Stereoselective enzymatic cyclopropanation of the aliphatic alkene 2a and 1 to obtain each of four stereoisomers
of cyclopropane product 3a with diastereoselectivies
from 89:11 to <99:1 dr and enantioselectivies from 96% to >99%
ee. Reaction conditions: whole E. coli cells in M9-N
buffer, 25 mM glucose, 10 mM 2a, direct addition of 20
mM 1 under anaerobic conditions, 5% ethanol cosolvent.
The diastereoselectivity ratio (dr) is given as cis:trans, and the enantiomeric excess (ee) is given
for the major diastereomer. Catalysts used: rhodium acetate dimer
(Rh2(OAc)4) to form the racemic authentic standard,
two variants of the engineered, serine-ligated cytochrome P450BM3 (P411-UA-V87C and P411-UA-V87F), Aeropyrum pernix protoglobin W59A Y60G F145W (ApePgb AGW), and Rhodothermus marinus nitric oxide dioxygenase Q52V (RmaNOD Q52V). Protein sequences are available in the Supporting Information. Abbreviations used: RT,
room temperature; nHex, n-hexyl.