Production Purification And Biochemical Characterization Of Extracellular Pigment S From Actinomycetes Isolated From Rhizosphere Soil Of MP Regions India

Abstract

newlineActinomycetes, a large group of filamentous bacteria account for 70-80% of secondary metabolites available commercially. The present investigation was undertaken with an aim to identify and characterize pigment from actinomycetes. Actinomycetes were isolated from rhizosphere soil samples collected from different regions of Madhya Pradesh state. Out of 85 actinomycetes, only seven actinomycetes showed pigment production and based on diffusible pigment production ability one actinomycete ARITM02 was selected. The selected actinomycetes ARITM02 proceed for antimicrobial activity. Primary screening was done by agar overlay method and secondary screening was done by well agar diffusion method. The selected isolate showed antimicrobial activity against bacteria, fungi and molds but no activity against dermatophytes. newline The selected isolate was characterized by morphological, biochemical, physiological and molecular characterization and based on 16s rRNA sequencing the isolate confirmed as Strptomyces flaviofuscus ARITM02. The optimization of cultural parameters was done by various parameters like incubation time, temperature, pH, carbon sources, and nitrogen sources. The result of study showed 96 hours as best incubation time, 35 ºC best temperature, 7.5 pH was resulted best pH for growth of isolate, starch best carbon source and peptone best nitrogen source. newlineThe extraction of pigment was done by solvent extraction method using methanol. Thin layer chromatography was used for fractionation of culture extracts. The extracted crude compound further proceeds for thin layer chromatography and Rf value found 0.76. The pigment now purified by column chromatography and stored in ampoule. newline The pigment was characterized by UV-Vis spectroscopy which showed the lamda maximum of 433.23 FTIR spectroscopy suggested various functional groups like amino group, amide group, hydroxide, benzene and lactone group. The Mass spectroscopy and NMR spectroscopy showed that the molecular mass of pigment is 621.7 and molecular formula is C34H43