Abdel Malak C A, Abou El Adab I F, Vukashinovic V, Zalunin I A, Timokhina E A, Lavrenova G I, Stepanov V M
Chemistry Department, Mansoura University, Egypt.
Comp Biochem Physiol B Biochem Mol Biol. 1996 Jan;113(1):57-62. doi: 10.1016/0305-0491(96)02041-x.
Buffalo chymosin was isolated from abomasum mucosa extract of buffalo calves by affinity chromatography on gramicidin S-agarose followed by ion exchange chromatography on gamma-aminopropylsilochrom. Its molecular weight, 36 +/- 1 kDa, is similar to that of bovine calf chymosin. The N-terminal sequence Gly-Glu-Val-Ala-Ser-Val-Pro- coincides with that of bovine enzyme, whereas some differences were found in the amino acid composition of these enzymes. Buffalo and bovine enzyme possess similar but not identical structures. General proteolytic and milk-clotting activities of buffalo chymosin are also similar to those of bovine proteinase. pH-Optimum of its activity against hemoglobin lies at pH 4.0, somewhat higher than that for bovine chymosin, which indicates subtle differences in the functional properties of two enzymes.
通过在短杆菌肽S -琼脂糖上进行亲和层析，随后在γ-氨丙基硅基硅胶上进行离子交换层析，从水牛犊皱胃黏膜提取物中分离出水牛凝乳酶。其分子量为36±1 kDa，与牛犊凝乳酶相似。N端序列Gly-Glu-Val-Ala-Ser-Val-Pro-与牛凝乳酶的序列一致，然而在这些酶的氨基酸组成上发现了一些差异。水牛和牛凝乳酶具有相似但不完全相同的结构。水牛凝乳酶的一般蛋白水解活性和凝乳活性也与牛蛋白酶相似。其对血红蛋白的活性最适pH值为4.0，略高于牛凝乳酶，这表明两种酶的功能特性存在细微差异。
