: During the glycolytic pathway (dehydrogenation and phosphorylation), in the presence of enzyme glyceraldehyde phosphate dehydrogenase, glyceraldehyde 3-phosphate loses hydrogen to NAD to from NADH2 and accepts inorganic phosphate to form 1:3 diphosphoglycerate. Glyceraldehyde 3 - phosphate +H3PO4+NAD+1:3 phosphoglycerate +NADH+H+ One of the two phosphates of diphos-phoglycerate is linked by high energy bond. It can synthesis ATP and form 3-phosphoglycerate by using the enzyme phosphoglycerate kinase. The direct synthesis of ATP from metabolites is called substrate level phosphorylation. 1:3 diphosphoglycerat +ADP 3 - phosphoglycerate + ATP.