Berezin I V, Ugarova N N, Fel'dman D P
Biokhimiia. 1977 May;42(5):926-33.
A procedure for isolation of stabilized preparations of peroxidase, covalently bound with AH-Sepharose 4B by the enzyme carbohydrate residues, which had been previously oxidized by iodine acid, is proposed. The catalytic properties and thermostability of soluble oxidized and immobilized peroxidase has been studied and a comparison of these properties with those of the native enzyme has been made. It has been demonstrated that the oxidation of the carbohydrate residues dose not affect eh Km value for soluble oxidized peroxidase, but leads to a decrease in the kcat approximately two times for the reaction of peroxidase exidation of o-dianizidine. In case of immobilized peroxidase the Km value for H2O2 does not change, whereas the Km value for o-dianizidine is decreased approximately 20 times. It is demostrated that the chemical structure of the matrix surface and in case of the same matrix used--the degree and type of modification of the functional groups of protein, have a strong effect on the stability of the immobilized enzyme.
提出了一种通过酶的碳水化合物残基与经碘酸预先氧化的AH-琼脂糖4B共价结合来分离过氧化物酶稳定制剂的方法。研究了可溶性氧化过氧化物酶和固定化过氧化物酶的催化特性和热稳定性，并将这些特性与天然酶的特性进行了比较。结果表明，碳水化合物残基的氧化不影响可溶性氧化过氧化物酶的Km值，但对于邻联茴香胺的过氧化物酶氧化反应，kcat大约降低两倍。对于固定化过氧化物酶，H2O2的Km值不变，而邻联茴香胺的Km值大约降低20倍。结果表明，基质表面的化学结构以及在使用相同基质的情况下——蛋白质官能团的修饰程度和类型，对固定化酶的稳定性有很大影响。
