NDLI: Characterization of uridine diphosphate-sugar pyrophosphorylase from Populus deltoids

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Characterization of uridine diphosphate-sugar pyrophosphorylase from Populus deltoids

Content Provider	Springer Nature Link
Author	Kim, Bong Gyu Ahn, Joong Hoon
Copyright Year	2013
Abstract	Nucleotide sugars serve as sugar donors for the biosynthesis of various cell components including cell wall, glycoproteins, and small molecules. Among them, uridine diphosphate (UDP)-glucose is one of main nucleotide sugars that serve as a substrate for the synthesis of other UDP-sugars. UDP-sugar pyrophosphorylase (USPase) mediates the formation of UDP-glucose from uridine triphosphate (UTP) and glucose-1-phosphate. A USPase, PdUSPase was cloned from Populus deltoids and expressed in Escherichia coli as glutathione Stransferase fusion protein. The purified recombinant PdUSPase catalyzed the reaction for the formation of UDP-glucose from glucose-1-phosphate and UTP, and for the formation of UDP-galactose from galactose-1-phosphate and UTP. However, the enzyme did not show any activity toward mannose-1-phosphate and UTP. These results indicate that PdUSPase belonging in UGPase A in phylogenetic analysis is the first UDP-glucose synthesizing enzyme showing a discrepancy between phylogenetic analysis and substrate range. E. coli complementation was also carried out to confirm the function of PdUSPase using E. coli galU mutant, which was mutated in UTP glucose-1-phosphate uridyltransferase. The galU mutant was transformed with the PdUGTase gene and a flavonoid glucosyl-transferase gene, AtUGT78D2. The resulting transformant was able to convert quercetin into quercetin 3-O-glucose similarly to that by the wild type E. coli strain harboring AtUGT78D2. These results indicated that PdUSPase catalyzed the formation of UDP-glucose from UTP and glucose-1-phosphate.
Starting Page	525
Ending Page	531
Page Count	7
File Format	PDF
ISSN	17382203
Journal	Journal of the Korean Society for Applied Biological Chemistry
Volume Number	56
Issue Number	5
e-ISSN	2234344X
Language	English
Publisher	Springer Netherlands
Publisher Date	2013-10-31
Publisher Place	Dordrecht
Access Restriction	Subscribed
Subject Keyword	Escherichia coli UTP glucose 1-phosphate uridylyltransferase mutant uridine diphosphate-sugar pyrophosphorylase uridine diphosphate-glucose Applied Microbiology Biological Techniques Bioorganic Chemistry
Content Type	Text
Resource Type	Article
Subject	Organic Chemistry Biochemistry, Genetics and Molecular Biology

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Characterization of uridine diphosphate-sugar pyrophosphorylase from Populus deltoids

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