NADPH → Ethyl 3-hydroxy-3-(thiophen-2-yl)propanoate

Participants

Reaction Steps

An aqueous solution comprising a composition of 2% glucose, 1% yeast extract, 1% polypeptone, and 0.6% malt extract was used as the medium, and Filobasidium uniguttulatum IFO 0699 was inoculated into this and aerobically cultured at 28° C. for 24 hours. After completion of the culturing, the culture medium (1 ml) was collected and centrifuged to isolate the cells. 3-Oxo-3-(2-thienyl)propionic acid ethyl ester to be used as the substrate was synthesized in accordance with the method described in EP-A-751427. Said cells were suspended in 200 μl of a reaction solution comprising 100 mM glucose, 0.24% 3-oxo-3-(2-thienyl)propionic acid ethyl ester, 0.01% NADH, 0.01% NADPH and 100 mM Tris-HCl buffer (pH 7.5) and allowed to undergo the reaction at 30° C. on a shaker. After 18 hours of the commencement of the reaction, 800 μl of 2-propanol was added thereto, the cells were removed by centrifugation, and then a sample of the supernatant was analyzed by HPLC. The analysis was carried out under the following conditions using Chiralpak AD-RH (mfd. by Daicel) as the HPLC column.

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