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Studies on the production and purification of an antimicrobial compound and taxonomy of the producer isolated from the marine environment of the Sundarbans

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Abstract

A microorganism isolated from the Sundarbans region of the Bay of Bengal, India, showed potent antimicrobial activity against gram-positive and gram-negative bacteria, molds, yeast and several multiple-drug-resistant (MDR) bacteria, including methicillin-resistant Staphylococcus aureus (MRSA). The isolate grew in the presence of 20% (w/v) NaCl, antibiotic production being maximum with 5% (w/v) NaCl in the production medium. Natural seawater stimulated antibiotic biosynthesis. The absence of catabolite repression during the synthesis of the antimicrobial substance was demonstrated by the utilization of glucose by this isolate. The 16S rRNA gene of this aerobic, gram-positive, mycelium- and spore-forming microorganism was amplified, and molecular phylogenetic analysis of the DNA sequence showed less than 93% similarity with its closest relative, indicating differentiation at the genus level. The highly stable, active principle was purified by butyl acetate extraction and silica-gel chromatography and a single compound was found to posses the broad-spectrum activity. Molecular characterization showed that the active compound is a lipid. Bioreactor studies demonstrated that antibiotic production is strongly dependent on the scale of operation and there is a definite relation between the dissolved oxygen concentration, medium pH, glucose utilization, cell differentiation and antibiotic production. Maximum production in 30 h could be obtained by regulation of the medium pH in the alkaline range by a combination of controlled addition of NaOH, regulation of the air supply and changes in the reactor configuration. Considering all of the above evidences and based on comparison with the current literature, a novel antimicrobial appears to have been isolated.

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Acknowledgments

Financial support from the All India Council for Technical Education (Grant no. 8019/RDII/R&D/BIO(135)200-01), Department of Science and Technology, New Delhi (Grant no. SR/FTP/LS-A-016/2001) to J. Mukherjee and Indian Council of Medical Research fellowship (No. 45/26/2001/Pharma/BMS) to M. Saha is thankfully acknowledged. Special thanks to Dr T. Sen for help during the toxicity tests.

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Correspondence to J. Mukherjee.

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Saha, M., Ghosh, D., Ghosh, D. et al. Studies on the production and purification of an antimicrobial compound and taxonomy of the producer isolated from the marine environment of the Sundarbans. Appl Microbiol Biotechnol 66, 497–505 (2005). https://doi.org/10.1007/s00253-004-1706-3

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  • DOI: https://doi.org/10.1007/s00253-004-1706-3

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