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.
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Chmura et al. (2006) studied this question.