Enzymatic characterization and comparison of various poaceae UDP-GlcA 4-epimerase isoforms.
UDP-alpha-D-galacturonic acid (UDP-GalA) is a key precursor for the synthesis of various bacterial and plant polysaccharides. UDP-glucuronic acid 4-epimerase (UGlcAE) catalyses the reversible conversion of UDP-alpha-D-glucuronic acid to UDP-GalA. UGlcAEs isolated from bacterial species have different biochemical properties when compared with the isoenzymes from the plant dicot species, Arabidopsis. However, little is known about the specificity of UGlcAE in Poaceae species. Therefore, we cloned and expressed in Escherichia coli several maize and rice UGlcAE genes, and compared their enzymatic properties with dicot homologs from Arabidopsis. Our data show that UGlcAE isoforms in different plant species have different enzymatic properties. For example, the Poaceae UGlcAE enzymes from rice and maize have significantly lower K(i) for UDP-xylose when compared with the Arabidopsis enzymes. The epimerases from different plant species are very specific and unlike their bacterial homolog in Klebsiella pneumoniae, can only use UDP-GlcA or UDP-GalA as their substrate. This study demonstrates that although members of the plant UGlcAE isoforms are highly conserved, the in vitro enzymatic activity of specific Poaceae isoform(s) may be regulated differently by specific nucleotide or nucleotide sugar.
Gu
Xiaogang
X
Department of Plant Biology, University of Georgia, Athens, GA 30602-4712, USA.
Wages
Christopher J
CJ
Davis
Kathryn E
KE
Guyett
Paul J
PJ
Bar-Peled
Maor
M
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology; D002238:Carbohydrate Epimerases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D003001:Cloning, Molecular; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D012275:Oryza / Q000201:enzymology*; D013045:Species Specificity; D013997:Time Factors; D003313:Zea mays / Q000201:enzymology*
D007527:Isoenzymes; D002238:Carbohydrate Epimerases