Keratinase of Doratomyces microsporus
Keratinase of Doratomyces microsporus
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DOI：
10.1007/s002530050008
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发表时间：
2000-02-01
影响因子：
5
通讯作者：
Friedrich, J
中科院分区：
文献类型：
--
作者：
Gradisar, H;Kern, S;Friedrich, J
真菌小孢子菌在含有用于酶合成的蛋白质诱导剂的培养基中进行深层需氧培养时产生细胞外角蛋白酶。使用疏水相互作用层析随后使用凝胶层析将角蛋白酶纯化至均质。分子量估计为 33 kDa（通过 SDS-PAGE 分析）或 30 kDa（通过凝胶色谱），表明具有单体结构。酶的等电点确定为9左右。角蛋白分解活性的最佳pH和温度分别为pH 8-9和50℃。丝氨酸蛋白酶抑制剂PMSF完全抑制角蛋白酶。该酶没有被糖基化。它能够水解不同的角蛋白材料以及一些非角蛋白。对已知蛋白酶特异的一些合成底物的水解表明，小孢子菌的角蛋白酶与蛋白酶 K 接近。
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.