Partial Purification And Characterization Of Trypsin And Angiotensin-Converting Enzyme Inhibitor From The Skin Of Bombyx Mori

Bombyx mori belong to invertebrate, insecta, Lepioloptera, Bombycidae. When theyhappen to the various stimulus of physical~ chemical and biologic factors, a series ofimmune substances were occurred in hemolymph such as antibacterial proteins,antibacterial peptides,bacteriolysis enzyme and lectin to antagonize the invasion ofexogenous microorganisms. These immune substances were expected to become new drugswith antibacterial,antivirus and anticancer activities. In order to detect if these antibacterialsubstances or any components with clinical activities in the skin of Bombyx mori, in thispaper the trypsin inhibitors and angiotensin-converting enzyme inhibitor were detected andpartial purified from the skin of the Bombyx mori, the main contents includes:1 Partial purification and characterization of trypsin inhibitor from. the skin of Bombyx moriInthis paper, a trypsin inhibitor was attemped to purify from skin of BOMBYX MORIand studied its partial characterization.The optima reaction condition was fixed on 0.1mg/ml Trypsin和0.075％gelation bymeasuring the effects of concentration of trypsin and gelation on native-PAGE.The crude extraction of the sample was salted out with 70％ammonium sulfate,dialyzed to obtain the crude extract of trypsin inhibitor.The. kinds and amounts of trypsin inhibitors from crude extract and hot stabilities ofevery bands of ~eiectrophoresis were studied with gelatin-PAGE. There existed eight trypsininhibitor bands from crude~ extract, and gelatin-PAGE showed two bands in this fraction cantolerant higher hot stability, there is no effects towards their activities after boiled.Further purification of crude extraction was carried out with DEAE-52 celluloseion-exchange .chromatograph, the results of purification and inhibiting activities wasmeasured with gelatin-PAGE. After DEAE-52 cellulose ion-exchange chromatograph, the relative hot stability higher proteins were eluted with buffered contained 0.1 NaCl.2 Detection and partial purification of ACEI from the skins of the Bombyx mori In this paper, by using the two different substrate of ACE to measure the activities ofACEI, and vitro screening models of ACEI was established. By using the two establishedmethods above, the inhibiting activities of captopril which is the special inhibitor of ACEIwere measured, and the results were accordant with the two methods and also showedaccordant with the document reported.Crude ACEI from the crude extraction of skin of Bombyx mori were detected, afterfurther purification of the crude, extraction, we found that ACEI were concentrated inelution apex which was eluted using buffer complemented with 0.1 M NaCl.