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- Title
Hydroxynitrile Lyase of Wild Apricot ( Prunus armeniaca L.): Purification, Characterization and Application in Synthesis of Enantiopure Mandelonitrile.
- Authors
Asif, Mohammad; Bhalla, Tek
- Abstract
Hydroxynitrile lyases (HNLs) are increasingly finding application in synthesis of enantiomerically pure cyanohydrins. Cyanohydrins are important intermediates in the production of pharmaceuticals and agrochemicals. In the present studies seeds of wild apricot ( Prunus armeniaca L.) have emerged as potential source of hydroxynitrile lyase. The HNL of wild apricot (ParsHNL) was purified 8.1 fold and 18.2 % yield with a specific activity of 141 units mg protein. The SDS-PAGE of the enzyme revealed that it consists of subunits of 40 and 37 kDa. However, the molecular weight of holoenzyme was assessed to be 360 kDa. The enzyme showed maximum activity in 0.1 M sodium-citrate buffer having pH 4.75 at 25 °C. Thermostability studies revealed that this HNL showed activity up to 70 °C temperature and quite stable up to 50 °C. Activation energy of ParsHNL was calculated to be 37.83 kJ mol. This enzyme has K of 3.76 mM, V of 188.4 µmol mg min and k of 1130.4 s using mandelonitrile as substrate while for reverse reaction using benzaldehyde as substrate it showed K of 16.1 mM, V of 7.21 µmol mg min and k of 43.3 s. Synthesis of mandelonitrile was carried out using ParsHNL and finally 8.88 mmole (1.184 g) of mandelonitrile was recovered which corresponded to 89 % molar conversion with 96 % ee for R-mandelonitrile. The yield of mandelonitrile was 411 µmol mgh. These results indicated that ParsHNL has very high potential for synthesis of cyanohydrins and can be used for the production of enantiopure cyanohydrins. Graphical Abstract:
- Subjects
CYANOHYDRINS; APRICOT; NITRILE synthesis; INTERMEDIATES (Chemistry); AGRICULTURAL chemicals; TEMPERATURE effect
- Publication
Catalysis Letters, 2016, Vol 146, Issue 6, p1118
- ISSN
1011-372X
- Publication type
Article
- DOI
10.1007/s10562-016-1725-6