Pyruvate kinase has been used in the preparation of cell free extract from BL21 CP strain of E. coli culture for highly productive cell-free protein expression. The enzyme has been used to assay the pyruvate kinase activity by incubating with eluted DAPk (Death-associated protein kinase), a serine/threonine kinase that binds and activates pyruvate kinase. It is involved in glycolysis and in gluconeogenesis. It has been used in PGM enzyme assays to determine phosphoglycerate mutase activity. It is also used to study pyruvate kinase (PK) deficiency.