title: Characterization of hyperthermostable fructose-1,6-bisphosphatase from Thermococcus onnurineus NA1.
abstract: To understand the physiological functions of thermostable fructose-1,6-bisphosphatase (TNA1-Fbp) from Thermococcus onnurineus NA1, its recombinant enzyme was overexpressed in Escherichia coli, purified, and the enzymatic properties were characterized. The enzyme showed maximal activity for fructose-1,6-bisphosphate at 95°C and pH 8.0 with a half-life (t (1/2)) of about 8 h. TNA1-Fbp had broad substrate specificities for fructose-1,6-bisphosphate and its analogues including fructose-1-phosphate, glucose-1-phosphate, and phosphoenolpyruvate. In addition, its enzyme activity was increased five-fold by addition of 1 mM Mg(2+), while Li(+) did not enhance enzymatic activity. TNA1-Fbp activity was inhibited by ATP, ADP, and phosphoenolpyruvate, but AMP up to 100 mM did not have any effect. TNA1-Fbp is currently defined as a class V fructose-1,6-bisphosphatase (FBPase) because it is very similar to FBPase of Thermococcus kodakaraensis KOD1 based on sequence homology. However, this enzyme shows a different range of substrate specificities. These results suggest that TNA1-Fbp can establish new criterion for class V FBPases.
authors.affiliation: Division of Life Science, Korea Basic Science Institute, Daejeon, 305-806, Republic of Korea.
authors.forename: Yeol Gyun
authors.initials: YG
authors.lastname: Lee
authors.forename: Sung Gyun
authors.initials: SG
authors.lastname: Kang
authors.forename: Jung-Hyun
authors.initials: JH
authors.lastname: Lee
authors.forename: Seung Il
authors.initials: SI
authors.lastname: Kim
authors.forename: Young-Ho
authors.initials: YH
authors.lastname: Chung
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D003001:Cloning, Molecular; D020536:Enzyme Activators / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D006597:Fructose-Bisphosphatase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D005636:Fructosephosphates / Q000378:metabolism; D015870:Gene Expression; D005958:Glucosephosphates / Q000378:metabolism; D006207:Half-Life; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000378:metabolism; D010728:Phosphoenolpyruvate / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity; D019712:Thermococcus / Q000201:enzymology*
chemical_list: D020536:Enzyme Activators; D004791:Enzyme Inhibitors; D005636:Fructosephosphates; D005958:Glucosephosphates; D011994:Recombinant Proteins; C032284:fructose-1-phosphate; D000244:Adenosine Diphosphate; D010728:Phosphoenolpyruvate; D000255:Adenosine Triphosphate; C031590:glucose-1-phosphate; D006597:Fructose-Bisphosphatase; D008274:Magnesium