Azarenkova N M, Vaganova T I, Strongin A Ia, Stepanov V M
Biokhimiia. 1976 Jan;41(1):20-6.
Caboxypeptidase with pH optimum 4--5 for peptide substrates is isolated from "oryzine", the enzyme mixture produced by Aspergillus oryzae, by means of successive salt fractionation, Sephadex G-75 gel filtration, chromatography on amberlite IRC-50, hydroxylapatite, DEAE-cellulose and polyacrylamide gel electrophoresis. Its molecular weight, as determined by means of polyacrylamide gel electrophoresis in the presence of sodium dolecylsulphate, was found to be 37 000. The enzyme has a broad substrate specificity and does not possess dipeptidase and esterase activities. Acid carboxypeptidase from Asp. oryzae is not a metalloenzyme, it is inactivated by specific inhibitors of serine proteases and by compounds blocking SH-groups. The enzyme is suggested to contain functionally important serine and cysteine residues and to be acid carboxipeptidase.
通过连续盐分级分离、Sephadex G - 75凝胶过滤、在amberlite IRC - 50、羟基磷灰石、DEAE - 纤维素上的色谱法以及聚丙烯酰胺凝胶电泳，从米曲霉产生的酶混合物“oryzine”中分离出对肽底物而言最适pH为4 - 5的羧肽酶。通过在十二烷基硫酸钠存在下进行聚丙烯酰胺凝胶电泳测定，其分子量为37000。该酶具有广泛的底物特异性，且不具有二肽酶和酯酶活性。米曲霉的酸性羧肽酶不是金属酶，它可被丝氨酸蛋白酶的特异性抑制剂以及阻断SH基团的化合物所灭活。该酶被认为含有功能上重要的丝氨酸和半胱氨酸残基，并且是酸性羧肽酶。
