Table 1.
Entry | Added concentration of 1 a | Addition of | Concentration of 3 (µM) | ee value (%) |
---|---|---|---|---|
1 | 1 x 1 mMb | – | 6 ± 5 | 74 [(+)-3a] |
2 | 1 x 2.5 mMb | – | 32 ± 8 | 88 [(+)-3a] |
3 | 1 x 10 mMb | – | 995 ± 119 | 4 [(+)-3a] |
4 | 10 x 2.5 mMb | 1 every 1 h | 2730 ± 10 | 25 [(+)-3a] |
5 | 10 x 1 mMb | 1 every 1 h | 1030 ± 70 | 37 [(+)-3a] |
6 | 5 x 1 mMb | 1 every 2 h | 190 ± 20 | 44 [(+)-3a] |
7 | 3 x 1 mMb | 1 every 4 h | 63 ± 9 | ≥99 [(+)-3a] |
8 | 1 x 10 mMc,d | – | 1472 ± 16 | 1 [(+)-3a] |
9 | 1 x 10 mMc,e | C41AtPrR2 | 822 ± 44 | 97 [(+)-3a] |
10 | 1 x 10 mMc,f | C41AtPrR2 | 876 ± 21 | 98 [(+)-3a] |
11 | 1 x 10 mMc,g | C41FiPLR | 610 ± 19 | 97 [(−)-3b] |
12 | 1 x 10 mMc,e | C41FiPLR | 456 ± 19 | 95 [(−)-3b] |
13 | 1 x 10 mMc,h | C41FiPLR | 434 ± 40 | 92 [(−)-3b] |
All reaction conditions tested yielded 100 % conversion of eugenol 1
aReaction conditions: reaction buffer (50 mM KPi-buffer, pH 7.5, 100 µM IPTG), 2 % (v/v) dimethyl sulfoxide (DMSO), concentrations of eugenol 1 as indicated. Reactions were carried out for 24 h at 25 °C, 140 rpm
bRecombinant E. coli C41(DE3) harbouring PsVAO, CgL1, and AtPrR2 resuspended in 10 ml reaction buffer with an adjusted cell wet weight (cww) of 70 g l−1
cRecombinant E. coli C41(DE3) harbouring PsVAO and CgL1 resuspended in 10 ml reaction buffer with 20 g l−1 d-glucose (cww adjusted to 70 g l−1)
dWithout addition of recombinant E. coli cells harbouring AtPrR2 or FiPLR
eAddition of recombinant E. coli cells harbouring AtPrR2 or FiPLR resuspended in 10 ml reaction buffer with 20 g l−1 d-glucose (cww adjusted to 70 g l−1) after 24 h; further incubation for 4 h
fAs reaction e, but further incubation for 8 h
gAs reaction e, but further incubation for 2 h
hAs reaction e, but further incubation for 6 h