title: Immobilization of Escherichia coli containing ω-transaminase activity in LentiKats®.
abstract: Whole Escherichia coli cells overexpressing ω-transaminase (ω-TA) and immobilized cells entrapped in LentiKats® were used as biocatalysts in the asymmetric synthesis of the aromatic chiral amines 1-phenylethylamine (PEA) and 3-amino-1-phenylbutane (APB). Whole cells were permeabilized with different concentrations of cetrimonium bromide (CTAB) and ethanol; the best results were obtained with CTAB 0.1% which resulted in an increase in reaction rate by 40% compared to the whole cells. The synthesis of PEA was carried out using isopropyl amine (IPA) and L-alanine (Ala) as amino donors. Using whole cell biocatalysis, the reaction with IPA was one order of magnitude faster than with Ala. No reaction was detected when permeabilized E. coli cells containing ω-TA were employed using Ala as the amino donor. Additionally, the synthesis of APB from 4-phenyl-2-butanone and IPA was studied. Whole and permeabilized cells containing ω-TA and their immobilized LentiKats® counterparts showed similar initial reactions rates and yields in the reaction systems, indicating 100% of immobilization efficiency (observed activity/activity immobilized) and absence of diffusional limitations (due to the immobilization). Immobilization of whole and permeabilized cells containing ω-TA in LentiKats® allowed improved stability as the biocatalyst was shown to be efficiently reused for five reaction cycles, retaining around 80% of original activity.
authors.affiliation: Department of Chemical Engineering, Applied Biocatalysis Unit associated to IQAC (UAB-CSIC), School of Engineering, Universitat Autònoma de Barcelona, 08193-Bellaterra (Cerdanyola del Vallès), Catalunya, Spain.
authors.forename: Max
authors.initials: M
authors.lastname: Cárdenas-Fernández
authors.forename: Watson
authors.initials: W
authors.lastname: Neto
authors.forename: Carmen
authors.initials: C
authors.lastname: López
authors.forename: Gregorio
authors.initials: G
authors.lastname: Álvaro
authors.forename: Pär
authors.initials: P
authors.lastname: Tufvesson
authors.forename: John M
authors.initials: JM
authors.lastname: Woodley
mesh_terms: D004800:Enzymes, Immobilized / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D000637:Transaminases / Q000378:metabolism*
chemical_list: D004800:Enzymes, Immobilized; D000637:Transaminases