E. Negelein
1957
Citations
0
Influential Citations
16
Citations
Journal
Methods in Enzymology
Abstract
Publisher Summary This chapter discusses the synthesis, determination, analysis, and properties of 1,3-diphosphoglyceric Acid. The principle of synthesis of 1,3-diphosphoglyceric acid(DPGA) states that glyceraldehyde 3-phosphate (GAP), phosphate , and acetaldehyde can be catalyzed into DPGA and ethanol, which is isolated as the crystalline, sparingly soluble tetrastrychnine salt. To remove the strychnine, the recrystallized material is placed with about 10 ml. of cold water into a separatory funnel and extracted several times with cold chloroform, with enough 0.2 N NaOH added each time to keep the aqueous phase weakly alkaline. The strychninefree DPGA solution is placed at 0 ° in a vacuum desiccator over silica gel to remove dissolved chloroform and to concentrate the solution somewhat. This procedure yields 10 ml. of a solution containing 2.52 X 10 -4 mole of DPGA, or 57% of the reactive portion of starting GAP. It is essential that both enzyme preparations be pure. DPGA cannot be prepared with impure protein fractions because, on the one hand, DPGA decomposes as soon as it is formed, and on the other, added GAP may be used up by conversion to dihydroxyacetone phosphate.