Ngo T T, Lenhoff H M
Appl Biochem Biotechnol. 1983 Oct;8(5):407-14. doi: 10.1007/BF02779913.
A sensitive and rapid amperometric assay for collagenase has been developed. The substrate for the assay is glucose oxidase covalently linked to insoluble collagen with dimethylsuberimidate. The collagenase cleaves the insoluble collagen--glucose oxidase conjugate into smaller, soluble fragments that have glucose oxidase activity. That activity is proportional to the collagenase activity hydrolyzing the insoluble conjugate. In the absence of collagenase, no glucose oxidase activity is found in the soluble phase. Glucose oxidase activity was assayed by measuring amperometrically the rate at which hydrogen peroxide is produced. The kinetics follow that proposed for a soluble enzyme acting on an insoluble substrate.
已开发出一种灵敏且快速的胶原酶安培测定法。该测定法的底物是通过辛二酸二甲酯与不溶性胶原共价连接的葡萄糖氧化酶。胶原酶将不溶性胶原 - 葡萄糖氧化酶偶联物切割成具有葡萄糖氧化酶活性的较小可溶性片段。该活性与水解不溶性偶联物的胶原酶活性成正比。在没有胶原酶的情况下，在可溶相中未发现葡萄糖氧化酶活性。通过安培法测量过氧化氢产生的速率来测定葡萄糖氧化酶活性。动力学遵循作用于不溶性底物的可溶性酶的动力学。
