FULL PAPERS
and filtering of the sample Then the mixture was extracted
with ethyl acetate (2ꢈ500 mL), the organic layers separated
by centrifugation (90 sec, 13000 rpm), combined and dried
over Na2SO4. Then, the ee was measured on a chiral normal
phase.
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Bioamination of (Æ)-4 (Analytical Scale)
Representative example with w-TAs from the Codexisꢀ kit:
In a 2-mL Eppendorf tube, w-TA (2.0 mg) and (Æ)-4
(2.4 mg) were added in phosphate buffer 100 mM, pH 7.5
(500 mL, 1 mM PLP, 1M i-PrNH2), and DMSO (12.5 mL).
The reaction mixture was shaken at 308C and 250 rpm for
24 h. To determine the conversion, 10 mL of the mixture
were diluted with 90 mL of Mili Q water and analysed on an
achiral reverse phase column with previous centrifuging and
filtering of the sample. Then, the reaction was quenched by
addition of 50 mL of 1N aqueous NaOH and extracted twice
with AcOEt (2ꢈ400 mL). The combined organic phases
were evaporated to dryness in a vacuum concentrator and
diluted with 1 mL of ethanol. Then, the ee was measured on
a chiral normal phase.
Bioamination of (Æ)-4 (Preparative Scale)
In a 50-mL Falcon tube, (Æ)-4 (150 mg) was dissolved in
DMSO (750 mL) and phosphate buffer 100 mM, pH 7.5
(30 mL, 1 mM PLP, 1M i-PrNH2) and ATA-P1A06 (38 mg)
were added. The reaction mixture was shaken at 308C and
250 rpm and aliquots were taken periodically to measure the
conversion. After 8 h, the reaction was stopped by addition
of AcOEt (25 mL) and extracted with 3N aqueous HCl (2ꢈ
25 mL). The combined aqueous phases were basified to
pH 12 with 10N aqueous NaOH at 08C and extracted with
AcOEt (2ꢈ25 mL). The combined organic phases were
dried over Na2SO4 and evaporated to dryness in a vacuum
concentrator to yield pure (S)-3; yield: 85%; 90% ee.
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[10] This selectivity is opposite to that predicted by the Ka-
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NH2R, see: R. J. Kazlauskas, A. N. E. Weissfloch, A. T.
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b) E. Garcꢁa-Urdiales, N. Rꢁos-Lombardꢁa, J. Mangas-
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Acknowledgements
N.R.-L. acknowledges MINECO for funding under Torres-
Quevedo program (PTQ-12-05 407).
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