重组L-天冬氨酸-β-脱羧酶融合蛋白复性条件的研究

Study on Renaturation Conditions of Recombinant L-Aspartate-β-Decarboxylase

以本实验室构建保存的天冬氨酸脱羧酶工程菌为研究材料,通过基因表达,对所得到的重组胞内融合表达型天冬氨酸脱羧酶蛋白进行体外复性研究,考察温度、复性液的pH、复性时间、复性液种类等因素对重组天冬氨酸脱羧酶体外复性的影响。结果表明:最佳复性液为pH8.7的Tris-HCl缓冲液,在保温时间2h,温度40℃下,复性效果最好。另外一些pH能达到8.7的缓冲液复性效果都不如Tris-HCl缓冲液。

Recombinant L-aspartate-β-decarboxylase expressed by reconstructed engineering bacteria was purified and investigated on refolding conditions.The results indicated that the enzyme showed highest activity when treated with Tris-HCl buffer of pH 8.7 and incubated at 40 ℃ for 2 hours.