Engergy charge changing by pathway manipulation of adenosine triphosphate metabolism in Escherichia coli

Abstract:

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.