[27] AMP- and fructose 1,6-bisphosphate-activated pyruvate kinases from Escherichia coli

摘要: Publisher Summary This chapter presents an assay method, purification, and properties of AMP- fructose 1,6-bisphosphate-activated pyruvate kinases from Escherichia coli . Two noninterconvertible forms kinase are detected in coli. Both show positive cooperative effects with respect to the substrate phosphoenolpyruvate; one them is activated by 1,6-bisphosphate inhibited ATP succinyl-CoA, whereas second AMP several intermediates hexose phosphate pathway. The approaches successfully used for continuous E. include both direct coupled methods. discusses lactate dehydrogenase assay, because it most widely requires minimal amount enzyme. different physical kinetic two allow differential crude extracts containing both. purification steps involved preparation extract diethylaminoethyl (DEAE)-cellulose chromatography. Further type 1 done following heat treatment, affinity chromatography on phosphocellulose, Sephacryl S-200 chromatography, DEAE-Sephadex II involves phosphocellulose