Separating and Purification of a New Opioid Peptide
一种新阿片肽的分离纯化

A new opioid peptide, OP1, was obtained by separating and purification. Supernatant was collected by centrifugation from the recombinant Pichia pastoris fermented. The supernatant was ultrafiltrated and then separated by Sephadex G-10 chromatography. On account of the common structural feature among endogenous and exogenous opioid peptides, that is, the presence of a tyrosine residue at the amino terminal end and the presence of another aromatic residue , e.g. phenylalanine or tyrosine, in the third or fourth position, all potential existent opioid peptides in zymotic supernatant can be obtained. According to the molecular weight of these opioid peptides identified by HPLC-MS, all possible opioid peptides in active component were ascertained. Depending on pI and the molecular weight of these opioid peptides, the active component was purified by the AKTA-100 Purifier system with DEAE-cellulose ion exchange chromatography and Sephasil peptide C18 reversed phase chromatography. The peptide OP1 was obtained finally. The OP1 showed one peak on capillary electrophoresis. The amino acid composition of OP1 was determined. The sequence of OP1 was YPFPGPIRYG.