[Energy charge changing by pathway manipulation of adenosine triphosphate metabolism in Escherichia coli].
Endocystic metabolism of energy and material is a complex matrix. We do not know the consequence of replacing the genes controlling the metabolic fluxes. Single gene (add) deletion cannot change the direction of metabolic fluxes of adenosine. Through deleting 3 genes, namely add (encoding adenosine deaminase [EC:3.5.4.4]), deoD (encoding purine-nucleoside phosphorylase [EC:2.4.2.1]) and amn (encoding AMP nucleosidase [EC:3.2.2.4]), and introducing ado1 gene(encoding adenosine kinase in S. cerevisiae[EC:2.7.1.20]), we modified the salvage pathway of adenine metabolism, and constructed a strain named J991. Extract of J991 was analyzed by HPLC. The endocystic concentration of ATP, ADP and AMP raised 2-fold as the control, and the metabolic fluxes of adenosine are also changed. It is a new way for ATP stimulation. Multi-gene manipulation is more effective than single-gene manipulation in salvage pathway of adenine.
Gu
Zhenting
Z
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China. terry.gzt@163.com
Lv
Xiaobing
X
Zhang
Huizhan
H
D000243:Adenosine Deaminase / Q000235:genetics / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism*; D004734:Energy Metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D009699:N-Glycosyl Hydrolases / Q000235:genetics / Q000378:metabolism; D011683:Purine-Nucleoside Phosphorylase / Q000235:genetics / Q000378:metabolism
D029968:Escherichia coli Proteins; D000255:Adenosine Triphosphate; D011683:Purine-Nucleoside Phosphorylase; D009699:N-Glycosyl Hydrolases; C019699:AMP nucleosidase; D000243:Adenosine Deaminase