以变性染色皮粉为底物的蛋白酶活力检测方法研究  

Proteolytic Activity Determination of Proteases with Denatured Hide Power Labeled with Active Dye as Substrate

作　　者：李成霞[1] 许冰斌[1] 王然然[1] 梁亮[1] 彭必雨[1] 

机构地区：[1]四川大学制革清洁技术国家工程实验室,四川成都610065

出　　处：《中国皮革》2012年第5期19-23,27,共6页China Leather

基　　金：教育部新世纪优秀人才支持计划项目(NCET-10-0598);国家科技支撑计划项目(2011BAC06B00)

摘　　要：以活性染料Remazol brilliant blue R(RBB)对皮粉进行染色,制备了在较宽的pH范围内稳定的热变性染色皮粉。研究了反应时间、底物和蛋白酶浓度等因素对酶催化染色皮粉水解速度的影响,建立了一种基于变性皮粉-RBB底物的蛋白酶活力检测方法。用建立的方法,对几种常用的制革蛋白酶进行酶活力测定,并与常用的基于酪素底物的福林法和LVU法进行对比,结果表明:对于大部分蛋白酶,2种底物检测结果有较大差异。A kind of hide powder labeled covalently with Remazol Brilliant Blue R （RBB） was prepared, which was a kind of hot - denatured stained collagen because it was boiled during preparing. It was stable and suitable for assaying protease activity in a rather broad pH range. The influences of some key factors, such as reaction times, concentrations of hide powder substrate and enzyme, on proteolytic reaction were investigated, and a sensitive method for determining the proteolytic activity of protease based on the hide powder- RBB substrate was established. Then the proteolytic activities of some proteases frequently used in tanning process were assayed with the established method, and the results were compared with that of Folin and LVU methods based on the casein substrate. The results show that, for most of proteases, the proteolytic activity results based on the two different substrates were quite different.

关 键 词：染色皮粉 底物 蛋白酶 酶活力 

分 类 号：TS51[轻工技术与工程—皮革化学与工程]