The invention provides a method for screening a I type 17 beta-
hydroxysteroid dehydrogenase inhibitor through an
immobilized enzyme, and belongs to the technical field of enzymology and
enzyme engineering. I type 17 beta-
hydroxysteroid dehydrogenase (17 beta-HSD1) has the important function in treating
hormone-dependent diseases. At present, a substrate radiolabelling method is mainly adopted in studying the activity of 17 beta-HSD1, due to the fact that the free
enzyme of 17 beta-HSD1 is likely to be inactivated, 17 beta-HSD1 is hard to prepare, a fresh
placenta is needed for preparing an
enzyme source in a new
medicine screening experiment of every time, raw materials are hard to obtain, the price is high, and difficulty is brought to the screening work of new
medicine. According to the method, an amino-modified
silicon ball is adopted as a carrier, a
glutaraldehyde crosslinking method is utilized, 17 beta-HSD1 extracted from the
placenta is fixed, an external immobilization 17 beta-HSD1 enzyme model is built,
androstenedione is adopted as a substrate, a
high performance liquid chromatography method is used for detecting products, and the potential 17 beta-HSD1 inhibitor is screened. According to the method,
instability of the free enzyme is overcome, operation is easy, the manufacturing cost is low, and repeated using is achieved.