Abstract

ISOLATION AND CHARACTERIZATION OF TYROSINASE PRODUCING STREPTOMYCES LUTEOGRISEUS.

Gare Sandip Subhash and Kulkarni S.W.*

Abstract

The majority of actinomycetes are free living, saprophytic bacteria found widely distributed in soil, water and colonizing plants. Several species of Streptomyces genus produces bioactive molecules like antibiotics, pigments and many extracellular enzymes as glucose isomerase, amylase, cellulases and proteases. The present study was focused on isolation and characterization of Streptomyces producing tyrosinase enzyme. A total four soil samples were collected from the Shirala region Dist-Sangli. The total 70 actinomycetes isolates were obtained from soil by performing serial dilution technique and using Glycerol asparagine agar supplemented with Cycloheximide (100μg/ml). The primary screening of 70 isolates for tyrosinase production was carried out on skimmed milk agar. 19 isolates showed zone of hydrolysis and these were taken for secondary screening by using tyrosine agar medium and peptone yeast extract iron agar. The blackish brown diffusible pigment was reported on tyrosine agar and peptone yeast extract iron agar. The qualitative test for tyrosinase activity were carried out by inoculating in tyrosine broth supplemented with traces of chloroform and incubating at 300 C for 48 hrs. Red color was reported in tyrosine broth indicated positive tyrosinase activity. Among 19 isolates, 5 were reported as potential tyrosinase producers on the basis of intensity of red color in tyrosine broth. Out of them isolate C7 was identified on the basis of morphological, cultural, biochemical and 16S rRNA gene sequencing as Streptomyces luteogriseus.

Keywords: Actinomycetes, Streptomyces, Tyrosinase, Screening.