Table 1: Synthesis of cyanohydrins with DtHNL1.

From: Enzyme discovery beyond homology: a unique hydroxynitrile lyase in the Bet v1 superfamily

EntrySubstrate 0.5 hours2 hours6 hours24 hours
conveeconveeconveeconvee
Product%%%%%%%%
1benzaldehyde → (2R)-2-hydroxy-2-phenylacetonitrileDtHNL195.199.097.494.597.985.999.561.4
Control−4.911.70.818.50.339.70.5
22-chlorobenzaldehyde → (2R)-2-(2-chlorophenyl)-2-hydroxyacetonitrileDtHNL138.689.475.692.196.688.499.671.3
Control11.3−14.019.6−5.728.7−2.359.8−1.5
33-phenylpropanal → (2R)-2-hydroxy-4-phenylbutanenitrileDtHNL150.945.884.848.394.548.193.248.0
Control18.00.436.30.253.00.080.20.2
43-phenylprop-2-enal → (2R)-2-hydroxy-4-phenylbut-3-enenitrileDtHNL15.382.346.695.288.294.598.292.6
Control−17.0−11.6−12.6−10.5
5furan-2-carbaldehyde → (2S)-furan-2-yl-hydroxyacetonitrileDtHNL195.399.398.098.998.097.698.192.9
Control10.50.020.50.435.90.171.1−0.1
61-phenylethanone → 2-hydroxy-2-phenylpropanenitrileDtHNL19.8n.d.17.3n.d.23.5n.d.22.3n.d.
Control6.66.66.97.0
  1. The synthesis of cyanohydrins was performed with 3 mg of DtHNL1. 0.5 M aldehyde or 0.3 M of ketone were mixed with 2 M HCN in TBME (final volume 1 mL), 2% v/v triisopropylbenzene was added as internal standard. To monitor the non-enzymatic formation of cyanohydrins, independent control reactions were set up at the same conditions, but omitting the enzyme (Control). Reaction conditions: pH 4.0, 10 °C and 1,000 rpm. Samples were analyzed by GC after acetylation of the product. Conversion (conv) is based on the substrate consumption. Entry 5: change of product configuration as a consequence of the Cahn–Ingold–Prelog rule. Dashes indicate that no product peak was detected. n.d., not determined.