Production of glucose-6-phosphate by glucokinase coupled with an ATP regeneration system.
World J Microbiol Biotechnol
; 30(3): 1123-8, 2014 Mar.
Article
en En
| MEDLINE
| ID: mdl-24165747
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 Mg²âº, 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.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Adenosina Trifosfato
/
Glucosa-6-Fosfato
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Escherichia coli
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Ingeniería Metabólica
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Glucoquinasa
Idioma:
En
Revista:
World J Microbiol Biotechnol
Año:
2014
Tipo del documento:
Article
País de afiliación:
China