How Can the Solvent Affect Enzyme Enantioselectivity?

Journal of the American Chemical Society

Volumn 113, Issue 8, 1991, Pages 3166-3171

  Fitzpatrick, Paul A ^a   Klibanov, Alexander M ^a  

^a MASSACHUSETTS INSTITUTE OF TECHNOLOGY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

EID: 0000626186     PISSN: 00027863     EISSN: 15205126     Source Type: Journal    
DOI: 10.1021/ja00008a054     Document Type: Article

References (56)

1. Simon, H.; Bader, J.; Gunther, H.; Neumann, S.; Thanos, J. Angew. Chem., Int. Ed. Engl. 1985, 24, 539–553.
2. Whitesides, G. M.; Wong, C.-H. Angew. Chem., Int. Ed. Engl. 1985, 24, 617–638.
3. Jones, J. B. Tetrahedron 1986, 42, 3351–3403.
4. Yamada, H.; Shimizu, S. Angew. Chem., Int. Ed. Engl. 1988, 27, 622–624.
5. Grout, D. H. G.; Christen, M. In Modern Synthetic Methods 1989; Scheffold, R., Ed.; Springer Verlag: Berlin, 1989; pp 1–114.
6. Keinan, E.; Hafeli, F. V.; Seth, K. K.; Lamed, R. J. Am. Chem. Soc. 1986, 108, 162–169.
7. Pham, V. T.; Phillips, R. S.; Ljungdahl, L. J. Am. Chem. Soc. 1989, 111, 1935–1936.
8. Pham, V. T.; Phillips, R. S. J. Am. Chem. Soc. 1990, 112, 3629–3632.
9. Bjorkling, F.; Boutelje, J.; Gatenbeck, S.; Hult, K.; Norin, T. Tetrahedron Lett. 1985, 26, 4957–4958.
10. Guanti, G.; Banfi, L.; Narisano, E.; Riva, R.; Thea, S. Tetrahedron Lett. 1986, 27, 4639–4642.
11. Lam, L. K. P.; Hui, R. A. H. F.; Jones, J. B. J. Org. Chem. 1986, 51, 2047–2051.
12. Bjorkling, F.; Boutelje, J.; Hjalmarsson, M.; Hult, K.; Norin, T. J. Chem. Soc., Chem. Commun. 1987, 1041–1042.
13. Klibanov, A. M. Trends Biochem. Soc. 1989, 14, 141–144.
14. Sakurai, T.; Margolin, A. L.; Russell, A. J.; Klibanov, A. M. J. Am. Chem. Soc. 1988, 110, 7236–7237.
15. Kitaguchi, H.; Fitzpatrick, P. A.; Huber, J. E.; Klibanov, A. M. J. Am. Chem. Soc. 1989, 111, 3094–3095.
16. Stokes, T. M.; Oehlschlager, A. C. Tetrahedron Lett. 1987, 28, 2091–2904.
17. Kitaguchi, H.; Itoh, I.; Ono, M. Chem. Lett. 1990, 1203–1206.
18. Cambou, B.; Klibanov, A. M. J. Am. Chem. Soc. 1984, 106, 2687–2692.
19. Kirchner, G.; Scollar, P.; Klibanov, A. M. J. Am. Chem. Soc. 1985, 107, 7072–7076.
20. For reviews, see: Dordick, J. S. Enzyme Microb. Technol. 1989, 11, 194–211.
21. Chen, C.-S.; Sih, C. J. Angew. Chem., Int. Ed. Engl. 1989, 28, 695–707.
22. Klibanov, A. M. Acc. Chem. Res. 1990, 23, 114–120.
23. Zaks, A.; Klibanov, A. M. J. Biol. Chem. 1988, 263, 3194–3201.
24. Kanerva, L. T.; Klibanov, A. M. J. Am. Chem. Soc. 1989, 111, 6864–6865.
25. Adams, K. A. H.; Chung, S.-H.; Klibanov, A. M. J. Am. Chem. Soc. 1990, 112, 9418–9419.
26. Burke, P. A.; Smith, S. O.; Bachovchin, W. W.; Klibanov, A. M. J. Am. Chem. Soc. 1989, 111, 8290–8291.
27. A useful chiral reagent: Top, S.; Jaouen, G. J. Org. Chem. 1981, 46, 78–82.
28. First proposed as an acyl donor for transesterification reactions by: Degueil-Castaing, M.; DeJeso, B.; Drouillard, S.; Maillard, B. Tetrahedron Lett. 1987, 28, 953–954.
29. This solvent has been successfully used as the reaction medium for subtilisin-catalyzed reactions before.10
30. Fersht, A. Enzyme Structure and Mechanism, 2nd ed.; Freeman: New York, 1985; Chapter 8.
31. It has been previously established10 that subtilisin-catalyzed transesterifications in organic solvents follow the compulsory order scheme without ternary complexes. This mechanism involves formation of a noncovalent enzyme-ester complex, followed by its transformation to an acyl-subtilisin intermediate with the concomitant release of alcohol product. The acylsubtilisin then reacts with the exogenous nucleophile (an alcohol) to form another binary complex, which then yields the new ester and the free enzyme. In this study, subtilisin-catalyzed reaction 1 was kinetically investigated by varying the concentration of 1 in the range from 10 to 70 mM. An adherence to the aforementioned kinetic scheme was observed; e.g.,10 linear dependence of the reciprocal initial rates vs reciprocal 1 concentrations was obtained. The slopes of these dependencies afford kcat/Miaicohoi) (denoted as simply kaJKy throughout this paper) values calculated as described in the Experimental Section. Note that these kal/KM's are bimolecular rate constants for the reaction of the nucleophile (1) with butyryl-subtilisin.
32. Throughout this study, we found that the general trends and conclusions made regarding enantioselectivities were qualitatively identical whether based on the initial rates of the enzymatic transesterification or on kM/Km values for the individual enantiomers. Although the latter parameter is, generally speaking, preferable, it takes seven times as long to measure a catM compared to an initial rate. Consequently, given the large number of experiments conducted in this work, initial rates were used to calculate enantioselectivities in most instances. However, in critical experiments km/KM values were determined as well, thus leading to a seeming discrepancy in the enantioselectivity values reported (which, of course, quantitatively depend on which parameter was used to calculate them).
33. For this alcohol, there is formally a reversal of enantioselectivity simply because chlorine has a higher order of priority than carbon in the Cahn-Ingold-Prelog system (Cahn, R. S.; Ingold, C.; Prelog, V. Angew. Chem., Int. Ed. Engl. 1966, 5, 385–415). To avoid confusion, l–s nomenclature was retained for 2-chloro-l.
34. Reichardt, C. Solvents and Solvent Effects in Organic Chemistry, 2nd ed.; VCH: Weinheim, Germany, 1988.
35. Leo, A.; Hansch, C.; Elkins, D. Chem. Rev. 1971, 71, 525–616.
36. Rekker, R. F. The Hydrophobic Fragmental Constant, Elsevier: Amsterdam, 1977.
37. Laane, C.; Boeren, S.; Vos, K.; Veeger, C. Biotechnol. Bioeng. 1987, 30, 81–87.
38. It is hardly surprising that the dipole moment and dielectric constant act in a similar manner because they are intimately related via the Debye equation (Exner, O. Dipole Moments in Organic Chemistry; Georg Thieme Publishers: Stuttgart, Germany, 1975; Chapter 1).
39. Note that a satisfactory correlation was also found between subtilisin–s enantioselectivity and the Kosower Z or ET(30) solvent polarity parameter20 (this parameter has been successfully used in a recent elegant study (Smithrud, D. B.; Diederich, F. J. Am. Chem. Soc. 1990, 112, 339–343) to rationalize the strength of host-guest complexation in different organic solvents). However, this solvent polarity parameter was not used in our analysis because it usually correlates with dielectric constant and dipole moment20 but, in contrast to the latter two, is purely empirical and thus does not directly afford mechanistic interpretations.
40. Schulz, G. E.; Schirmer, R. H. Principles of Protein Structure; Springer Verlag: New York, 1979; Chapter 3.
41. Creighton, T. E. Proteins. Structures and Molecular Principles; Freeman: New York, 1983;
42. Chapter 4. Burley, S. K.; Petsko, G. A. Adv. Protein Chem. 1988, 39, 125–186.
43. Rupley, J. A.; Gratton, E.; Careri, G. Trends Biochem. Sci. 1983, 8, 18–22.
44. Finney, J. L.; Poole, P. L. Comments Mol. Cell. Biophys. 1984, 2, 129–151.
45. Zaks, A.; Klibanov, A. M. J. Biol Chem. 1988, 263, 8017–8021.
46. Kitaguchi, H.; Klibanov, A. M. J. Am. Chem. Soc. 1989, 111, 9272–9273.
47. Philipp, M.; Bender, M. L. Mol. Cell. Biochem. 1983, 51, 5–32.
48. Polgar, L.; Bender, M. L. Biochemistry 1967, 6, 610–620.
49. Greenspan, L. J. Res. Natl. Bur. Stand., Sect. A 1977, 81A, 1, 89–96.
50. Zaks, A.; Klibanov, A. M. Science 1984, 224, 1249–1251.
51. Hirata, H.; Higuchi, K.; Yamashita, T. J. Biotechnol. 1990, 14, 157–167.
52. Goldberg, M.; Thomas, D.; Legoy, M. -D. Eur. J. Biochem. 1990, 190, 603–609.
53. Sonntag, N. O. V. Chem. Rev. 1953, 52, 237–416.
54. The water content in organic solvents was measured by the optimized Fischer titration: Laitinen, A.; Harris, W. E. Chemical Analysis, 2nd ed.; Mc-Graw Hill: New York, 1975; pp 361–363.
55. Sauer, J. C. In Organic Syntheses, Wiley: New York, 1963; Collect. Vol. IV, p 561.
56. This work was financially supported by Grants GM39794 from the National Institutes of Health and CBT-8710106 from the National Science Foundation.