Bjelland S, Volden G, Raa J
Institute of Fisheries, University of Tromsø, Norway.
Arch Dermatol Res. 1988;280(1):50-3. doi: 10.1007/BF00412689.
A simple assay measuring degradation of human epidermal keratin and bovine tendon collagen is presented. Insoluble protein substrate (30 mg) was incubated with 1 ml buffer and enzyme sample for 1 h at 37 degrees C, following addition of 1 ml distilled water and removal of the remaining substrate by filtration/centrifugation. The protein content was determined in the filtrate/supernatant by the Lowry method. Keratin was prepared as follows: Freeze-drying, homogenization in a mortar or beetling mill, extraction in 0.9% (w/v) NaCl followed by water and 100% ethanol, drying at 37 degrees C. The assay was tested with pig pepsin, bovine trypsin, and crude extract of fish stomach, demonstrating that these preparations are effective in degrading human epidermal keratin.
本文介绍了一种用于测量人表皮角蛋白和牛肌腱胶原蛋白降解的简单检测方法。将不溶性蛋白质底物（30毫克）与1毫升缓冲液和酶样品在37℃下孵育1小时，然后加入1毫升蒸馏水，并通过过滤/离心去除剩余的底物。用洛瑞法测定滤液/上清液中的蛋白质含量。角蛋白的制备方法如下：冷冻干燥，在研钵或捣碎机中匀浆，用0.9%（w/v）NaCl提取，然后用水和100%乙醇提取，在37℃下干燥。用猪胃蛋白酶、牛胰蛋白酶和鱼胃粗提取物对该检测方法进行了测试，结果表明这些制剂能够有效降解人表皮角蛋白。
