title: Production of glucose-6-phosphate by glucokinase coupled with an ATP regeneration system.
abstract: A process of glucose-6-phosphate (G-6-P) production coupled with an adenosine triphosphate (ATP) regeneration system was constructed that utilized acetyl phosphate (ACP) via acetate kinase (ACKase). The genes glk and ack from Escherichia coli K12 were amplified and cloned into pET-28a(+), then transformed into E. coli BL21 (DE3) and the recombinant strains were named pGLK and pACK respectively. Glucokinase (glkase) in pGLK and ACKase in pACK were both overexpressed in soluble form. G-6-P was efficiently produced from glucose and ACP using a very small amount of ATP. The conversion yield was greater than 97 % when the reaction solution containing 10 mM glucose, 20 mM ACP-Na₂, 0.5 mM ATP, 5 mM Mg²⁺, 50 mM potassium phosphate buffer (pH 7.0), 4.856 U glkase and 3.632 U ACKase were put into 37 °C water bath for 1 h.
authors.affiliation: State Key Lab of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China.
authors.forename: Bingkun
authors.initials: B
authors.lastname: Yan
authors.forename: Qingbao
authors.initials: Q
authors.lastname: Ding
authors.forename: Ling
authors.initials: L
authors.lastname: Ou
authors.forename: Zhi
authors.initials: Z
authors.lastname: Zou
mesh_terms: D000084:Acetate Kinase / Q000235:genetics / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D005941:Glucokinase / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D019298:Glucose-6-Phosphate / Q000378:metabolism*; D060847:Metabolic Engineering; D010755:Organophosphates / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D010755:Organophosphates; D011994:Recombinant Proteins; D019298:Glucose-6-Phosphate; C011632:acetyl phosphate; D000255:Adenosine Triphosphate; D005941:Glucokinase; D000084:Acetate Kinase; D005947:Glucose