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以凡纳滨对虾(Litopenaeus vannamei shrimp)虾头为原料,采用Q-Sepharose F F和SephadexG-150对虾头内源碱性蛋白酶进行了纯化,通过SDS-PAGE测定分子量为79.95 kD;采用DEAE-Sepharose F.F和Sephadex G-100对内源酸性蛋白酶进行了纯化,通过SDS-PAGE测定分子量为27.45 kD.利用HPLC-ESI-MS/MS对虾头内源碱性和酸性蛋白酶同源性进行了初步分析,将检测到内源性蛋白酶的部分氨基酸序列分别与不同物种的胰蛋白酶和胃蛋白酶氨基酸序列于VectorNTI suite 8.0软件上进行序列比对.结果表明,内源性碱性蛋白酶与猪胰蛋白酶具有很高的同源性,均含有氨基酸序列LSSPATLNSRVATVSLPR;内源性酸性蛋白酶与非洲蟾蜍胃亚蛋白酶具有很高的同源性,均含有氨基酸序列EFGLSETEPGTNF.
The shrimp head of Litopenaeus vannamei shrimp was used as raw material to purify shrimp head endogenous alkaline protease with Q-Sepharose FF and Sephadex G-150. The molecular weight was 79.95 kD by SDS-PAGE. And Sephadex G-100, the molecular weight of which was 27.45 kD by SDS-PAGE.HPLC-ESI-MS / MS was used to analyze the endogenous basic and acidic protease homology in shrimp head, A partial amino acid sequence of the endogenous protease was detected by sequence alignment with the trypsin and pepsin amino acid sequences of different species on a VectorNTI suite 8.0 software.The results showed that endogenous alkaline protease and porcine trypsin have high Homology with the amino acid sequence LSSPATLNSRVATVSLPR. The endogenous acidic protease has high homology with African toad gastroprotein, and contains the amino acid sequence EFGLSETEPGTNF.