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Studies on the substrate specificity of Escherichia coli galactokinase
Authors:Yang Jie  Fu Xun  Jia Qiang  Shen Jie  Biggins John B  Jiang Jiqing  Zhao Jingjing  Schmidt Joshua J  Wang Peng G  Thorson Jon S
Institution:Laboratory for Biosynthetic Chemistry, Pharmaceutical Sciences Division, School of Pharmacy, University of Wisconsin-Madison, 777 Highland Avenue, Madison, WI 53705, USA.
Abstract:In vitro glycorandomization (IVG) technology is dependent upon the ability to rapidly synthesize sugar phosphates. Compared with chemical synthesis, enzymatic (kinase) routes to sugar phosphates would be attractive for this application. This work focuses upon the development of a high-throughput colorimetric galactokinase (GalK) assay and its application toward probing the substrate specificity and kinetic parameters of Escherichia coli GalK. The demonstrated dinitrosalicylic assay should also be generally applicable to a variety of sugar-processing enzymes. reaction: see text]
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