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KMID : 0903520080510030153
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Journal of the Korean Society of Agricultural Chemistry and Biotechnology
2008 Volume.51 No. 3 p.153 ~ p.158
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Purification and Characteristics of Tyrosinase Inhibitor Produced by Actinomycetes F-97
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Bang Byung-Ho
Rhee Moon-Soo
Kim Jin-O
Yi Dong-Heui
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Abstract
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An actinomycetes F-97 producing tyrosinase inhibitor was isolated from soil samples. Isolation and purification of tyrosinase inhibitor produced by F-97 was performed as follows: IRC-120 (NH4+ type) column chromatography, silica gel column chromatography, C18 column chromatography and Sephadex LH-20 column chromatography were used successively after the centrifuged supernatant was adjusted to pH 4.0. To identify the purity of the inhibitor, octadecylsilyl(ODS) HPLC was carried out with 5% methanol as a mobile phase. Finally, the purification yield of a tyrosinase inhibitor was 5.24%. The inhibitor was very soluble in water, methanol and ethanol but insoluble in acetone, butanol, ethylacetate and chloroform. The λmax value of this inhibitor in water was 194 nm under UV light. The biochemical test of the inhibitor was positive in Molish, Benedict, conc. H2SO4, and KMnO4 tests but negative in iodine, ninhydrin, Million, Sakaguchi, xanthoproteic and Emerson tests. The tyrosinase inhibitor was stable against heat treatment of 100oC for 50 minutes and pH 4~9. The IC50 value of this inhibitor was 19.2 μg/ml for mushroom tyrosinase. In 1,000 μg/ ml inhibitor concentration, inhibition zone was 27 mm for Streptomyces bikiniensis NRRL B-1049. The inhibition of F-97 against mushroom tyrosinase was competitive with tyrosine.
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KEYWORD
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Actinomycetes, inhibitor, melanin, tyrosinase
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