Mutations in distant residues moderately increase the enantioselectivity of Pseudomonas fluorescens esterase towards methyl 3-bromo-2-methylpropanoate and ethyl 3-phenylbutyrate

Chemistry - A European Journal

Volumn 9, Issue 9, 2003, Pages 1933-1939

  Horsman, Geoff P ^a   Liu, Andrew M F ^a   Henke, Erik ^b   Bornscheuer, Uwe T ^b   Kazlauskas, Romas J ^a  

^a MCGILL UNIVERSITY   (Canada)

^b UNIVERSITY OF GREIFSWALD   (Germany)

Author keywords

Directed evolution; Enantioselectivity; Hydrolases; Mutagenesis; Screening

Indexed keywords

ENZYMES; ESTERS; MUTAGENESIS;

CHIRAL SYNTHONS;

MICROORGANISMS;

ASPARAGINE; BUTYRIC ACID DERIVATIVE; ESTERASE; ETHYL 3 PHENYLBUTYRATE; LEUCINE; METHYL 3 BROMO 2 METHYLPROPANOATE; PROPIONIC ACID DERIVATIVE; THREONINE; UNCLASSIFIED DRUG;

ARTICLE; BINDING SITE; CHIRALITY; ENANTIOSELECTIVITY; ENZYME ACTIVE SITE; ENZYME SUBSTRATE COMPLEX; GENE MUTATION; MOLECULAR MODEL; MUTAGENESIS; MUTANT; NONHUMAN; POINT MUTATION; PSEUDOMONAS FLUORESCENS; WILD TYPE;

BINDING SITES; DIRECTED MOLECULAR EVOLUTION; ESTERASES; FLUORESCENT DYES; MODELS, MOLECULAR; MOLECULAR CONFORMATION; MUTAGENESIS; MUTATION; PHENYLBUTYRATES; PROPIONATES; PSEUDOMONAS FLUORESCENS; STEREOISOMERISM;

EID: 0038814072     PISSN: 09476539     EISSN: None     Source Type: Journal    
DOI: 10.1002/chem.200204551     Document Type: Article

References (82)

1. a) R. Chen, Trends Biotechnol. 2001, 19, 13-14;
2. b) U. T. Bornscheuer, M. Pohl, Curr. Opin. Chem. Biol. 2001, 5, 137-143;
3. c) K.-E. Jaeger, T. Eggert, A. Eipper, M. T. Reetz, Appl. Microbiol. Biotechnol. 2001, 55, 519-530;
4. d) I. P. Petrounia, F. H. Arnold, Curr. Opin. Biotechnol. 2000, 11, 325-330;
5. e) F. H. Arnold, Chem. Eng. Sci. 1996, 51, 5091-5102;
6. f) F. H. Arnold, Acc. Chem. Res. 1998, 31, 125-131.
7. K. Liebeton, A. Zonta, K. Schimossek, M. Nardini, D. Lang, B. W. Dijkstra, M. T. Reetz, K.-E. Jaeger, Chem. Biol. 2000, 7, 709-718.
8. a) O. May, P. T. Nguyen, F. H. Arnold, Nature Biotechnol. 2000, 18, 317-3201;
9. b) E. Henke, U. T. Bornscheuer, Biol. Chem. 1999, 380, 1029-1033.
10. D. Rotticci, J. Rotticci-Mulder, S. Denman, T. Norin, K. Hult, ChemBioChem 2001, 2, 766-770.
11. A. Magnusson, K. Hult, M. Holmquist, J. Am. Chem. Soc. 2001, 123, 4354-435.
12. R. Kazlauskas, Science 2001, 293, 2277-2279.
13. a) D. Wahler, J.-L. Reymond, Curr. Opin. Chem. Biol. 2001, 5, 152-158;
14. b) M. T. Reetz, Angew. Chem. 2001, 113, 282-320; Angew. Chem. Int. Ed. 2001, 40, 284-310.
15. b) M. T. Reetz, Angew. Chem. 2001, 113, 282-320; Angew. Chem. Int. Ed. 2001, 40, 284-310.
16. a) L. E. Janes, R. J. Kazlauskas, J. Org. Chem. 1997, 62, 4560-4561;
17. b) L. E. Janes, A. C. Löwendahl, R. J. Kazlauskas, Chem. Eur. J. 1998, 4, 2317-2324;
18. c) L. E. Janes, A. Cimpoia, R. J. Kazlauskas, J. Org. Chem. 1999, 64, 9019-9029;
19. d) L. E. Janes, A. C. Löwendahl, R. J. Kazlauskas (McGill) PCT/US99/14448, 1999.
20. a) K. D. Choi, G. H. Jeohn, J. S. Rhee, O. J. Yoo, Agric. Biol. Chem. 1990, 54, 2039-2045;
21. b) I. Pelletier, J. Altenbuchner, Microbiology 1995, 141, 459-468.
22. N. Krebsfänger, F. Zocher, J. Altenbuchner, U. T. Bornscheuer, Enzyme Microb. Technol. 1998, 22, 641-646.
23. a) N. Krebsfänger, K. Schierholz, U. T. Bornscheuer, J. Biotechnol. 1998, 60, 105-112;
24. b) F. Zocher, N. Krebsfänger, O. J. Yoo, U. T. Bornscheuer, J. Mol. Catal. B Enzym. 1998, 5, 199-202;
25. c) A. M. F. Liu, N. A. Somers, R. J. Kazlauskas, T. S. Brush, F. Zocher, M. M. Enzelberger, U. T. Bornscheuer, G. P. Horsman, A. Mezzetti, C. Schmidt-Dannert, R. D. Schmid, Tetrahedron: Asymmetry 2001, 12, 545-556.
26. V. H. M. Elferink, J. G. T. Kierkels, M. Kloosterman, J. H. Roskam (Stamicarbon B.V., Neth.) EP369553, 1990 [Chem. Abstr. 1991, 114, 41029].
27. J. A. Sikorski, D. P. Getman, G. A. Decrescenzo, B. Devadas, J. N. Freskos, H.-T. Lu, J. J. McDonald (G. D. Serarle & Co.) WO 9718205, 1997 [Chem. Abstr. 1997, 127, 65754].
28. A. Fadel, A. Khesrani, Tetrahedron: Asymmetry 1998, 9, 305-320.
29. A. Tait, E. Colorni, M. Di Bella, Tetrahedron: Asymmetry 1996, 7, 2703-2706.
30. B. A. Czeskis, A. M. Moissenkov, J. Chem. Soc. Perkin Trans. 1 1989, 1353-1354.
31. T. Kogure, E. L. Eliel, J. Org. Chem. 1984, 49, 576-578.
32. a) R. Dharanipragada, E. Nicolas, G. Toth, V. J. Hruby, Tetrahedron Lett. 1989, 30, 6841-6844;
33. b) E. Nicolas, R. Dharanipragada, G. Toth, V. J. Hruby, Tetrahedron Lett. 1989, 30, 6845-6848.
34. a) A. D. Reed, M. F. Gross, S. Beaudoin, S. (Icagen, Inc., USA) WO 008191, 2002 [Chem. Abstr. 2002, 136, 151005];
35. b) J. Burrows, J. Cumming, T. McInally (AstraZeneca AB, Swed.) WO 066525, 2001 [Chem. Abstr. 2001, 135, 371644];
36. c) D. E. Clark, P. R. Eastwood, N. V. Harris, C. McCarthy, A. D. Morley, S. D. Pickett (Aventis Pharma Limited, UK) WO 049005, 2000 [Chem. Abstr. 2000, 133, 177171].
37. A.-M. Weidler, G. Bergson, Acta Chem. Scand. 1964, 18, 1483-1486.
38. G. Varadharaj, K. Hazell, C. D. Reeve, Tetrahedron: Asymmetry 1998, 9, 1191-1195.
39. a) A. Cipiciani, F. Fringuelli, O. Piermatti, F. Pizzo, R. Ruzziconi, J. Org. Chem. 2002, 67, 2665-2670;
40. b) K. Totani, S. Asano, K.-I. Takao, K.-I. Tadano, Synlett 2001, 1772-1776;
41. c) S. Sakuma, M. Sakai, R. Itooka, N. Miyaura, J. Org. Chem. 2000, 65, 5951-5955;
42. d) G. P. Lutz, H. Du, D. J. Gallagher, P. Beak, J. Org. Chem. 1996, 61, 4542-4554;
43. e) S. G. Davies, H. J. Sanganee, Tetrahedron: Asymmetry 1995, 6, 671-674;
44. f) C. Fang, T. Ogawa, H. Suemune, K. Sakai, Tetrahedron: Asymmetry 1991, 2, 389-398;
45. g) K. Soai, H. Machida, N. Yokota, J. Chem. Soc. Perkin Trans. 1 1987, 1909-1914;
46. h) S. G. Pyne, J. Org. Chem. 1986, 51, 81-87;
47. i) G. Helmchen, G. Wegner, Tetrahedron Lett. 1985, 26, 6051-6054;
48. j) W. Oppolzer, H. J. Loeher, Helv. Chim. Acta 1981, 64, 2808-2811;
49. k) T. Mukaiyama, N. Iwasawa, Chem. Lett. 1981, 913-916;
50. l) A. I. Meyers, R. K. Smith, C. E. Whitten, J. Org. Chem. 1979, 44, 2250-2256.
51. The 288 colonies represent approximately one fourth of the potential number of active mutants. Assuming that the mutator strain introduced one amino acid substitution in each gene (a good assumption based on the sequences of the isolated mutants), then the total number of possible mutants for this 272 amino acid protein is approximately 19 × 272 = 5168. However, the actual number is smaller because of the low mutation rate. Changing only one nucleotide in a codon gives on average only 5.7 possible amino acids, not 19, so the total number of likely mutants is 5.7 × 272 = 1550. We prescreened the colonies for catalytic activity on tributyrin plates and only picked active colonies. We estimate that 20% of the colonies were inactive. Thus, we estimate that our library contained ≈1200 catalytically active single amino acid mutants and the 288 colonies screened represent ≈23% of those available. This number further assumes that the colonies did not contain any wild type or duplicate mutants.
52. K. Wada, Y. Wada, F. Ishibashi, T. Gojobori, T. Ikemura, Nucleic Acids Res. 1992, 20, 2111-2118.
53. C. S. Chen, Y. Fujimoto, G. Girdaukas, C. J. Sih, J. Am. Chem. Soc. 1982, 104, 7294-7299.
54. K. Miyazaki, F. H. Arnold, J. Mol. Evol. 1999, 49, 716-720.
55. The degenerate primer is a mixture of 32 primers, each being a permutation of the degenerate codon NNK, where N denotes any of the four nucleotides and K denotes either guanosine or thymidine.
56. Nln(1 - 1/n) = ln(1 - P), where N is the number of colonies sampled, n is the number of possible codons and P is the probability. M.S. Warren, S. J. Benkovic, Protein Eng. 1997, 10, 63-68.
57. N. Guex, A. Diemand, M. C. Peitsch, Trends Biochem. Sci. 1999, 24, 364-367.
58. H. J. Hecht, H. Sobek, T. Haag, O. Pfeifer, K. H. van Pée, Nat. Struct. Biol. 1994, 1, 532-537.
59. B. Hofmann, S. Tölzer, I. Pelletier, J. Altenbuchner, K. H. van Pée, H. J. Hecht, J. Mol. Biol. 1998, 279, 889-900.
60. A. N. Shevelkova, Y. I. Sal'nikov, N. L. Kuz'mina, A. D. Ryabov, FEBS Lett. 1996, 383, 259-263.
61. I. Pelletier, J. Altenbuchner, R. Mattes, Biochim. Biophys. Acta 1995, 1250, 149-157.
62. a) C. Evrard, J. Fastrez, P. Soumillion, P. FEBS Lett. 1999, 460, 442-446;
63. b) M. Gidh-Jain, J. Takeda, L. Z. Xu, A. J. Lange, N. Vionnet, M. Stoffel, P. Froguel, G. Velho, F. Sun, D. Cohen, P. Patel, Y. D. Lo, A. T. Hattersley, H. Luthman, A. Wedell, R. S. Charles, R. W. Harrison, I. T. Weber, G. I. Bell, S. J. Pilkis, Proc. Natl. Acad. Sci. USA 1993, 90, 1932-1936;
64. c) C. A. Abbott, G. W. McCaughan, M. D. Gorrell, FEBS Lett. 1999, 458, 278-284;
65. d) A.R. Fersht, J.W. Knill-Jones, H. Bedouelle, G. Winter, Biochemistry 1988, 27, 1581-1587.
66. a) S. Hong, R. J. Spreitzer, J. Biol. Chem. 1997, 272, 11114-11117;
67. b) S. Sever, K. Rogers, M. J. Rogers, C. Carter, Jr., D. Soell, Biochemistry 1996, 35, 32-40;
68. c) J.L.E. Dean, H. Colfen, S.E. Harding, D. W. Rice, P. C. Engel, Eur. Biophys. J. 1997, 25, 417-422.
69. R J. Walters, M. A. Parniak, B. R. Akerman, C. R. Scriver, Mol. Genet. Metab. 2000, 69, 101-110.
70. a) M. Forstner, A. Müller, D. Rognan, M. Kriechbaum, T. Wallimann, Protein Eng. 1998, 11, 563-568;
71. b) S. E. Jackson, A. R. Fersht, Biochemistry 1993, 32, 13909-13916.
72. review: L. Hedstrom, Biol. Chem. 1996, 377, 465-470.
73. R. A. Bonomo, C. G. Dawes, J. R. Knox, D. M. Shlaes, Biochim. Biophys. Acta 1995, 1247, 121-125.
74. a) P. Paoli, N. Taddei, T. Fiaschi, D. Veggi, G. Camici, G. Manao, G. Raugei, F. Chiti, G. Ramponi, Arch. Biochem. Biophys. 1999, 363, 349-355;
75. b) R. Chen, A. F. Greer, A. M. Dean, Eur. J. Biochem. 1997, 250, 578-582;
76. c) A. Bashir, R. N. Perham, N. S. Scrutton, A. Berry, Biochem. J. 1995, 312, 527-533.
77. Y. Hirose, K. Kariya, Y. Nakanishi, Y. Kurono, K. Achiwa, Tetrahedron Lett. 1995, 36, 1063-1066.
78. K.-E. Jaeger, B. W. Dijkstra, M. T. Reetz, Annu. Rev. Microbiol. 1999, 53, 315-51.
79. J. N. Volff, C. Eichenseer, P. Viell, W. Piendl, J. Altenbuchner, Mol. Microbiol. 1996, 21, 1037-1047.
80. U. T. Bornscheuer, J. Altenbuchner, H. H. Meyer, Biotechnol. Bioeng. 1998, 58, 554-559.
81. U. T. Bornscheuer, M. M. Enzelberger, J. Altenbuchner, H. H. Meyer, Strategies 1998, 11, 16-17.
82. U. T. Bornscheuer, J. Altenbuchner, H. H. Meyer, Bioorg. Med. Chem. 1999, 7, 2169-2173.