反应详情

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.

1. extractionextract
2. extraction1% polypeptone, and 0.6% malt extract
3. customAfter completion of the culturing, the culture medium (1 ml) was collected
4. customto isolate the cells
5. customwas synthesized in accordance with the method
6. customthe reaction at 30° C. on a shaker
7. additionwas added
8. customthe cells were removed by centrifugation