Hannaert V, Opperdoes F R, Michels P A
Research Unit for Tropical Diseases, International Institute of Cellular and Molecular Pathology, Brussels, Belgium.
Protein Expr Purif. 1995 Jun;6(3):244-50. doi: 10.1006/prep.1995.1031.
The Trypanosoma brucei and Trypanosoma cruzi glycosomal glyceraldehyde-3-phosphate dehydrogenases have been overexpressed in Escherichia coli using a T7 expression system. These enzymes have been highly purified by ammonium sulfate precipitation, followed by phenyl-Sepharose and phospho-ultrogel chromatography. From 1 liter of bacterial culture, we obtained 4.4 mg of T. brucei enzyme, with a specific activity of 147 units/mg, and 26.6 mg of T. cruzi enzyme, with a specific activity of 122 units/mg. Both proteins have a similar subunit mass of 38 kDa. Some physicochemical and kinetic properties have been determined and compared with those reported for the authentic T. brucei enzyme. The two enzymes appear to be very similar, except for the dependence of their activity on ionic strength.
布氏锥虫和克氏锥虫的糖酵解体甘油醛-3-磷酸脱氢酶已通过T7表达系统在大肠杆菌中过表达。这些酶已通过硫酸铵沉淀，随后进行苯基琼脂糖和磷酸超凝胶色谱法进行了高度纯化。从1升细菌培养物中，我们获得了4.4毫克布氏锥虫酶，比活性为147单位/毫克，以及26.6毫克克氏锥虫酶，比活性为122单位/毫克。两种蛋白质的亚基质量相似，均为38 kDa。已测定了一些物理化学和动力学性质，并与报道的天然布氏锥虫酶的性质进行了比较。除了它们的活性对离子强度的依赖性外，这两种酶似乎非常相似。
