S. G. Bell, W. Zhou et al.
nOe difference measurements; the diagnostic nOe being between
the H3 and H6 protons in the cis diastereomer.
J=15.9, 1.6 Hz, 1H’; H8), 4.42 (dd, J=13.7, 5.7 Hz, 1H’; H3), 3.54 (s,
1H’; OH3), 2.17 (dd, J=12.7, 5.6 Hz, 1H’; H2a), 1.97 (dd, J=6.9,
1.6 Hz, 3H’; Me10), 1.94 (dd, J=13.7, 12.7 Hz, 1H’; H2b), 1.68 (s,
3H’; Me5), 1.42 (s, 3H’; Me1a), 1.07 (s, 3H’; Me1b); 13C NMR
(126 MHz, CDCl3): d=200.83 (C4), 197.19 (C7), 161.97 (C6), 148.57
(C9), 132.71 (C8), 127.36 (C5), 69.61 (C3), 46.28 (C2), 35.89 (C1),
29.75 (Me1b), 26.36 (Me1a), 18.89 (Me10), 13.16 (Me5).
cis-3-Hydroxy-a-ionone: tR =8.54 min; m/z 209.2; 1H NMR
(500 MHz, CDCl3): d=6.53 (dd, J=15.8, 10.1 Hz, 1H’; H7), 6.09 (d,
J=15.8 Hz, 1H’; H8), 5.62 (dt, J=2.9, 1.5 Hz, 1H’; H4), 4.32–4.21 (m,
1H’; H3), 2.49 (d, J=10.1 Hz, 1H’; H6), 2.26 (s, 3H’; Me10), 1.83 (dd,
J=13.5, 5.9 Hz, 1H’; H2a), 1.62 (dd, J=2.3, 1.8 Hz, 3H’; Me5), 1.40
(dd, J=13.5, 6.3 Hz, 1H’; H2b), 1.02 (s, 3H’; Me1a), 0.88 (s, 3H’;
Me1b); 13C NMR (126 MHz, CDCl3): d=198.22 (C9), 147.28 (C7),
135.56 (C5), 133.75 (C8), 125.96 (C4), 65.60 (C3), 54.41 (C6), 43.96
(C2), 33.99 (C1), 29.44 (Me1a), 27.33 (Me10), 24.81 (Me1b), 22.80
(Me5).
Abbreviations: ArR: oxygenase-coupled NADH-dependent ferre-
doxin reductase from N. aromaticivorans, Arx: a [2Fe–2S] ferredoxin
from N. aromaticivorans, CYP: cytochrome P450, PBS: phosphate
buffered saline, rmsd: root mean square deviation.
trans-3-Hydroxy-a-ionone: tR =8.47 min; m/z 209.2, 1H NMR
(500 MHz, CDCl3): d=6.62 (dd, J=15.8, 9.5 Hz, 1H’; H7), 6.05 (d,
J=16.1 Hz, 1H’; H8), 5.57 (s, 1H’; H4), 4.26–4.20 (m, 1H’; H3), 2.25
(d, J=13.0 Hz, 1H’; H6), 2.24 (s, 3H’; Me10), 1.66 (dt, J=15.1,
7.6 Hz, 1H’; H2a), 1.60 (t, J=1.5 Hz, 3H’; Me5), 1.38 (dd, J=12.9,
9.9 Hz, 1H’; H2b), 0.95 (s, 3H’; Me1a), 0.86 (s, 3H’; Me1b); 13C NMR
(126 MHz, CDCl3): d=198.82 (C9), 148.15 (C7), 135.36 (C5), 132.71
(C8), 126.54 (C4), 66.47 (C3), 54.38 (C6), 40.61 (C2), 35.08 (C1), 29.19
(Me1b), 27.14 (Me10), 27.04 (Me1a), 22.50 (Me5).
Acknowledgements
The work was supported by a grant from the Ministry of Science
and Technology of China Project 973 (2007CB914301) to M.B.
and W.Z., and by the Higher Education Funding Council for Eng-
land (HEFCE).
Keywords: biocatalysis · CꢀH activation · crystal structure ·
cytochrome P450 · heme proteins
3-Oxo-a-ionone: tR =8.67 min; m/z 207.2, 1H NMR (500 MHz,
CDCl3): d=6.46 (dd, J=15.8, 9.6 Hz, 1H’; H7), 5.92 (d, J=15.8 Hz,
1H’; H8), 5.67 (s, 1H’; H4), 2.50 (d, J=9.6 Hz, 1H’; H6), 2.11 (d, J=
16.8 Hz, 1H’; H2a), 2.01 (s, 3H’; Me10), 1.83 (d, J=16.9 Hz, 1H’;
H2b), 1.64 (d, J=1.3 Hz, 3H’; Me5), 0.80 (s, 3H’; Me1a), 0.74 (s, 3H’;
Me1b); 13C NMR (126 MHz, CDCl3): d=197.51 (C3), 197.01 (C9),
158.77 (C5), 143.23 (C7), 133.38 (C8), 126.22 (C4), 54.78 (C6), 46.81
(C2), 36.10 (C1), 27.33 (Me1a), 26.89 (Me10), 26.77 (Me1b), 22.94
(Me5).
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4-Hydroxy-b-damascone: tR =8.27 min; m/z 209.1, 1H NMR
(500 MHz, CDCl3): d=6.76 (dq, J=15.7, 6.9 Hz, 1H’; H9), 6.14 (dq,
J=15.7, 1.5 Hz, 1H’; H8), 3.98 (t, J=5.0 Hz, 1H’; H4), 2.01–1.94 (m,
1H’; H3a), 1.92 (dd, J=6.9, 1.6 Hz, 3H’; Me10), 1.79–1.70 (m, 1H’;
H3b), 1.69–1.62 (m, 1H’; H2a), 1.63 (s, 3H’; Me5), 1.43 (ddd, J=13.1,
7.5, 3.1 Hz, 1H’; H2b), 1.02 (s, 3H’; MeMe1a), 1.02 (s, 3H’; Me1b);
13C NMR (126 MHz, CDCl3): d=201.17 (C7), 146.76 (C9), 143.61
(C6), 134.03 (C8), 131.16 (C5), 69.07 (C4), 34.74 (C2), 34.08 (C1),
28.93 (Me1b), 28.73 (C3), 27.77 (Me1a), 18.60 (Me10), 18.09 (Me5).
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3-Hydroxy-b-damascone: tR =8.39 min; m/z 209.1, 1H NMR
(500 MHz, CDCl3): d=6.72 (dq, J=15.6, 6.9 Hz, 1H’; H9), 6.15 (ddd,
J=15.7, 3.2, 1.6 Hz, 1H’; H8), 4.12–4.02 (m, 1H’; H3), 2.35 (ddd, J=
16.9, 5.9, 1.2 Hz, 1H’; H4a), 2.02 (ddd, J=17.0, 9.6, 1.0 Hz, 1H’;
H4b), 1.92 (dd, J=6.9, 1.6 Hz, 3H’; Me10), 1.74 (ddd, J=12.1, 3.6,
1.7 Hz, 1H’; H2a), 1.54 (s, 3H’; Me5), 1.50 (t, J=12.0 Hz, 1H’; H2b),
1.15 (s, 3H’; Me1a), 0.98 (s, 3H’; Me1b); 13C NMR (126 MHz, CDCl3):
d=201.81 (C7), 146.40 (C9), 140.09 (C6), 134.58 (C8), 128.21 (C5),
65.01 (C3), 47.97 (C2), 41.02 (C4), 36.48 (C1), 29.83 (Me1b), 29.20
(Me1a), 21.17 (Me5), 18.56 (Me10).
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3,4-Epoxy-b-damascone: tR =9.1 min; m/z 207.1, 1H NMR
(500 MHz, CDCl3): d=6.76 (dq, J=15.6, 6.9 Hz, 1H’; H9), 6.15 (dq,
J=15.8, 1.6 Hz, 1H’; H8), 3.95 (dd, J=8.2, 0.4 Hz, 1H’; H4), 3.88
(ddd, J=11.9, 7.9, 3.9 Hz, 1H’; H3), 1.93 (dd, J=6.9, 1.4 Hz, 3H’;
Me10), 1.72 (td, J=12.4, 3.2 Hz, 1H’; H2a), 1.68–1.58 (m, 1H’; H2b),
1.62 (d, J=0.5, 3H’; Me5), 1.18 (s, 3H’; Me1a), 0.98 (s, 3H’; Me1b);
13C NMR (126 MHz, CDCl3): d=200.53 (C7), 147.04 (C9), 142.23
(C6), 133.95 (C8), 130.16 (C5), 76.99 (C4), 71.17 (C3), 45.04 (C2),
36.28 (C1), 30.01 (Me1b), 28.85 (Me1a), 18.63 (Me10), 16.08 (Me5).
3-Hydroxy-4-oxo-b-damascone: tR =9.3 min; m/z 223.1, 1H NMR
(500 MHz, CDCl3): d=6.75 (dq, J=15.9, 6.9 Hz, 1H’; H9), 6.19 (dq,
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ChemBioChem 2011, 12, 88 – 99