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T. Ohnuki et al.
LETTER
(9) (a) Fang, K.; Schlingmann, G.; Enos, A.; Carter, G. T.
obtained with the same procedure using Circinella mus-
cae (NBRC 4457) in one step.
J. Antibiotics 2001, 54, 805. (b) Argoudelis, A. D.; Coats,
J. H. J. Antibiotics 1969, 7, 341. (c) Rutkowski, M.;
Korczak, E. Experientia 1992, 48, 600. (d) Katagiri, H.;
Yamada, H.; Mitsugi, K.; Tsunoda, T. Agric. Biol. Chem.
1964, 28, 577. (e) Asano, Y.; Mihara, Y.; Yamada, H.
J. Mol. Catal. B: Enzym. 1999, 6, 271; and references cited
therein.
In conclusion, this novel direct biotransformation has
been shown to provide a useful procedure for the prepara-
tion of the phosphates of a,a-disubstituted a-amino alco-
hol derivatives using Circinella muscae, Circinella
minor, Circinella mucoroides, and Circinella umbellate.
We successfully applied this method to the preparation of
the phosphates of various kinds of a,a-disubstituted a-
amino alcohol derivatives. More detailed applications and
further work in elucidating the biological activities will be
described in future publications.
(10) Endo, A.; Yamashita, H. J. Antibiotics 1985, 38, 328.
(11) Typical Biotransformation Procedure
Circinella muscae was cultivated at 23 °C for 2 d in
Erlenmeyer flasks containing 80 mL of the medium
consisting of 0.1% malt extract, 0.3% corn steep liquor,
1.0% glucose, 0.1% polypepton, 0.1% yeast extract, and
0.1% NaH2PO4 after seed cultivation at 23 °C for 2 d. To
each of five Erlenmeyer flasks containing the fermentation
broth, 0.8 mL of 8 (40 mg, 1.1×10–4 mol) dissolved in 0.01%
aq formic acid (4 mL) was added in order to start the bio-
transformation. The reaction was carried out at 23 °C for 3 d
on a rotary shaker at 210 rpm. The fermentation broth (400
mL) was extracted with an equal volume of acetone with 40
mL of H3PO4, and the mixture was filtered. To the filtrate,
an equal volume of distilled H2O was added, and then the
mixture was adsorbed onto a column (40 mL) of DIAION
HP-20 packed with 0.1% aq H3PO4. The column was washed
with H2O (200 mL) and eluted with 30% and 50% acetone
containing 10 mM HCOONH4 (pH 8.0). Both fractions were
combined and concentrated in vacuo and lyophilized to give
crude powder (67.8 mg). The crude powder was purified by
preparative HPLC using a Develosil ODS UG-5 (150 × 20
mm i.d., Nomura Chemical Co., Ltd., Japan) with 30% aq
MeCN containing 10 mM HCOONH4 (pH 8.0) as a mobile
phase with a flow rate of 10 mL/min. The fractions were
concentrated in vacuo and lyophilized to give 10 (34.7 mg,
70% yield) as a colorless powder. IR (KBr): 3429, 2934,
2857, 2717, 2603, 1639, 1557, 1480, 1455, 1378, 1182,
1056, 1041, 946, 915 cm–1. 1H NMR (400 MHz, CD3CO2D):
d = 7.25–7.22 (m, 2 H), 7.17–7.11 (m, 3 H), 7.07 (1 H, d,
J = 4.4 Hz), 6.04 (1 H, d, J = 4.4 Hz), 4.17 (2 H, d, J = 10.3
Hz), 3.87 (s, 3 H), 2.82–2.71 (m, 4 H), 2.63 (2 H, t, J = 7.3
Hz), 2.20–2.01 (m, 2 H), 1.75–1.63 (m, 4 H), 1.46 (s, 3 H).
MS–FAB: m/z = 421 [M – H]–.
References and Notes
(1) (a) Adachi, K.; Kohara, T.; Nakao, N.; Arita, M.; Chiba, K.;
Mishina, T.; Sasaki, S.; Fujita, T. Bioorg. Med. Chem. Lett.
1995, 5, 853. (b) Brinkmann, V.; Lynch, K. R. Curr. Opin.
Immunol. 2002, 14, 569.
(2) (a) Mandala, S.; Hajdu, R.; Bergstrom, J.; Quackenbush, E.;
Xie, J.; Milligan, J.; Thornton, R.; Shei, G.-J.; Card, D.;
Keohane, C.; Rosenbach, M.; Hale, J.; Lynch, C. L.;
Rupprecht, K.; Parsons, W.; Rosen, H. Science 2002, 296,
346. (b) Brinkmann, V.; Davis, M. D.; Heise, C. E.; Albert,
R.; Cottens, S.; Hof, R.; Bruns, C.; Prieschl, E.; Baumruker,
T.; Hiestand, P.; Foster, C. A.; Zollinger, M.; Lynch, K. R.
J. Biol. Chem. 2002, 277, 21453.
(3) Hinterding, K.; Albert, R.; Cottens, S. Tetrahedron Lett.
2002, 43, 8095.
(4) Hinterding, K.; Cottens, S.; Albert, R.; Zecri, F.;
Buehlmayer, P.; Spanka, C.; Brinkmann, V.; Nussbaumer,
P.; Ettmayer, P.; Hoegenauer, K.; Gray, N.; Pan, S. Synthesis
2003, 1667.
(5) Hale, J. J.; Yan, L.; Neway, W. E.; Hajdu, R.; Bergstrom,
J. D.; Milligan, J. A.; Shei, G.-J.; Chrebet, G. L.; Thornton,
R. A.; Card, D.; Rosenbach, M.; Rosen, H.; Mandala, S.
Bioorg. Med. Chem. 2004, 12, 4803.
(6) Takeda, S.; Chino, M.; Kiuchi, M.; Adachi, K. Tetrahedron
Lett. 2005, 46, 5169.
(7) Kiuchi, M.; Adachi, K.; Tomatsu, A.; Chino, M.; Takeda, S.;
Tanaka, Y.; Maeda, Y.; Sato, N.; Mitsutomi, N.; Sugahara,
K.; Chiba, K. Bioorg. Med. Chem. 2005, 13, 425.
(8) Nakamura, T.; Tsuji, T.; Iio, Y.; Miyazaki, S.; Takemoto, T.;
Nishi, T. Tetrahedron: Asymmetry 2006, 17, 2781.
(12) The conversion ratio was determined by the peak-area ratios
of alcohol 8 and phosphate 10 using RP-HPLC. The HPLC
conditions were follows: mobile phase, 30% aq MeCN
containing 10 mM HCOONH4 (pH 4.0); column, Unison
UK-C18 (75 × 4.6 mm i.d., Imtakt Corp., Japan); flow rate,
0.8 mL/min; detection, UV 280 nm. Compounds 8 and 10
were eluted at tR = 6.7 and 3.2 min, respectively.
Synlett 2009, No. 6, 910–912 © Thieme Stuttgart · New York