Keratinase of Doratomyces microsporus.
The fungus Doratomyces microsporus produced an extracellular keratinase during submerged aerobic cultivation in a medium containing a protein inducer for enzyme synthesis. The keratinase was purified to homogeneity using hydrophobic interaction chromatography followed by gel chromatography. The molecular weight was estimated to be 33 kDa (from SDS-PAGE analysis) or 30 kDa (by gel chromatography), suggesting a monomeric structure. The isoelectric point of the enzyme was determined to be around 9. The optimal pH and temperature for keratinolytic activity were pH 8-9 and 50 degrees C, respectively. The serine protease inhibitor PMSF totally inhibited the keratinase. The enzyme was not glycosylated. It was capable of hydrolysing different keratinous materials as well as some non-keratinous proteins. Hydrolysis of some synthetic substrates, specific for known proteinases, suggested that the keratinase of D. microsporus is close to proteinase K.
Gradisar
H
H
National Institute of Chemistry, Ljubljana, Slovenia. helena.gradisar@ki.si
Kern
S
S
Friedrich
J
J
D000818:Animals; D002364:Caseins / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004817:Epidermis / Q000378:metabolism; D005658:Fungi / Q000254:growth & development / Q000378:metabolism*; D006454:Hemoglobins / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D013379:Substrate Specificity; D013552:Swine; D013696:Temperature
D002364:Caseins; D004791:Enzyme Inhibitors; D006454:Hemoglobins; D010664:Phenylmethylsulfonyl Fluoride; D010447:Peptide Hydrolases; C035855:keratinase