反应详情

A liquid medium having the composition given in Example 1 was prepared and distributed in 50-ml portions into fifty 500-ml Sakaguchi flasks and steam-sterilized at 120° C. for 20 minutes. The contents of each flask was aseptically inoculated with 1 ml of a fluid culture obtained by cultivating Trichosporon fermentans ATCC 10675 in the same manner as in Example 1, and shake-culture was carried out at 30° C. for 24 hours. Cells were harvested from the resultant culture fluids by centrifugation and suspended in 500 ml of 100 mM phosphate buffer (pH 6.5). To this suspension, there were added 5 g of N-benzyl-3-pyrrolidinone, 10 g of glucose, 275 mg of oxidized form nicotinamide adenine dinucleotide phosphate (product of Kohjin Co.) and 1,420 units of glucose dehydrogenase (product of Amano Pharmaceutical Co.), and the reaction was allowed to proceed at 30° C. with stirring for 48 hours, while the reaction mixture was maintained at pH 6.5 with 6N aqueous sodium hydroxide solution. Thereafter, the reaction mixture was extracted with 2.5 L of ethyl acetate, and the aqueous layer was further extracted with 1 L of ethyl acetate. The organic layers were combined and dehydrated over anhydrous sodium sulfate, and the solvent was then distilled off under reduced pressure. The residue was purified by subjecting the same to silica gel column chromatography (eluent: ethyl acetate/methanol=2/1) to give 2.5 g of (S)-N-benzyl-3-pyrrolidinol. The yield was 49%, the optical purity was 96% ee, the boiling point was 132 to 137° C./3 mm Hg, and the optical rotation [α]D20 was −3.73° (CH3OH, c=5). 1H-NMR δ CDCl3): 1.63-1.76 (1H, m), 2.09-2.21 (1H, m), 2.26-2.37 (1H, m), 2.51-2.64 (2H, m), 2.75-2.85 (1H, m), 3.38 (1H, brs), 3.61 (2H, s), 4.24-4.33 (1H, m), 7.19-7.37 (5H, m).

1. customwas prepared
2. customThe contents of each flask was aseptically inoculated with 1 ml of a fluid culture
3. customobtained
4. customto proceed at 30° C.
5. extractionThereafter, the reaction mixture was extracted with 2.5 L of ethyl acetate
6. extractionthe aqueous layer was further extracted with 1 L of ethyl acetate
7. distillationthe solvent was then distilled off under reduced pressure
8. customThe residue was purified