antibacterial compound from Streptomyces levis collected from soil sample of north India.

ABSTRACT: During the screening programme for microbial cultures producing antimicrobial agents, an active microbial strain of Streptomyces was isolated from the agricultural soil of Narnaul, Haryana India. Physiological, biochemical characteristics and 16S ribosomal RNA sequence homology studies revealed that it was similar to Streptomyces levis (sequence similarity 100%). The microbial strain was submitted to Genomebio Technologies Pvt. Ltd., Pune, Maharashtra, India under Accession No. EU124569. The isolated strain was found to produce extracellular active compound showing strong antimicrobial activity against Klebsiella pneumoniae MTCC 109, Pseudomonas aeruginosa MTCC 741 and Staphylococcus aureus MTCC 96. The antibacterial compound was successfully isolated and purified. Structure elucidation of antibacterial metabolite with EI-MS/ HRMS showed molecular ion peak at m/z 686 [M+H]+. Whereas, elemental analysis of the said compound showed C = 61.31, H = 8.61, N = 2.04 and O = 28.02, and indicated a molecular formula of C35H59NO12. The presence of 'chromone' nucleus in the compound's chemical structure was confirmed by using 1HNMR studies. The present study reports the purification of potential antibacterial compound from Streptomyces levis isolated from the unexplored soil of north India and warrants for further characterization of this potential compound for optimum utilization for antimicrobial purposes.

SUBMITTER: Singh V

PROVIDER: S-EPMC6039041 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Isolation and purification of antibacterial compound from Streptomyces levis collected from soil sample of north India.

Singh Vineeta V Haque Shafiul S Khare Shruti S Tiwari Anil Kumar AK Katiyar Diksha D Banerjee Bikram B Kumari Krishna K Tripathi C K M CKM

PloS one 20180710 7

During the screening programme for microbial cultures producing antimicrobial agents, an active microbial strain of Streptomyces was isolated from the agricultural soil of Narnaul, Haryana India. Physiological, biochemical characteristics and 16S ribosomal RNA sequence homology studies revealed that it was similar to Streptomyces levis (sequence similarity 100%). The microbial strain was submitted to Genomebio Technologies Pvt. Ltd., Pune, Maharashtra, India under Accession No. EU124569. The iso ...[more]

PMID: 29990382

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Project description:BackgroundThe search for novel antibiotics continues to be of immense importance in research programs around the world for pharmaceutical, industrial and agricultural applications. Filamentous soil bacteria, belonging to the Streptomyces genus, are widely used as an important biological tool for their ability to produce a wide range of novel secondary metabolites, such as "antibiotics".ObjectivesThe aim of the present study was to isolate and identify a strain of Streptomyces with high antibiotic production capability.Materials and methodsThe soil sample was collected randomly from the agricultural land of Kushalnagar Taluk of Kodagu district, Karnataka, India. The ACTK2 strain was isolated by serial dilution method and identified based on cultural, morphological, microscopic, biochemical and sequence analysis of 16S rRNA gene parameters. The isolated ACTK2 was analyzed for antimicrobial activities by perpendicular streak and disc diffusion methods, against the Gram-positive bacteria Staphylococcus aureus (MTCC 96), Bacillus subtilis (MTCC 121), Gram-negative Escherichia. coli (MTCC 729), Enterococcus aerogenes (MTCC 2829) and filamentous fungi (Trichoderma harizianum (MTCC6046), Fusarium proliferatum (MTCC 9375). Further, an antimicrobial metabolite from the ACTK2 strain was extracted by solvent extraction method, using n-butanol. The production of the antimicrobial compound by the ACTK2 strain was optimized by using different nutritional media and cultural conditions.ResultsThe strain Streptomyces flavogriseus designated as ACTK2 (Accession number KC990785) isolated from the soil sample of Kushalnagar Taluk, Kodagu, Karnataka, India, exhibited a broad spectrum of antimicrobial activity against test microorganisms. The optimum growth and antimicrobial compound production by strain ACTK2 was found to be a maximal pH 8, in the shaker incubator at 28ºC, for a period of 10 days.ConclusionsThe crude n-butanol extract of the ACTK2 strain of S. flavogriseus showed a broad spectrum of antimicrobial activities against the test organisms and this opened further research investigations on purification and structural characterization of the active compounds from the crude extract.

Project description:The quest for novel broad spectrum bioactive compounds is needed continuously because of the rapid advent of pathogenic multi drug resistant organisms. Actinomycetes, isolated from unexplored habitats can be a solution of this problem. The motive of this research work was isolation of actinomycetes having potential antimicrobial activities from unexplored regions of Devbag and Tilmati beach. The isolated actinomycetes were screened against pathogenic microbes for antimicrobial activities through cross streak method. Enzyme production activity was checked for these actinomycetes for amylase, protease, cellulase and lipase enzymes. Further antimicrobial activity of ethyl acetate extract of the potent strain KS46 was performed. The strain KS46 was identified with 16S rRNA gene sequencing and secondary structure was analysed. Gas chromatography-Mass spectrometry (GC-MS) profiling was conducted to ascertain the presence of bioactive metabolites in the ethyl acetate extract. The collected samples were pre-treated and 70 actinomycetes were isolated. The Streptomyces sp. strain KS46 showed the best antimicrobial activity in primary screening. Ethyl acetate extract of the strain KS46 revealed antimicrobial activity against S. aureus, B. subtilis, B. cereus, E. faecalis, K. pneumoniae, E. coli, S. flexneri, C. albicans and C. glabrata. The 16S rRNA gene sequencing identified the strain KS46 as Streptomyces levis strain KS46. The GC-MS metabolite profiling of the ethyl acetate extract revealed the availability of 42 compounds including fatty acid esters, fatty acid anhydrides, alkanes, steroids, esters, alcohols, carboxylic ester, etc. having antibacterial, antifungal, antiproliferative, antioxidant activities. This study indicated that Devbag and Tilmati beaches being untapped habitats have enormous diversity of promising antimicrobial metabolite producing actinomycetes. Therefore, further exploration should be carried out to characterize the potential actinomycetes, which can be optimistic candidates for generation of novel antimicrobial drugs.

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Isolation and purification of antibacterial compound from Streptomyces levis collected from soil sample of north India.

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Isolation and purification of antibacterial compound from Streptomyces levis collected from soil sample of north India.

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