title: Studies on the substrate specificity of Escherichia coli galactokinase.
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]
authors.affiliation: Laboratory for Biosynthetic Chemistry, Pharmaceutical Sciences Division, School of Pharmacy, University of Wisconsin-Madison, 777 Highland Avenue, Madison, WI 53705, USA.
authors.forename: Jie
authors.initials: J
authors.lastname: Yang
authors.forename: Xun
authors.initials: X
authors.lastname: Fu
authors.forename: Qiang
authors.initials: Q
authors.lastname: Jia
authors.forename: Jie
authors.initials: J
authors.lastname: Shen
authors.forename: John B
authors.initials: JB
authors.lastname: Biggins
authors.forename: Jiqing
authors.initials: J
authors.lastname: Jiang
authors.forename: Jingjing
authors.initials: J
authors.lastname: Zhao
authors.forename: Joshua J
authors.initials: JJ
authors.lastname: Schmidt
authors.forename: Peng G
authors.initials: PG
authors.lastname: Wang
authors.forename: Jon S
authors.initials: JS
authors.lastname: Thorson
mesh_terms: 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
chemical_list: D006027:Glycosides; D012459:Salicylates; D005686:Galactokinase; D005690:Galactose