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The HNL from the defatted meal of the seeds of  Pouteria sapota  ( Ps HNL) was extracted with water, and the aqueous extract was immobilized over celite or lyophilized and used as semi-purified  Ps HNL. The aqueous extract was mixed with celite, ratios 1:1, 1:2 and 1:4, and lyophilized. The immobilized  Ps HNLdir-celite catalysed the addition of HCN to  1b  in buffer saturated-DIPE (microaqueous system) with high enantioselectivity in the first cycle but diminished in the next two cycles, best results were obtained with the ratio 1:4. The reaction with  Ps HNLdir-celite in biphasic medium (5% citrates buffer) improved the conversion compared with the microaqueous system, but it was still lower to that obtained with the  Ps HNL. Other way of immobilization studied was the precipitation of the enzyme with acetone and immobilization on celite, the  Ps HNLpp-celite catalysed the enantioselective addition of HCN to 2-fluoro ( 1a ), 2-chloro ( 1b ), 2-bromo ( 1c ), 2-methyl ( 1d ) and 2-nitro ( 1e ) benzaldehydes, in the biphasic medium (1:4 ratio, 5% citrates buffer). The enantiomeric excess ( ee ) of ( R )- 2a , ( R )- 2b , ( R )- 2c  and ( R )- 2d  were &gt;98% and remained during the three cycles. The  ee  of 2-nitromandelonitrile was 82% and was raised to 95% in the third cycle. Compared with the  Ps HNL, the conversion of  1a  and  1b  diminished 6% and 40%, respectively, after third cycle; for  2c , conversion was almost 10% higher in the first two cycles and diminished 19% in the third; for  2d  and  2e  the conversion was between 18–13% and 31–18% higher, respectively.

biocatalysis and biotransformation

Preparation of optically active cyanohydrins from 2-substituted benzaldehydes using a hydroxynitrile lyase from  Pouteria sapota  seeds immobilized on celite