Kinetic resolution of chiral amines with omega-transaminase using an enzyme-membrane reactor.
A kinetic resolution process for the production of chiral amines was developed using an enzyme-membrane reactor (EMR) and a hollow-fiber membrane contactor with (S)-specific omega-transaminases (omega-TA) from Vibrio fluvialis JS17 and Bacillus thuringiensis JS64. The substrate solution containing racemic amine and pyruvate was recirculated through the EMR and inhibitory ketone product was selectively extracted by the membrane contactor until enantiomeric excess of (R)-amine exceeded 95%. Using the reactor set-up with flat membrane reactor (10-mL working volume), kinetic resolutions of alpha-methylbenzylamine (alpha-MBA) and 1-aminotetralin (200 mM, 50 mL) were carried out. During the operation, concentration of ketone product, i.e., acetophenone or alpha-tetralone, in a substrate reservoir was maintained below 0.1 mM, suggesting efficient removal of the inhibitory ketone by the membrane contactor. After 47 and 32.5 h of operation using 5 U/mL of enzyme, 98.0 and 95.5% ee of (R)-alpha-MBA and (R)-1-aminotetralin were obtained at 49.5 and 48.8% of conversion, respectively. A hollow-fiber membrane reactor (39-mL working volume) was used for a preparative-scale kinetic resolution of 1-aminotetralin (200 mM, 1 L). After 133 h of operation, enantiomeric excess reached 95.6% and 14.3 g of (R)-1-aminotetralin was recovered (97.4% of yield). Mathematical modeling of the EMR process including the membrane contactor was performed to evaluate the effect of residence time. The simulation results suggest that residence time should be short to maintain the concentration of the ketone product in EMR sufficiently low so as to decrease conversion per cycle and, in turn, reduce the inhibition of the omega-TA activity.
School of Chemical Engineering and Institute for Molecular Biology and Genetics, Seoul National University, Seoul, Korea.
J S
JS
Shin
B G
BG
Kim
A
A
Liese
C
C
Wandrey
D000588:Amines / Q000737:chemistry*; D007659:Ketones / Q000737:chemistry; D007700:Kinetics; D008956:Models, Chemical; D010627:Phenethylamines / Q000737:chemistry; D013764:Tetrahydronaphthalenes / Q000737:chemistry; D013997:Time Factors; D000637:Transaminases / Q000737:chemistry*
D000588:Amines; D007659:Ketones; D010627:Phenethylamines; D013764:Tetrahydronaphthalenes; C034592:1-aminotetralin; D000637:Transaminases; C033198:1-phenethylamine