Rojanavanich V, Yoshiike T, Tsuboi R, Takamori K, Ogawa H
Department of Dermatology, Juntendo University School of Medicine, Tokyo, Japan.
Infect Immun. 1990 Sep;58(9):2856-61. doi: 10.1128/iai.58.9.2856-2861.1990.
An extracellular proteinase from a fast-growing strain of Hendersonula toruloidea was demonstrated when it was cultivated in liquid medium containing bovine serum albumin as the sole nitrogen source. Purification to homogeneity of the proteinase was performed by carboxymethyl cellulose, CM Sephadex G-50, and Sephacryl S-200 column chromatographies. The purified enzyme was a chymotrypsinlike serine proteinase, as indicated by the strong inhibitory activities of diisopropyl fluorophosphate, phenylmethylsulfonyl fluoride, L-1-tosylamido-2-phenylethylchloromethyl ketone, and chymostatin and good kinetic constants for a synthetic substrate, Suc-Ala-Ala-Pro-Phe-MCA. The enzyme had a pI of 8.4, a pH optimum of 9.0, and a molecular weight of 34,000. Skin constituents such as stratum corneum and nail, but not hair, were easily digested by this enzyme. Thus, this extracellular proteinase may play a role in the invasion of thickly keratinized skin and nail by this organism.
当亨德逊酵母快速生长菌株在以牛血清白蛋白作为唯一氮源的液体培养基中培养时，可证明其能产生一种胞外蛋白酶。通过羧甲基纤维素、CM Sephadex G - 50和Sephacryl S - 200柱色谱法将该蛋白酶纯化至同质。纯化后的酶是一种类胰凝乳蛋白酶丝氨酸蛋白酶，这可通过二异丙基氟磷酸酯、苯甲基磺酰氟、L - 1 - 甲苯磺酰氨基 - 2 - 苯乙基氯甲基酮和抑肽酶的强抑制活性以及对合成底物Suc - Ala - Ala - Pro - Phe - MCA的良好动力学常数来表明。该酶的pI为8.4，最适pH为9.0，分子量为34000。角质层和指甲等皮肤成分，而非毛发，很容易被这种酶消化。因此，这种胞外蛋白酶可能在该生物体侵入角质化程度高的皮肤和指甲过程中发挥作用。
