Development of (R)-4-hydroxymandelonitrile synthesis in an aqueous-organic biphasic stirred tank batch reactor.
A relatively new hydroxynitrile lyase-catalyzed reaction was optimized to be suitable for rapid and efficient development of a full-scale production process. The conversion of 4-hydroxybenzaldehyde into (R)-4-hydroxymandelonitrile, catalyzed by Prunus amygdalus hydroxynitrile lyase, was carried out in a biphasic system of aqueous buffer (pH 5.5) and methyl tert-butyl ether and is described with a process model. The process model consists of a description of the reaction kinetics, mass transfer kinetics, and mass balances for both the aqueous and the organic phase. Values are determined for the equilibrium constant, the enzyme kinetic parameters, the lumped mass transfer coefficient for benzaldehyde, and the partition coefficients. By using estimated prices of enzyme and reactor use, the optimum aqueous phase volume fraction and required enzyme concentration were calculated at a temperature of 20 degrees C for a batch-operated stirred tank reactor. According to the process model it was possible to convert 90% of the 4-hydroxybenzaldehyde into (R)-4-hydroxymandelonitrile with 95% enantiomeric excess. The price optimum for this reaction was found at an aqueous phase volume of 17% of the total volume. The required enzyme concentration to meet the targets was 28.6 g/L aqueous phase. At the predicted optimum, the synthesis was performed experimentally and the results were in accordance with the simulation regarding the extent of conversion and the enantiomeric excess.
Delft University of Technology, Kluyver Laboratory for Biotechnology, Julianalaan 67, NL-2628 BC Delft, The Netherlands.
W F
WF
Willeman
R
R
Neuhofer
I
I
Wirth
P
P
Pöchlauer
A J J
AJ
Straathof
J J
JJ
Heijnen
D000446:Aldehyde-Lyases / Q000737:chemistry*; D001547:Benzaldehydes / Q000737:chemistry*; D019149:Bioreactors; D002384:Catalysis; D003198:Computer Simulation; D006856:Hydrogen Cyanide / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008738:Methyl Ethers / Q000737:chemistry; D008956:Models, Chemical; D008958:Models, Molecular; D009570:Nitriles / Q000138:chemical synthesis*; D009754:Nuts / Q000201:enzymology; D027861:Prunus / Q000201:enzymology; D011786:Quality Control; D012680:Sensitivity and Specificity; D013696:Temperature
D001547:Benzaldehydes; D008738:Methyl Ethers; D009570:Nitriles; C014601:4-hydroxymandelonitrile; C043243:methyl tert-butyl ether; D006856:Hydrogen Cyanide; D000446:Aldehyde-Lyases; C079359:hydroxymandelonitrile lyase; C011483:4-hydroxybenzaldehyde