Abstract
AbstractThe 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 (NH4)2SO4, 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.
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