以凡纳滨对虾 (Litopenaeus vannamei shrimp) 虾头为原料，采用Q- Sepharose F F和Sephadex G-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Vector NTI 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.
Abstract
Endogenous alkaline protease from the head of Litopenaeus vannamei shrimp was purified by Q-Sepharose FF and SephadexG-150 chromatography. SDS-PAGE showed a single band of 79.95 kD. Endogenous acidic protease was purified by DEAESepharose FF and SephadexG100 chromatography.And SDSPAGE showed a single band at molecular weight of 27.45 kD. The animo acid sequences were detected by reversedphase highperformance liquid chromatography with electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS). The trypsin and pepsin digestion profiles were identified through matching NCBI database. The sequence alignment of the two proteases with proteases obtained from other species was performed using Vector NTI suite 8.0. A LSSPATLVSRVATVSLPR segment in the alkaline protease was found to be homologous to trypsin from pig. A terminal EFGLSETEPGTNF segment of the acidic protease was homologous to gastricsin from Xenopus laevis.
关键词
凡纳滨对虾虾头 /
内源碱性蛋白酶 /
内源酸性蛋白酶 /
胰蛋白酶 /
胃亚蛋白酶 /
同源性
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Key words
head of Litopenaeus vannamei shrimp /
endogenous alkaline protease endogenous acidic protease trypsin pepsin homology
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中图分类号：
Q78
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