反応 #2006457

反応方程式

sodium hydroxide

N-benzyl-3-pyrrolidinone

D-glucose

NAD+

D-alanine

pyridoxal phosphate

→

N-benzyl-3-aminopyrrolidine

反応条件

後処理

1. 1
   その他(pNTASPAG), obtained in Example 11, which
2. 2
   workup.ADDITIONwere added
3. 3
   その他During the reaction
4. 4
   その他the reaction liquid
5. 5
   workup.ALIQUOTwas sampled
6. 6
   抽出From the sample, N-benzyl-3-aminopyrrolidine was extracted by the addition of ethyl acetate

実験手順

To a flask containing 0.6 g of a substrate N-benzyl-3-pyrrolidinone, 1.85 g of D-glucose, 2.3 mg of NAD+, 1.83 g of D-alanine, and 4.0 mg of pyridoxal phosphate, 30 ml of a culture fluid of the E. coli HB101 (pNTASPAG), obtained in Example 11, which expresses TAS, PALDH, and GDH were added. The resultant product was stirred at 30° C. while being adjusted to a pH of 6.8 by drippage of 5 N aqueous solution of sodium hydroxide. During the reaction, the reaction liquid was sampled. From the sample, N-benzyl-3-aminopyrrolidine was extracted by the addition of ethyl acetate after basifying the sample with 6 N aqueous solution of sodium hydroxide. The amount of N-benzyl-3-aminopyrrolidine produced was measured by analyzing N-benzyl-3-aminopyrrolidine under the following HPLC conditions. Further, by the law of the art, N-benzyl-3-aminopyrrolidine thus obtained was acted on by 3,5-dinitrobenzyl chloride to form a dinitrobenzyl derivative. After that, the dinitrobenzyl derivative was analyzed under the following HPLC conditions, whereby the optical purity thereof was measured.

出典