一种新阿片肽的分离纯化

报道了一种新阿片肽OP1的分离纯化。将重组毕赤酵母经适宜的生长和表达培养后,所得的发酵液经离心得无细胞上清液,上清液经超滤后过Sephadex G-10柱。将经Sephadex G-10柱所得具有阿片活性的粗组分用HPLC-MS分析,根据阿片肽N-端均有一个酪氨酸残基,且在肽链的第三或第四位上有一个芳香族氨基酸残基这一性质,依据分子量确定活性组分中可能存在的所有阿片肽,然后根据这些阿片肽的等电点,利用AKTA Purifier 100快速纯化系统的DEAE-阴离子交换纤维素柱将其进一步分离,活性组分再用Sephasil peptide C18反相高压液相柱分离得到活性组分OP1肽,鉴定纯度后测定其氨基酸组成。最后确定该肽的一级序列为YPFPGPIRYG,该阿片肽序列目前尚未见报道。

Separating and Purification of a New Opioid Peptide

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.