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OALib Journal期刊
ISSN: 2333-9721
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基于黄单胞菌冰核蛋白n端的表面展示体系建立
DOI: 10.3724/SP.J.1145.2014.10001, PP. 351-356
Keywords: 野油菜黄单胞菌,冰核蛋白,细菌表面展示,脂肪酶,大肠杆菌
Abstract:
为了建立一个自主、高效的细菌表面展示体系，利用野油菜黄单胞菌冰核蛋白n端结构域，将短小芽孢杆菌脂肪酶展示于大肠杆菌rosetta2(de3)表面.采用细胞分级、脂肪酶活测定和his-tagin-gelstain分析等检测了重组蛋白的表达情况，并进一步检测了不同底物、温度、ph和有机溶剂对酶活稳定性的影响.结果显示融合蛋白在大肠杆菌细胞外膜成功展示，重组菌脂肪酶活为(74.6±4.5)u/g冻干细胞，展示水平为每个细胞表面19267个脂肪酶分子；展示酶对中长链脂肪酸酯表现出较高的水解活性，最适ph为11.0，最适温度为40℃，且在40℃保温一周仍保持80%以上的酶活.本研究表明基于野油菜黄单胞菌冰核蛋白n端的展示体系是一种高效、稳定的细菌表面展示体系
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