Citation: Pan Hu, Xiaofan Wu, Yi An, Xianjing Zheng, Liang Gao, Yuan Tao, Yajiao Zhang, Zedu Huang, Fener Chen. Batch and continuous-flow asymmetric synthesis of D-pantothenic acid precursor enabled by immobilized ketoreductase mutant[J]. Chinese Chemical Letters, ;2026, 37(2): 111945. doi: 10.1016/j.cclet.2025.111945 shu

Batch and continuous-flow asymmetric synthesis of D-pantothenic acid precursor enabled by immobilized ketoreductase mutant

Pan Hu ^{a, b, c, 1} ,
Xiaofan Wu ^{a, c, 1} ,
Yi An ^{a, c} ,
Xianjing Zheng ^{a, c} ,
Liang Gao ^{a, c} ,
Yuan Tao ^{a, c} ,
Yajiao Zhang ^{a, c, *,} ,
Zedu Huang ^{a, c, *,} ,
Fener Chen ^{a, b, c, d, *,}

a.
Department of Chemistry, Engineering Center of Catalysis and Synthesis for Chiral Molecules, State Key Laboratory of Green Chemical Synthesis and Conversion, Fudan University, Shanghai 200433, China
b.
Key Laboratory of Green Chemical Engineering Process of Ministry of Education, School of Chemical Engineering and Pharmacy, Wuhan Institute of Technology, Wuhan 430205, China
c.
Shanghai Engineering Research Center of Industrial Asymmetric Catalysis of Chiral Drugs, Shanghai 200433, China
d.
Institute of Flow Chemistry and Engineering, College of Chemistry and Materials, Jiangxi Normal University, Nanchang 330022, China

zhangyajiao@fudan.edu.cn

huangzedu@fudan.edu.cn

rfchen@fudan.edu.cn

Received Date: 21 July 2025
Revised Date: 27 September 2025
Accepted Date: 10 October 2025
Available Online: 11 October 2025

Figures(6)

We report an immobilized enzyme-catalyzed batch and continuous-flow synthesis of optically pure ethyl (R)-pantothenate ((R)-PaOEt), the direct precursor of D-pantothenic acid. Firstly, a ketoreductase mutant designated as M2, carrying two-point mutations of F97L and M242F relative to the wild-type SSCR, was constructed by site-directed mutagenesis, exhibited simultaneously improved activity toward ethyl 2′-ketopantothenate (K-PaOEt) and isopropanol, and could effectively catalyze the stereoselective reduction of K-PaOEt to (R)-PaOEt by using isopropanol as the sacrificial co-substrate to regenerate NADPH. After screening six commercially available carriers, an amino resin LXTE-700 was identified as the best solid support for the immobilization of M2 via the glutaraldehyde activation method. Upon optimization of the immobilization process and reaction conditions, the fabricated immobilized enzyme M2@amino resin demonstrated excellent recyclability and reusability, with the complete conversion of K-PaOEt to (R)-PaOEt being still realized after 12 cycles of reuse. Finally, M2@amino resin-catalyzed synthesis of (R)-PaOEt was successfully implemented in continuous-flow, accomplishing a 6.3 times higher space-time yield than that with the batch synthesis (529.2 versus 84 g L^-1 d^-1). Our developed flow biocatalysis system also features an outstanding operational stability, as evidenced by the 100% conversion rate achieved after 15 consecutive days of operation.
- Asymmetric synthesis,
- Continuous flow synthesis,
- Enzyme immobilization,
- Ketoreductase,
- D-Pantothenic acid
1. [1]
  S. Wu, R. Snajdrova, J.C. Moore, K. Baldenius, U.T. Bornscheuer, Angew. Chem. Int. Ed. 60 (2021) 88–119. doi: 10.1002/anie.202006648
2. [2]
  R. Buller, S. Lutz, R.J. Kazlauskas, et al., Science 382 (2023) eadh8615. doi: 10.1126/science.adh8615
3. [3]
  S. Simić, E. Zukić, L. Schmermund, et al., Chem. Rev. 122 (2022) 1052–1126. doi: 10.1021/acs.chemrev.1c00574
4. [4]
  J.M. Bolivar, J.M. Woodley, R. Fernandez-Lafuente, Chem. Soc. Rev. 51 (2022) 6251–6290. doi: 10.1039/d2cs00083k
5. [5]
  R.A. Sheldon, S. van Pelt, Chem. Soc. Rev. 42 (2013) 6223–6235. doi: 10.1039/C3CS60075K
6. [6]
  R.C. Rodrigues, Á. Berenguer-Murcia, D. Carballares, R. Morellon-Sterling, R. Fernandez-Lafuente, Biotechnol. Adv. 52 (2021) 107821. doi: 10.1016/j.biotechadv.2021.107821
7. [7]
  J. Britton, S. Majumdar, G.A. Weiss, Chem. Soc. Rev. 47 (2018) 5891–5918. doi: 10.1039/c7cs00906b
8. [8]
  Z. Tang, Y. Oku, T. Matsuda, Org. Process Res. Dev. 28 (2024) 1308–1326. doi: 10.1021/acs.oprd.3c00405
9. [9]
  M.P. Thompson, I. Peñafiel, S.C. Cosgrove, N.J. Turner, Org. Process Res. Dev. 23 (2019) 9–18. doi: 10.1021/acs.oprd.8b00305
10. [10]
  P. De Santis, L.E. Meyer, S. Kara, React. Chem. Eng. 5 (2020) 2155–2184. doi: 10.1039/d0re00335b
11. [11]
  A. Naramittanakul, S. Buttranon, A. Petchsuk, P. Chaiyen, N. Weeranoppanant, React. Chem. Eng. 6 (2021) 1771–1790. doi: 10.1039/d1re00189b
12. [12]
  A.I. Benítez-Mateos, F. Paradisi, J. Flow Chem. 14 (2024) 211–218. doi: 10.1007/s41981-023-00283-z
13. [13]
  L.E. Meyer, M. Hobisch, S. Kara, Curr. Opin. Biotechnol. 78 (2022) 102835. doi: 10.1016/j.copbio.2022.102835
14. [14]
  Y.J. Hu, J. Chen, Y.Q. Wang, et al., Chem. Eng. J. 437 (2022) 135400. doi: 10.1016/j.cej.2022.135400
15. [15]
  P. Hu, X. Wu, Z. Huang, F. Chen, Chin. J. Pharm. 56 (2025) 421–434.
16. [16]
  C. Hu, Z. Huang, M. Jiang, et al., ACS Sustainable Chem. Eng. 9 (2021) 8990–9000. doi: 10.1021/acssuschemeng.1c01419
17. [17]
  D. Andrés-Sanz, A. Maiz-Iginitz, J.M. Bolivar, et al., Green Chem. 26 (2024) 4563–4573. doi: 10.1039/d4gc00369a
18. [18]
  Q. Chen, Y. An, M. Feng, et al., Green Chem. 24 (2022) 9508–9518. doi: 10.1039/d2gc03082a
19. [19]
  G. García-Marquina, J. Langer, M. Sánchez-Costa, et al., ACS Sustainable Chem. Eng. 10 (2022) 9899–9910. doi: 10.1021/acssuschemeng.2c02279
20. [20]
  B. Shen, R. Ding, J. Dai, et al., Green Synth. Catal. 2 (2021) 367–373.
21. [21]
  R. Greifenstein, T. Ballweg, T. Hashem, et al., Angew. Chem. Int. Ed. 61 (2022) e202117144. doi: 10.1002/anie.202117144
22. [22]
  V. Williams, Y. Cui, J. Zhao, et al., Org. Process Res. Dev. 26 (2022) 1984–1995. doi: 10.1021/acs.oprd.1c00383
23. [23]
  W. Zhang, S.F. Li, J.Q. Zhu, et al., Biochem. Eng. J. 197 (2023) 108964. doi: 10.1016/j.bej.2023.108964
24. [24]
  P. Xie, J. Lan, J. Zhou, et al., Bioresour. Bioprocess. 11 (2024) 70. doi: 10.1186/s40643-024-00786-0
25. [25]
  L. Chu, X. Zhang, J. Li, et al., Chin. Chem. Lett. 35 (2024) 108896. doi: 10.1016/j.cclet.2023.108896
26. [26]
  X. Deng, M. Fan, M. Wu, et al., Chin. Chem. Lett. 35 (2024) 108684. doi: 10.1016/j.cclet.2023.108684
27. [27]
  Z. Li, H. Yang, J. Liu, Z. Huang, F. Chen, Chem. Rec. 21 (2021) 1611–1630. doi: 10.1002/tcr.202100062
28. [28]
  W. Zhang, Z.Q. Shao, Z.X. Wang, et al., Int. J. Biol. Macromol. 274 (2024) 133264. doi: 10.1016/j.ijbiomac.2024.133264
29. [29]
  L. Qiao, Z. Luo, H. Chen, et al., Chem. Commun. 59 (2023) 7518–7533. doi: 10.1039/d3cc01474f
30. [30]
  B. Su, L. Xu, X. Xu, et al., Green Synth. Catal. 1 (2020) 150–159.
31. [31]
  X. Yue, Y. Li, D. Sang, et al., Chin. Chem. Lett. 34 (2023) 108178. doi: 10.1016/j.cclet.2023.108178
32. [32]
  Z. Guo, Z. Wu, X. Wu, et al., Green Chem. 26 (2024) 7818–7824. doi: 10.1039/d4gc01478b
33. [33]
  P. Hu, X. Wu, Y. Zhang, et al., Green Chem. 26 (2024) 2124–2134. doi: 10.1039/d3gc04344d
34. [34]
  M. Eggersdorfer, D. Laudert, U. Létinois, et al., Angew. Chem. Int. Ed. 51 (2012) 12960–12990. doi: 10.1002/anie.201205886
35. [35]
  P. Cheng, J. Wang, Y. Wu, et al., Enzyme Microb. Tech. 126 (2019) 77–85. doi: 10.1016/j.enzmictec.2019.04.001
36. [36]
  H. Zhang, X. Chen, T. Lv, et al., ACS Catal. 13 (2023) 9960–9968. doi: 10.1021/acscatal.3c01569
37. [37]
  Y. Zhang, M. Liu, Z. Yang, et al., Green Chem. 25 (2023) 3223–3235. doi: 10.1039/d2gc04894a
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沈阳化工大学材料科学与工程学院沈阳 110142