cs7b00513_si_001.pdf (2.78 MB)
Enzyme Cascades in Whole Cells for the Synthesis of Chiral Cyclic Amines
journal contribution
posted on 2017-03-14, 00:00 authored by Lorna
J. Hepworth, Scott P. France, Shahed Hussain, Peter Both, Nicholas J. Turner, Sabine L. FlitschThe
increasing diversity of reactions mediated by biocatalysts
has led to development of multistep in vitro enzyme cascades, taking
advantage of generally compatible reaction conditions. The construction
of pathways within single whole cell systems is much less explored,
yet has many advantages. Herein we report the generation of a successful
whole cell de novo enzyme cascade for the diastereoselective and/or
enantioselective conversion of simple, linear keto acids into valuable
cyclic amine products. The pathway starts with carboxylic acid reduction
that triggers a transamination, imine formation, and subsequent imine
reduction. Construction and optimization of the system was achieved
by standard genetic manipulation and the cascade required only starting
material, amine donor, and whole cell catalyst with cofactors provided
internally by glucose metabolism. A panel of synthetic keto acids
provided access to piperidines in high conversions (up to 93%) and
enantiomeric excess (up to 93%).