Kudinov S A, Kolodzeĭskaia M V, Malen'kikh L V, Konoplich L A, Aiushin N B
Prikl Biokhim Mikrobiol. 1986 Jan-Feb;22(1):53-8.
A simple and convenient technique was developed for isolation of the proteolytic enzyme complexes from the whale (Balaenoptera) pancreas. The proposed techniques enables the proteolytic complexes to be obtained with the protein yield 2.6 times higher than the classical procedure. The proteolytic activity increased 3.2 times (casein as a substrate), esterase activities, 1.4 times (N-benzoyl-L-tyrosine methyl ester as a substrate) and 1.2 times (N-alpha-benzoyl-L-arginine ethyl ester as a substrate). Soybean and barley trypsin inhibitors and ovomycoid in free and immobilized state inhibit the esterase activities of the proteolytic complexes. An additional purification of the proteolytic complexes was carried out using the affinity sorbent Soybean trypsin inhibitor--Sepharose 4B. The molecular weight of the enzymes determined by means of PAAG electrophoresis was found to be 20 000-20 500. The hydrolysis of some synthetic substrates by the proteolytic enzyme complexes obtained according to the proposed techniques was being studied.
开发了一种简单便捷的技术，用于从鲸（须鲸属）胰腺中分离蛋白水解酶复合物。所提出的技术能够获得蛋白产量比传统方法高2.6倍的蛋白水解复合物。蛋白水解活性提高了3.2倍（以酪蛋白为底物），酯酶活性提高了1.4倍（以N-苯甲酰-L-酪氨酸甲酯为底物）和1.2倍（以N-α-苯甲酰-L-精氨酸乙酯为底物）。游离和固定化状态的大豆和大麦胰蛋白酶抑制剂以及卵类粘蛋白可抑制蛋白水解复合物的酯酶活性。使用亲和吸附剂大豆胰蛋白酶抑制剂-琼脂糖4B对蛋白水解复合物进行了进一步纯化。通过聚丙烯酰胺凝胶电泳测定的酶分子量为20000-20500。正在研究根据所提出的技术获得的蛋白水解酶复合物对一些合成底物的水解作用。
