F2,6BP is one of the most important compounds involved in allosteric regulation of glycolysis and gluconeogenesis.

F2,6BP is made by action of the enzyme phosphofructokinase-2 (PFK2).

In this reaction, fructose-6-phosphate (F6P) is converted to F2,6BP by addition of a phosphate from ATP to position 2 of the molecule (Figure 16.7).

F2,6BP is broken down to F6P by action of the enzyme fructose-2,6-bisphosphatase, which hydrolyzes the phosphate from position 2, regenerating F6P.