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KMID : 0545120100200111513
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Journal of Microbiology and Biotechnology
2010 Volume.20 No. 11 p.1513 ~ p.1520
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Purification and physiochemical characterization of melanin pigment from Klebsiella sp. GSK
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Shrishailnath S Sajjan
Guruprasad Kulkarni
Veeranagouda Yaligara
Lee Kyoung
T. B. Karegoudar
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Abstract
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The bacterium capable of producing melanin pigment in the presence of L-tyrosine was isolated from crop field soil sample and identified as Klebsiella sp. GSK based on morphological, biochemical and 16S rDNA sequencing. The polymerization of this pigment occurs outside the cell wall, which has granular structure as melanin ghosts. The chemical characterization of pigment particles showed acid resistant, alkali soluble, insoluble in most of the organic solvents and water. The pigment gets bleached when subjected to the action of oxidants as well as reductants. This pigment was precipitated with FeCl3, ammoniacal silver nitrate and potassium ferricynide. The pigment showed high absorbance in the UV region and decreased absorbance when shifted towards the visible region. The melanin pigment was further charecterized by FT-IR and EPR spectroscopy. A key enzyme 4-hydroxyphenylacetic acid hydroxylase catalyzes the formation of melanin pigment by hydroxylation of L-tyrosine was detected in this bacterium. Inhibition studies with specific inhibitor kojic acid and KCN proved that melanin is synthesized by DOPA ? Melanin pathway.
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KEYWORD
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melanin, pigment, spectroscopy, L-tyrosine, Klebsiella sp., 4-hydroxyphenylacetate hydroxylase
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