Reaction #1530503

Reaction equation

glucose

3-oxo-3-(2-thienyl)propionic acid ethyl ester

glucose

NADH

NADPH

3-Oxo-3-(2-thienyl)propionic acid ethyl ester

Tris-HCl

→

3-hydroxy-3-(2-thienyl)propionic acid ethyl ester

Solvents

2-propanol

—

reaction solution

Conditions

Workup

1. 1
   Extractionextract
2. 2
   Extraction1% polypeptone, and 0.6% malt extract
3. 3
   OtherAfter completion of the culturing, the culture medium (1 ml) was collected
4. 4
   Otherto isolate the cells
5. 5
   Otherwas synthesized in accordance with the method
6. 6
   Otherthe reaction at 30° C. on a shaker
7. 7
   workup.ADDITIONwas added
8. 8
   Otherthe cells were removed by centrifugation

Procedure

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.

Source