US 12,319,947 B2
Method for preparing (S)-nicotine by reduction
Wenqing Lin, Chongqing (CN); Hongjie Zheng, Chongqing (CN); Xiaobo Liu, Chongqing (CN); Zecong Chen, Chongqing (CN); Lingyu Li, Chongqing (CN); Qingjun Zhou, Chongqing (CN); Songhe Wang, Chongqing (CN); Yongtang Yue, Chongqing (CN); Jicheng Hu, Chongqing (CN); Yue Zhang, Chongqing (CN); and Shanshan Miao, Chongqing (CN)
Assigned to PORTON PHARMA SOLUTIONS LTD., Chongqing (CN)
Filed by Porton Pharma Solutions Ltd., Chongqing (CN)
Filed on Nov. 28, 2023, as Appl. No. 18/522,203.
Application 18/522,203 is a continuation of application No. PCT/CN2022/095423, filed on May 27, 2022.
Claims priority of application No. 202110594149.7 (CN), filed on May 29, 2021; and application No. 202210575265.9 (CN), filed on May 24, 2022.
Prior Publication US 2024/0124908 A1, Apr. 18, 2024
Int. Cl. C12P 17/12 (2006.01); C07D 401/04 (2006.01)
CPC C12P 17/12 (2013.01) [C07D 401/04 (2013.01); C12Y 101/01047 (2013.01)] 10 Claims
 
1. A method for preparing (S)-nicotine by reduction, the method comprising:
conducting a reduction process on an alkene compound shown in Formula I and/or an iminium cation compound shown in Formula II

OG Complex Work Unit Chemistry
thereby producing (S)-nicotine;
wherein
the reduction process is performed using a biocatalytic method;
the biocatalytic method comprises: in a coenzyme cycling system, catalytically reducing the alkene compound shown in Formula I and/or the iminium cation compound shown in Formula II by an imine reductase, thereby producing (S)-nicotine;
the coenzyme cycling system further comprises a coenzyme, glucose, and glucose dehydrogenase;
the imine reductase comprises an amino acid sequence selected from SEQ ID NOs: 2-4, 6, and 12-22;
the reduction process is carried out within a temperature range of 15 to 45° C.; and
the reduction process is carried out within a pH range of 6.0 to 8.0.