Continuous flow synthesis of chiral amines in organic solvents: immobilization of E. coli cells containing both ω-transaminase and PLP.
E. coli cells containing overexpressed (R)-selective ω-transaminase and the cofactor PLP were immobilized on methacrylate beads suitable for continuous flow applications. The use of an organic solvent suppresses leaching of PLP from the cells; no additional cofactor was required after setting up the packed-bed reactor containing the biocatalyst (ω-TA-PLP). Non-natural ketone substrates were transformed in flow with excellent enantioselectivity (>99% ee). Features of this novel system include high-throughput (30-60 min residence time), clean production (no quench, workup, or purification required), high enzyme stability (the packed-bed reactor can be continuously operated for 1-10 days), and excellent mass recovery.
†Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.
Leandro H
LH
Andrade
‡Department of Chemistry, Organic and Bioorganic Chemistry, University of Graz, Heinrichstraße 28, 8010 Graz, Austria.
Wolfgang
W
Kroutil
†Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.
Timothy F
TF
Jamison
D000588:Amines / Q000138:chemical synthesis* / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism*; D007659:Ketones; D007700:Kinetics; D015394:Molecular Structure; D011732:Pyridoxal Phosphate / Q000378:metabolism*; D012997:Solvents; D000637:Transaminases / Q000378:metabolism*
D000588:Amines; D007659:Ketones; D012997:Solvents; D011732:Pyridoxal Phosphate; D000637:Transaminases