Advanced Synthesis & Catalysis · 2006 · 58 citations · 0 references
Life SciencesBiochemistryChemistryBiocatalysisBioorganic ChemistryEnzymologyBiocatalyst DiscoveryBiocatalyst ScreeningCatalytic MechanismsEnzymatic CatalysisEnzyme DiscoveryEnzyme KineticsAldehydesAssayAssay OptimizationBiocatalyst Activity AssaysBiocatalyst CharacterizationCatalytic Activity AssaysHydrolytic Enzyme ActivityLyase Catalysis
Abstract The precipitation and cross‐linking into CLEAs of the hydroxynitrile lyase (E.C. 4.1.2.10) from Manihot esculenta was investigated and an optimized procedure, which involved precipitation with (NH 4 ) 2 SO 4 , was developed. It was found that a fast (photometric) assay severely underestimated the activity recovery in the CLEA due to rate‐limiting diffusion, whereas in a synthetic assay the activity recovery was up to 93 % of the starting activity. The CLEA was applied in the hydrocyanation of various aldehydes and ketones in microaqueous medium, conditions that rendered the non‐enzymatic background reaction insignificant, even with unreactive substrates.