Gaĭda A V, Rudenskaia G N, Stepanov V M
Biokhimiia. 1981 Nov;46(11):2064-73.
Using affinity chromatography on bacitracin-Sepharose combined with ion-exchange chromatography on aminosilochrome and isoelectrofocusing, individual serine proteases have been isolated for the first time from surface cultures of T. lignorum and T. koningii. The proteinases have pI values at 6.8 and 6.7 and pH optima of 10.5. Both proteinases are stable within the pH range of 4-11 and have molecular weight of 21 000. The amino acid composition of T. lignorum enzyme is Lys3His4Arg9Asx23Thr17Ser23Glx10Pro8Gly27Ala25Cys3Val13Met2Ile11Leu11Tyr7Phe6Tr p3, that of the T. koningii enzyme is Lys3His4 Arg9Asx23Thr16Ser26Glx10Pro9Gly29Ala26Cys3Val14Met2Ile9Leu11Tyr6Phe5Trp3. The enzymes are completely inhibited by phenylmethylsulfonylfluoride, diphenylcarbamoylchloride and trypsin inhibitors from beans, Actinomyces janthinus and potato tubers. The enzymatic and molecular properties of the enzymes are similar to those of subtilisins and previously described fungal serine proteinases, especially to those of proteinase K of the fungus Tritirachium album Limber.
利用杆菌肽-琼脂糖亲和层析结合氨基硅铬上的离子交换层析和等电聚焦，首次从木霉和康宁木霉的表面培养物中分离出了单个丝氨酸蛋白酶。这些蛋白酶的等电点值分别为6.8和6.7，最适pH为10.5。两种蛋白酶在pH 4-11范围内都很稳定，分子量为21000。木霉酶的氨基酸组成为Lys3His4Arg9Asx23Thr17Ser23Glx10Pro8Gly27Ala25Cys3Val13Met2Ile11Leu11Tyr7Phe6Tr p3，康宁木霉酶的氨基酸组成为Lys3His4 Arg9Asx23Thr16Ser26Glx10Pro9Gly29Ala26Cys3Val14Met2Ile9Leu11Tyr6Phe5Trp3。这些酶被苯甲基磺酰氟、二苯基甲酰氯以及来自豆类、詹氏放线菌和马铃薯块茎的胰蛋白酶抑制剂完全抑制。这些酶的酶学和分子特性与枯草杆菌蛋白酶和先前描述的真菌丝氨酸蛋白酶相似，但与真菌白僵菌蛋白酶K的特性尤为相似。
