Recombinant expression and characterization of N-acetylglucosaminyltransferase I derived from Nicotiana tabacum.
The C-terminal catalytic domain of tobacco N-acetylglucosaminyltransferase I fused to maltose-binding protein was produced in Escherichia coli as a soluble form with significant activity. The protein was affinity-purified using amylose resin, and its enzymatic properties were investigated, including its divalent cation requirements, optimal temperature, optimal pH, and substrate specificity.
Dohi
Koji
K
International Center for Biotechnology, Osaka University, 2-1 Yamada-oka, Suita, Osaka 565-0871, Japan.
Isoyama-Tanaka
Junko
J
Tokuda
Toru
T
Fujiyama
Kazuhito
K
D001483:Base Sequence; D001709:Biotechnology; D002240:Carbohydrate Sequence; D020134:Catalytic Domain; D017931:DNA Primers / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D033902:Periplasmic Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D014026:Nicotiana / Q000201:enzymology* / Q000235:genetics
D017931:DNA Primers; D018744:DNA, Plant; D029968:Escherichia coli Proteins; C461619:MalE protein, E coli; D009844:Oligosaccharides; D033902:Periplasmic Binding Proteins; D011993:Recombinant Fusion Proteins; D017351:N-Acetylglucosaminyltransferases; C024850:alpha-1,3-mannosyl-glycoprotein beta-1,2-N-acetylglucosaminyltransferase I