β3GnT8/β3GalT7催化活性的研究

Research of the Catalytic Activity of β3GnT8/β3GalT7

目的进一步验证β1,3半乳糖基转移酶(β3GalT7)的催化功能。方法将重组表达载体pGEX-6p-1-β3GnT8/GalT7转化BL21大肠杆菌,经IPTG诱导表达后,得到GST-β3GnT8/GalT7融合蛋白,分别进行β3GalT7和β3GnT8反应体系的酶促反应,利用高效液相色谱(HPLC)进行检测。结果HPLC检测结果表明,β3GalT7同时具有β3GalT7和β3GnT8的活性。结论β3GalT7具有双重酶活性,拟初步重命名为β3GnT8/β3GalT7。

Objective To determine the catalytic activity of the β1,3-N-acetylglucosyltransferase （β3GnT8）/β1,3-galactosyltransferases （β3GalT7）. Methods The recombinant plasmid was transformed into E. Coli BL21. Then a fusion protein was expressed after the induction by IPTG. After testified by Western blot,we purified the expression products by affinity chromatography. After renaturation,its enzyme activity was assayed by HPLC. Results It suggested that β3GalT7 own the catalytic activities of β3GalT7 and β3GnT8. Conclusion It has both activities of GalTs and GnTs,so we renamed it as β3GnT8/β3GalT7.