Abstract
Acyl phosphate substrate availability is critical for applicable biochemical studies. Accordingly, various synthesis methods have been developed over the years typically focused on acetyl phosphate. One of these synthesis methods was proven versatile, capable of producing assorted primary and secondary acyl phosphates with high purity as measured by NMR spectroscopy, total carbon analysis and hydroxamate assays. Biochemical activity assays using synthesized acyl phosphate substrates generate results consistent with previously published kinetic constant data for Methanosarcina thermophila acetate kinase, while kinetic constant results for new substrates are reported for both M. thermophila acetate kinase and Desulfovibrio vulgaris str. Hildenborough butyrate kinase.