Characterization of an alkaline active-thiol forming extracellular serine keratinase by the newly isolated Bacillus pumilus.
OBJECTIVE
The aim of the study was to optimize microbial degradation of keratinous waste and to characterize the alkaline active keratinase showing its biotechnological importance.


RESULTS
An extracellular keratinase enzyme was purified from the culture medium of a bacterial isolate and the conditions were optimized. The molecular weight of DEAE-Sepharose-purified keratinase was determined by SDS-PAGE. Instrumental analyses were investigated to study the mechanism of bovine hair hydrolysis. Isolate was identified as Bacillus pumilus based on phenotypic characteristics and 16S rDNA sequence. The optimized condition for its growth was pH 8 and 35 degrees C. The molecular weight of the keratinase was estimated as 65 kDa. Activity inhibition by phenyl methyl sulphonyl fluoride confirmed keratinase as serine protease type. Instrumental analysis revealed the sulphitolysis and proteolysis involved mechanism in bovine hair hydrolysis.


CONCLUSIONS
This study indicates that the isolated keratinase is an alkaline active serine protease with a high degree of activity towards bovine hair.


CONCLUSIONS
This study examines a serine protease with high keratinolytic activity and degradation mechanism for bovine hair. The keratinolytic activity of the isolated strain and its reaction mechanism on bovine hair could show biotechnological potential in the leather industry.
Kumar
A Ganesh
AG
Department of Environmental Technology, Central Leather Research Institute, Chennai, India.
Swarnalatha
S
S
Gayathri
S
S
Nagesh
N
N
Sekaran
G
G
D000818:Animals; D001407:Bacillus / Q000302:isolation & purification / Q000378:metabolism*; D001673:Biodegradation, Environmental; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D005285:Fermentation; D006197:Hair; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007218:Industrial Microbiology; D007633:Keratins; D008855:Microscopy, Electron, Scanning; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000302:isolation & purification*; D012697:Serine Endopeptidases / Q000302:isolation & purification; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013633:Tanning; D013696:Temperature
D013438:Sulfhydryl Compounds; D007633:Keratins; D010447:Peptide Hydrolases; C035855:keratinase; D012697:Serine Endopeptidases