Enzymatic synthesis of (R)-mandelonitrile by using cross linked enzyme aggregates of Prunus cerasifera hydroxynitrile lyase

Abstract

In this study, cross-linked enzyme aggregates (CLEA) of Prunus cerasifera hydroxynitrile lyase (PcHNL-CLEA) were prepared from seeds of Prunus cerasifera and characterized for the first time. The influence of pH and temperature on the PcHNL-CLEA lyase activity was investigated. The optimal pH and temperature for lyase activity of PcHNL-CLEA were 5.5 and 25°C, respectively. We determined Km and Vmax of PcHNL-CLEA as 4.0 mM and 22 U/g PcHNL-CLEA, respectively. The thermal stability of PcHNL-CLEA at 25 and 50°C was studied. PcHNL-CLEA retained 72% of its initial activity when incubated at 25°C and 54% of initial activity when stored at 50°C at the end of 16 hours. The PcHNLCLEA was also used for the enantiopure mandelonitrile synthesis from benzaldehyde and potassium cyanide. The yield and enantiopurity of (R)-mandelonitrile were obtained as 100 and 94% respectively.