Rapid screening and scale-up of transaminase catalysed reactions.
A rapid, high-throughput screening methodology has been developed for the determination of transaminase activity. This pH based, colorimetric assay can also be used to scale reactions directly from 100 microL screening scale to 25 mL development scale. Additionally, three techniques have been developed to drive transamination reactions toward complete conversion. The first method uses lactate dehydrogenase to remove the inhibitory pyruvate keto acid by-product from the reaction and drive reaction equilibrium toward the desired amine. The second method is a single enzyme system, and uses a large excess of isopropylamine to drive the transamination. Method three requires only a catalytic amount of amine donor, as an amino acid dehydrogenase is employed to regenerate the amine donor in situ using ammonia. All three systems have been demonstrated for the production of optically pure methylbenzylamine from acetophenone. An enantiomeric excess of >99% was achieved for both the R- and S-methylbenzylamine products.
Truppo
Matthew D
MD
School of Chemistry, University of Manchester, Manchester Interdisciplinary Biocentre, 131 Princess Street, Manchester, UK.
Rozzell
J David
JD
Moore
Jeffrey C
JC
Turner
Nicholas J
NJ
D000098:Acetophenones / Q000737:chemistry / Q000378:metabolism; D002384:Catalysis; D007700:Kinetics; D013237:Stereoisomerism; D000637:Transaminases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism*
D000098:Acetophenones; D000637:Transaminases; C038699:acetophenone