INVESTIGATION OF KERATINASE ACTIVITY BY THERMO-ALKANOPHILIC Nocardiopsis sp. SD6 ISOLATED FROM FEATHER WASTE SOIL
Keywords:
Feather Waste, Keratinase, Nocardiopsis sp., ZymogramAbstract
The aim of the study was to isolate keratinolytic actinobacteria from feather waste soil,
poultry farm soil in Tiruchirappalli and Nammakkal District, Tamil Nadu, India and
their keratinase activity were investigated by feather degradation. Totally 91 isolates
were obtained from feather waste soil and poultry waste soil samples. Among the 91
isolates, 32 positive isolates were selected after grown on modified starch casein agar
(SCA) medium with additional casein. All 32 proteolytic isolates were subjected for
casein hydrolysis and four significant isolates were selected based on their growth on
basal liquid medium containing chicken feather. The isolate SD6 was tentatively
identified one of the best among 4 isolates. The isolate SD6 was characterized by
morphological, biochemical and molecular property inducing 16s rRNA analysis based
and has been identified as Nocardiopsis sp. SD6 (JF907189). The isolate was grown on
modified starch casein broth (SCB). Supernatant from centrifuged culture was examined
for protease and keratinase activity. The optimum temperature and pH for enzyme
activity for both proteolytic and keratinolytic was found at 50°C, pH 8 respectively.
Extracellular keratinase enzyme was separated from the culture supernatant by
centrifugation and precipitated with ammonium sulphate (80% saturation) and followed
by dialysis using dialysis membrane 150. Native PAGE was performed with the enzyme
sample and visible bands were observed after de-staining. Zymogram gel electrophoresis
was employed to identify the keratinase enzymes band and three bands in the gel were
found with positive keratinase activity.
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