Comparing substrate specificity of two UDP-sugar pyrophosphorylases and efficient one-pot enzymatic synthesis of UDP-GlcA and UDP-GalA.
Uridine 5'-diphosphate-glucuronic acid (UDP-GlcA) and UDP-galacturonic acid (UDP-GalA), the unique carboxylic acid-formed sugar nucleotides, are key precursors involved in the biosynthesis of numerous cell components. Limited availability of those components has been hindering the development of efficient ways towards facile synthesis of bioactive glycans such as glycosaminoglycans. In current study, we biochemically characterized two UDP-sugar pyrophosphorylases from Arabidopsis thaliana (AtUSP) and Bifidobacterium infantis ATCC15697 (BiUSP), and compared their activities towards a panel of sugar-1-phosphates and derivatives. Both enzymes showed significant pyrophosphorylation activities towards GlcA-1-phosphate, and AtUSP also exhibited comparable activity towards GalA-1-phosphate. By combining with monosaccharide-1-phosphate kinases, we have developed an efficient and facile one-pot three-enzyme approach to quickly obtain hundreds milligrams of UDP-GlcA and UDP-GalA.
Department of Chemistry and Center for Diagnostics & Therapeutics, Georgia State University, Atlanta, GA 30303, USA.
Yuxi
Y
Guo
National Glycoengineering Research Center, Shandong University, Jinan, Shandong 250100, China.
Junqiang
J
Fang
Department of Chemistry and Center for Diagnostics & Therapeutics, Georgia State University, Atlanta, GA 30303, USA.
Tiehai
T
Li
Department of Chemistry and Center for Diagnostics & Therapeutics, Georgia State University, Atlanta, GA 30303, USA.
Xu
X
Li
Department of Chemistry and Center for Diagnostics & Therapeutics, Georgia State University, Atlanta, GA 30303, USA.
Cheng
C
Ma
Department of Chemistry and Center for Diagnostics & Therapeutics, Georgia State University, Atlanta, GA 30303, USA.
Xuan
X
Wang
Department of Chemistry and Center for Diagnostics & Therapeutics, Georgia State University, Atlanta, GA 30303, USA; National Glycoengineering Research Center, Shandong University, Jinan, Shandong 250100, China. Electronic address: pwang11@gsu.edu.
Peng G
PG
Wang
Department of Chemistry and Center for Diagnostics & Therapeutics, Georgia State University, Atlanta, GA 30303, USA. Electronic address: lli22@gsu.edu.
Lei
L
Li
D017360:Arabidopsis / Q000201:enzymology; D029681:Arabidopsis Proteins / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D001644:Bifidobacterium / Q000201:enzymology; D055162:Biocatalysis; D007700:Kinetics; D009713:Nucleotidyltransferases / Q000737:chemistry*; D013379:Substrate Specificity; D014535:Uridine Diphosphate Glucuronic Acid / Q000737:chemistry*; D014539:Uridine Diphosphate Sugars / Q000737:chemistry*
D029681:Arabidopsis Proteins; D001426:Bacterial Proteins; D014539:Uridine Diphosphate Sugars; D014535:Uridine Diphosphate Glucuronic Acid; C050306:UDP-galacturonic acid; D009713:Nucleotidyltransferases; C514559:UDP-sugar pyrophosphorylase, Arabidopsis
Synthesis; UDP-GalA; UDP-GlcA; UDP-sugar pyrophosphorylase