Studies on the substrate specificity of Escherichia coli galactokinase.
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]
Yang
Jie
J
Laboratory for Biosynthetic Chemistry, Pharmaceutical Sciences Division, School of Pharmacy, University of Wisconsin-Madison, 777 Highland Avenue, Madison, WI 53705, USA.
Fu
Xun
X
Jia
Qiang
Q
Shen
Jie
J
Biggins
John B
JB
Jiang
Jiqing
J
Zhao
Jingjing
J
Schmidt
Joshua J
JJ
Wang
Peng G
PG
Thorson
Jon S
JS
D003124:Colorimetry; D004926:Escherichia coli / Q000201:enzymology*; D005686:Galactokinase / Q000737:chemistry* / Q000378:metabolism*; D005690:Galactose / Q000031:analogs & derivatives / Q000378:metabolism; D006027:Glycosides / Q000737:chemistry; D007700:Kinetics; D012459:Salicylates / Q000737:chemistry; D013379:Substrate Specificity
D006027:Glycosides; D012459:Salicylates; D005686:Galactokinase; D005690:Galactose