Black Fungus Ribosome Inactivating Protein Separation And Purification, And Nature Study

Ribosome-inactivating protein (RIP) are a group of toxic protein widely distributedin the nature, which has particular enzymology mechanism and biological functions.Although recently reports about RIP mostly come from higher plants, it was also foundin edible fungi. This experiment combining multi-protein purified method andtechnology successfully separated RIP from mycelium of Auriculariaauricular(Jew's-ear), in addition to identify their activity and quality, anion exchange onDE-52 cellulose, gel filtration on Sephadex G-75 and Aft-gel blue were used to purifiedfor RIP, the purified product was detected for RIP content,molecular weight andrestrained activity of protein synthesized in vitro by ultraviolet absorption spectroscopyand SDS-PAGE electrophoresis etc. Then re-purified the product bearing restrainactivity until obtain higher pure Jew's-ear ribosome inactivating protein G2, degree ofpurity was 99.3% through high performance liquid chromatography. The protein G2show positive by RNA N-glycosidase activity and suppressed protein in vitrotranslated synthesis activity measuring. As a result the G2 is Jew's-ear ribosomeinactivating protein, which is a new discovering protein, a new resource for fungiprotein and a new way to develop and apply RIP. This project do forward foundationresearch for additional clone ribosome inactivating protein gene.