Towards a Large-Scale Asymmetric Reduction Process with Isolated Enzymes: Expression of an (S)-Alcohol Dehydrogenase in E. coli and Studies on the Synthetic Potential of this Biocatalyst

Advanced Synthesis and Catalysis

Volumn 345, Issue 1-2, 2003, Pages 153-159

  Hummel, Werner ^a   Abokitse, Kofi ^a   Drauz, Karlheinz ^b   Rollmann, Claudia ^b   Gröger, Harald ^b  

^a HEINRICH HEINE UNIVERSITY   (Germany)

^b EVONIK INDUSTRIES AG   (Germany)

Author keywords

Alcohols asymmetric catalysis; Biotransformations; Enzyme catalysis; Enzymes; Reduction

Indexed keywords

ALCOHOL; ALCOHOL DEHYDROGENASE; ALIPHATIC KETONE; AROMATIC COMPOUND; ENZYME; ESTER; KETONE; NICOTINAMIDE ADENINE DINUCLEOTIDE;

ARTICLE; BACTERIAL STRAIN; BIOCATALYST; CLONING; ENANTIOSELECTIVITY; ENZYME ISOLATION; ENZYME MECHANISM; ENZYME SYNTHESIS; ESCHERICHIA COLI; NONHUMAN; PROTEIN EXPRESSION; REDUCTION; SYNTHESIS; TIME; WILD TYPE;

EID: 0242523151     PISSN: 16154150     EISSN: None     Source Type: Journal    
DOI: 10.1002/adsc.200390001     Document Type: Article

References (38)

1. Optically active (S)-alcohols are of particular interest since commercialised drugs as well as promising new drug candidates are based on their use. For representative examples, see: a) B. A. Anderson, M. M. Hansen, A. R. Harkness, C. L. Henry, J. T. Vicenzi, M. J. Zmijewski, J. Am. Chem. Soc. 1995, 117, 12538-12539; b) R. N. Patel, Adv. Appl. Microbiol. 1997, 43, 91-140; c) A. Kumar, D. H. Ner, S. Y. Dike, Tetrahedron Lett. 1991, 32, 1901-1904; d) R. A. Holdt, S. R. Rigby, (Zeneca Ltd.), US Patent 5,580,764, 1996.
2. Optically active (S)-alcohols are of particular interest since commercialised drugs as well as promising new drug candidates are based on their use. For representative examples, see: a) B. A. Anderson, M. M. Hansen, A. R. Harkness, C. L. Henry, J. T. Vicenzi, M. J. Zmijewski, J. Am. Chem. Soc. 1995, 117, 12538-12539; b) R. N. Patel, Adv. Appl. Microbiol. 1997, 43, 91-140; c) A. Kumar, D. H. Ner, S. Y. Dike, Tetrahedron Lett. 1991, 32, 1901-1904; d) R. A. Holdt, S. R. Rigby, (Zeneca Ltd.), US Patent 5,580,764, 1996.
3. Optically active (S)-alcohols are of particular interest since commercialised drugs as well as promising new drug candidates are based on their use. For representative examples, see: a) B. A. Anderson, M. M. Hansen, A. R. Harkness, C. L. Henry, J. T. Vicenzi, M. J. Zmijewski, J. Am. Chem. Soc. 1995, 117, 12538-12539; b) R. N. Patel, Adv. Appl. Microbiol. 1997, 43, 91-140; c) A. Kumar, D. H. Ner, S. Y. Dike, Tetrahedron Lett. 1991, 32, 1901-1904; d) R. A. Holdt, S. R. Rigby, (Zeneca Ltd.), US Patent 5,580,764, 1996.
4. Optically active (S)-alcohols are of particular interest since commercialised drugs as well as promising new drug candidates are based on their use. For representative examples, see: a) B. A. Anderson, M. M. Hansen, A. R. Harkness, C. L. Henry, J. T. Vicenzi, M. J. Zmijewski, J. Am. Chem. Soc. 1995, 117, 12538-12539; b) R. N. Patel, Adv. Appl. Microbiol. 1997, 43, 91-140; c) A. Kumar, D. H. Ner, S. Y. Dike, Tetrahedron Lett. 1991, 32, 1901-1904; d) R. A. Holdt, S. R. Rigby, (Zeneca Ltd.), US Patent 5,580,764, 1996.
5. For selected recent contributions in the field of asymmetric metal-catalysed hydrogenation of ketones, see: a) M. J. Burk, W. Hems, D. Herzberg, C. Malan, A. Zanotti-Gerosa, Org. Lett. 2000, 2, 4173-4176; b) Review: R. Noyori, T. Okhuma, Angew. Chem. Int. Ed. 2001, 40, 40-73; c) T. Ohkuma, H. Takeno, Y. Honda, R. Noyori, Adv. Synth. Catal. 2001, 343, 369-375; d) T. Ohkuma, M. Koizumi, M. Yoshida, R. Noyori, Org. Lett. 2000, 2, 1749-1751.
6. For selected recent contributions in the field of asymmetric metal-catalysed hydrogenation of ketones, see: a) M. J. Burk, W. Hems, D. Herzberg, C. Malan, A. Zanotti-Gerosa, Org. Lett. 2000, 2, 4173-4176; b) Review: R. Noyori, T. Okhuma, Angew. Chem. Int. Ed. 2001, 40, 40-73; c) T. Ohkuma, H. Takeno, Y. Honda, R. Noyori, Adv. Synth. Catal. 2001, 343, 369-375; d) T. Ohkuma, M. Koizumi, M. Yoshida, R. Noyori, Org. Lett. 2000, 2, 1749-1751.
7. For selected recent contributions in the field of asymmetric metal-catalysed hydrogenation of ketones, see: a) M. J. Burk, W. Hems, D. Herzberg, C. Malan, A. Zanotti-Gerosa, Org. Lett. 2000, 2, 4173-4176; b) Review: R. Noyori, T. Okhuma, Angew. Chem. Int. Ed. 2001, 40, 40-73; c) T. Ohkuma, H. Takeno, Y. Honda, R. Noyori, Adv. Synth. Catal. 2001, 343, 369-375; d) T. Ohkuma, M. Koizumi, M. Yoshida, R. Noyori, Org. Lett. 2000, 2, 1749-1751.
8. For selected recent contributions in the field of asymmetric metal-catalysed hydrogenation of ketones, see: a) M. J. Burk, W. Hems, D. Herzberg, C. Malan, A. Zanotti-Gerosa, Org. Lett. 2000, 2, 4173-4176; b) Review: R. Noyori, T. Okhuma, Angew. Chem. Int. Ed. 2001, 40, 40-73; c) T. Ohkuma, H. Takeno, Y. Honda, R. Noyori, Adv. Synth. Catal. 2001, 343, 369-375; d) T. Ohkuma, M. Koizumi, M. Yoshida, R. Noyori, Org. Lett. 2000, 2, 1749-1751.
9. For selected recent contributions in the field of asymmetric whole-cell-biocatalytic reduction of ketones, see: a) T. Matsuda, T. Harada, K. Nakamura, Chem. Commun. 2000, 1367-1368; b) Y. Yasohara, N. Kizaki, J. Hasegawa, M. Wada, M. Kataoka, S. Shimizu, Tetrahedron: Asymmetry 2001, 12, 1713-1718; c) W. Stampfer, B. Kosjek, C. Moitzi, W. Kroutil, K. Faber, Angew. Chem. 2002, 114, 1056-1059.
10. For selected recent contributions in the field of asymmetric whole-cell-biocatalytic reduction of ketones, see: a) T. Matsuda, T. Harada, K. Nakamura, Chem. Commun. 2000, 1367-1368; b) Y. Yasohara, N. Kizaki, J. Hasegawa, M. Wada, M. Kataoka, S. Shimizu, Tetrahedron: Asymmetry 2001, 12, 1713-1718; c) W. Stampfer, B. Kosjek, C. Moitzi, W. Kroutil, K. Faber, Angew. Chem. 2002, 114, 1056-1059.
11. For selected recent contributions in the field of asymmetric whole-cell-biocatalytic reduction of ketones, see: a) T. Matsuda, T. Harada, K. Nakamura, Chem. Commun. 2000, 1367-1368; b) Y. Yasohara, N. Kizaki, J. Hasegawa, M. Wada, M. Kataoka, S. Shimizu, Tetrahedron: Asymmetry 2001, 12, 1713-1718; c) W. Stampfer, B. Kosjek, C. Moitzi, W. Kroutil, K. Faber, Angew. Chem. 2002, 114, 1056-1059.
12. For recent reviews about the biocatalytic reduction, see: a) W. Hummel, Adv. Biochem. Eng./Biotechnol. 1997, 58, 146-184; b) K. Faber, Biotransformations in Organic Chemistry, 4th edn., Springer-Verlag, Berlin, 2000, chapter 2.2.3, pp. 192-194; c) K. Nakamura, T. Matsuda, in Enzyme Catalysis in Organic Synthesis, (Eds.: K. Drauz, H. Waldmann), Weinheim, 2nd edn., VCH-Wiley, 2002, chapter 15.1.
13. For recent reviews about the biocatalytic reduction, see: a) W. Hummel, Adv. Biochem. Eng./Biotechnol. 1997, 58, 146-184; b) K. Faber, Biotransformations in Organic Chemistry, 4th edn., Springer-Verlag, Berlin, 2000, chapter 2.2.3, pp. 192-194; c) K. Nakamura, T. Matsuda, in Enzyme Catalysis in Organic Synthesis, (Eds.: K. Drauz, H. Waldmann), Weinheim, 2nd edn., VCH-Wiley, 2002, chapter 15.1.
14. For recent reviews about the biocatalytic reduction, see: a) W. Hummel, Adv. Biochem. Eng./Biotechnol. 1997, 58, 146-184; b) K. Faber, Biotransformations in Organic Chemistry, 4th edn., Springer-Verlag, Berlin, 2000, chapter 2.2.3, pp. 192-194; c) K. Nakamura, T. Matsuda, in Enzyme Catalysis in Organic Synthesis, (Eds.: K. Drauz, H. Waldmann), Weinheim, 2nd edn., VCH-Wiley, 2002, chapter 15.1.
15. For selected recent examples of enzymatic reductions under in situ cofactor regeneration which were carried out successfully on a gram scale, see: a) M. Wolberg, W. Hummel, C. Wandrey, M. Müller, Angew. Chem. 2000, 112, 4476-4478; Angew. Chem. Int. Ed. 2000, 39, 4306-4308; b) M. Wolberg, A. G. Ji, W Hummel, M. Müller, Synthesis 2001, 937-942; c) M. Wolberg, W. Hummel, M. Müller, Chem. Eur. J. 2001, 7, 4652-4571; 5d) T. Schubert, W. Hummel, M. Müller, Angew. Chem. 2002, 114, 656-659; Angew. Chem. Int. Ed. 2002, 41, 634-637.
16. For selected recent examples of enzymatic reductions under in situ cofactor regeneration which were carried out successfully on a gram scale, see: a) M. Wolberg, W. Hummel, C. Wandrey, M. Müller, Angew. Chem. 2000, 112, 4476-4478; Angew. Chem. Int. Ed. 2000, 39, 4306-4308; b) M. Wolberg, A. G. Ji, W Hummel, M. Müller, Synthesis 2001, 937-942; c) M. Wolberg, W. Hummel, M. Müller, Chem. Eur. J. 2001, 7, 4652-4571; 5d) T. Schubert, W. Hummel, M. Müller, Angew. Chem. 2002, 114, 656-659; Angew. Chem. Int. Ed. 2002, 41, 634-637.
17. For selected recent examples of enzymatic reductions under in situ cofactor regeneration which were carried out successfully on a gram scale, see: a) M. Wolberg, W. Hummel, C. Wandrey, M. Müller, Angew. Chem. 2000, 112, 4476-4478; Angew. Chem. Int. Ed. 2000, 39, 4306-4308; b) M. Wolberg, A. G. Ji, W Hummel, M. Müller, Synthesis 2001, 937-942; c) M. Wolberg, W. Hummel, M. Müller, Chem. Eur. J. 2001, 7, 4652-4571; 5d) T. Schubert, W. Hummel, M. Müller, Angew. Chem. 2002, 114, 656-659; Angew. Chem. Int. Ed. 2002, 41, 634-637.
18. For selected recent examples of enzymatic reductions under in situ cofactor regeneration which were carried out successfully on a gram scale, see: a) M. Wolberg, W. Hummel, C. Wandrey, M. Müller, Angew. Chem. 2000, 112, 4476-4478; Angew. Chem. Int. Ed. 2000, 39, 4306-4308; b) M. Wolberg, A. G. Ji, W Hummel, M. Müller, Synthesis 2001, 937-942; c) M. Wolberg, W. Hummel, M. Müller, Chem. Eur. J. 2001, 7, 4652-4571; 5d) T. Schubert, W. Hummel, M. Müller, Angew. Chem. 2002, 114, 656-659; Angew. Chem. Int. Ed. 2002, 41, 634-637.
19. For selected recent examples of enzymatic reductions under in situ cofactor regeneration which were carried out successfully on a gram scale, see: a) M. Wolberg, W. Hummel, C. Wandrey, M. Müller, Angew. Chem. 2000, 112, 4476-4478; Angew. Chem. Int. Ed. 2000, 39, 4306-4308; b) M. Wolberg, A. G. Ji, W Hummel, M. Müller, Synthesis 2001, 937-942; c) M. Wolberg, W. Hummel, M. Müller, Chem. Eur. J. 2001, 7, 4652-4571; 5d) T. Schubert, W. Hummel, M. Müller, Angew. Chem. 2002, 114, 656-659; Angew. Chem. Int. Ed. 2002, 41, 634-637.
20. For selected recent examples of enzymatic reductions under in situ cofactor regeneration which were carried out successfully on a gram scale, see: a) M. Wolberg, W. Hummel, C. Wandrey, M. Müller, Angew. Chem. 2000, 112, 4476-4478; Angew. Chem. Int. Ed. 2000, 39, 4306-4308; b) M. Wolberg, A. G. Ji, W Hummel, M. Müller, Synthesis 2001, 937-942; c) M. Wolberg, W. Hummel, M. Müller, Chem. Eur. J. 2001, 7, 4652-4571; 5d) T. Schubert, W. Hummel, M. Müller, Angew. Chem. 2002, 114, 656-659; Angew. Chem. Int. Ed. 2002, 41, 634-637.
21. [6b] it is mentioned that "many of the difficulties associated with the application of whole cells as catalyst can be avoided using isolated enzymes." As main disadvantages of (natural) whole-cell catalysts the presence of more than one alcohol dehydrogenases (leading to side reactions, and reducing the enantioselectivity), and diffusion limitations are known. Furthermore, the properties of the cofactor-regenerating enzyme FDH have been improved remarkably by protein engineering in the Kula group. Thus, a stable and efficient formate dehydrogenase is available for NAD-regeneration, see: H. Slusarczyk, S. Felber, M.-R. Kula, M. Pohl, Eur. J. Biochem. 2000, 267, 1280-1289.
22. [6b] it is mentioned that "many of the difficulties associated with the application of whole cells as catalyst can be avoided using isolated enzymes." As main disadvantages of (natural) whole-cell catalysts the presence of more than one alcohol dehydrogenases (leading to side reactions, and reducing the enantioselectivity), and diffusion limitations are known. Furthermore, the properties of the cofactor-regenerating enzyme FDH have been improved remarkably by protein engineering in the Kula group. Thus, a stable and efficient formate dehydrogenase is available for NAD-regeneration, see: H. Slusarczyk, S. Felber, M.-R. Kula, M. Pohl, Eur. J. Biochem. 2000, 267, 1280-1289.
23. [6b] it is mentioned that "many of the difficulties associated with the application of whole cells as catalyst can be avoided using isolated enzymes." As main disadvantages of (natural) whole-cell catalysts the presence of more than one alcohol dehydrogenases (leading to side reactions, and reducing the enantioselectivity), and diffusion limitations are known. Furthermore, the properties of the cofactor-regenerating enzyme FDH have been improved remarkably by protein engineering in the Kula group. Thus, a stable and efficient formate dehydrogenase is available for NAD-regeneration, see: H. Slusarczyk, S. Felber, M.-R. Kula, M. Pohl, Eur. J. Biochem. 2000, 267, 1280-1289.
24. [6b] it is mentioned that "many of the difficulties associated with the application of whole cells as catalyst can be avoided using isolated enzymes." As main disadvantages of (natural) whole-cell catalysts the presence of more than one alcohol dehydrogenases (leading to side reactions, and reducing the enantioselectivity), and diffusion limitations are known. Furthermore, the properties of the cofactor-regenerating enzyme FDH have been improved remarkably by protein engineering in the Kula group. Thus, a stable and efficient formate dehydrogenase is available for NAD-regeneration, see: H. Slusarczyk, S. Felber, M.-R. Kula, M. Pohl, Eur. J. Biochem. 2000, 267, 1280-1289.
25. a) W. Hummel, H. Schütte, E. Schmidt, C. Wandrey, M.-R. Kula, Appl. Microbiol. Biotechnol. 1987, 26, 409-416;
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28. Other prerequisites which have to be fulfilled in order to realise a valuable and large-scale feasible reduction with isolated enzymes are, e. g., high yields, high enantioselectivities, and in particular good space-time yield (so far often enzymatic reductions are typically carried out at low substrate concentrations of ca. 10 mM).
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32. [6b] NAD is about seven times cheaper compared with NADP.
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35. [13] A detailed description of these molecular biological studies covering the plasmid concept, and a new vector system in detail will be published independently from this contribution in due course; a brief experimental summary is already included in the experimental section.
36. For example, compared with the wild-type ADH, the new alcohol dehydrogenase expressed in E. coli shows a preference for the substituted acetophenones Id towards the acetoacid esters 1j. The new alcohol dehydrogenase expressed in E. coli also led to a higher activity for p-chloroacetophenone compared with p-methylacetophenone. In case of the wild-type ADH the opposite effect was observed with higher activities for the latter substrate.
37. W. Hummel, K. Abokitse, unpublished results.
38. These results will be published in detail in due course elsewhere.