Engineering Enantioselectivity in Enzyme-Catalyzed Reactions

Protein Engineering Handbook, Volume 1 & Volume 2

Volumn 1, Issue , 2011, Pages 15-46

  Kazlauskas, Romas ^a  

^a UNIVERSITY OF MINNESOTA   (United States)

Author keywords

Computer modeling; Enantioselectivity; Kinetic resolution; Lipases; Random mutagenesis; Reversing enantioselectivity; Site directed mutagenesis; X ray crystal structure

Indexed keywords

EID: 84884603107     PISSN: None     EISSN: None     Source Type: Book    
DOI: 10.1002/9783527634026.ch2     Document Type: Chapter

References (98)

1. Taylor, S.J.C., Holt, K.E., Brown, R.C., Keene, P.A. and Taylor, I.N. (2000) Choice of biocatalyst in the development of industrial biotransformations, in Stereoselective Biocatalysis (ed. R.M. Pateland), Marcel Dekker, New York, pp. 397-413 .
2. Sakowicz, R., Gold, M. and Jones, J.B. (1995) Partial reversal of the substrate stereospecifi city of an L-lactate dehydrogenase by site-directed mutagenesis . Journal of The American Chemical Society,117,2387-94 .
3. Chen, C.S., Fujimoto, Y., Girdaukas, G., and Sih, C.J. (1982) Quantitative analyses of biochemical kinetic resolutions of enantiomers . Journal of the American Chemical Society,104,7294-9 .
4. Patel, R.N., Robison, R.S., and Szarka, L.J. (1990) Stereoselective enzymic hydrolysis of 2-cyclohexyl-and 2-phenyl-1,3-propanediol diacetate in biphasic systems . Applied Microbiology and Biotechnology,34,10-14 .
5. Patel, R.N. (2000) Stereoselective biocatalysis for synthesis of some chiral pharmaceutical intermediates, in Stereoselective Biocatalysis (ed. R.M. Patel), Marcel Dekker, New York, pp. 87-130 .
6. Truppo, M.D., Pollard, D., and Devine, P. (2007) Enzyme-catalyzed enantioselective diaryl ketone reductions . Organic Letters,9,335-8 .
7. Gruber, C.C., Lavandera, I., Faber, F., and Kroutil, W. (2006) From a racemate to a single enantiomer: deracemization by stereoinversion . Advanced Synthesis Catalysis,348,1789-805 .
8. Schütte, H., Hummel, W., and Kula, M.-R. (1984) L-2-Hydroxyisocaproate dehydrogenase-a new enzyme from Lactobacillus confusus for the stereospecifi c reduction of 2-ketocarboxylic acids . Applied Microbiology and Biotechnology,19,167-76 .
9. Niefind, K., Hecht, H.J., and Schomburg, D. (1995) Crystal structure of L-2-hydroxyisocaproate dehydrogenase from Lactobacillus confusus at 2.2 A resolution. An example of strong asymmetry between subunits . Journal of Molecular Biology,251,256-81 .
10. Dengler, U., Niefind, K., Kiess, M., and Schomburg, D. (1997) Crystal structure of a ternary complex of D-2-hydroxyisocaproate dehydrogenase from Lactobacillus casei, NAD + and 2-oxoisocaproate at 1.9 Å resolution . Journal of Molecular Biology,267,640-60 .
11. Mugford, P.F., Wagner, U., Jiang, Y., Kurt Faber, K., and Kazlauskas, R.J. (2008) Enantiocomplementary enzymes: classifi cation, molecular basis for their reversed enantiopreference and prospects for mirror-image biotransformations . Angewandte Chemie-International Edition,47 .
12. Mezzetti, A., Schrag, J.D., Cheong, C.S., and Kazlauskas, R.J. (2005) Mirror-image packing in enantiomer discrimination: molecular basis for the enantioselectivity of B. cepacia lipase toward 2-methyl-3-phenyl-1-propanol . Chemistry and Biology,12,427-37 .
13. Bentley, R. (2003) Diastereoisomerism, contact points, and chiral selectivity: a four-site saga . Archives of Biochemistry and Biophysics,414,1-12 .
14. Cygler, M., Grochulski, P., Kazlauskas, R.J., Schrag, J.D., Bouthillier, F., Rubin, B., Serreqi, A.N., and Gupta, A.K. (1994) Molecular basis for the chiral preference of lipases . Journal of the American Chemical Society,116,3180-6 .
15. Vorlova, S., Bornscheuer, U.T., Gatfield, I., Hilmer, J.M., Bertram, H.J., and Schmid, R.D. (2002) Enantioselective hydrolysis of D, L-menthyl benzoate to L-(-)-menthol by recombinant Candida rugosa lipase LIP1 . Advanced Synthesis Catalysis,344,1152-5 .
16. Nishizawa, K., Ohgami, Y., Matsuo, N., Kisida, H., and Hirohara, H. (1997) Studies on hydrolysis of chiral, achiral and racemic alcohol esters with Pseudomonas cepacia lipase: mechanism of stereospecifi city of the enzyme . Journal of the Chemical Society, Perkin Transactions,2,1293-8 .
17. Tuomi, W.V., and Kazlauskas, R.J. (1999) Molecular basis for enantioselectivity of lipase from Pseudomonas cepacia toward primary alcohols. Modeling, kinetics, and chemical modification of Tyr29 to increase or decrease enantioselectivity . Journal of Organic Chemistry,64,2638-47 .
18. Kwon, D.Y., Hong, Y.-J., and Yoon, S.H. (2000) Enantiomeric synthesis of (S)-2-methylbutanoic acid methyl ester, apple fl avor, using lipases in organic solvent . Journal of Agricultural and Food Chemistry,48,524-30 .
19. Ueji, S.-I., Watanabe, K., Koshiba, T., Nakamura, M., Oh-Ishi, K., Yasufuku, Y., and Miyazawa, T. (1999) Lipase-catalyzed esterifi cation of 2-(4-substituted phenoxy)propionic acids in organic solvents: substituent effect controlling enantioselectivity toward racemic acids . Biotechnology Letters,21,865-8 .
20. Weissfloch, A.N.E., and Kazlauskas, R.J. (2001) Choosing hydrolases for enantioselective reactions involving alcohols using empirical rules, in Methods in Biotechnology, Vol. 15: Enzymes in Nonaqueous Solvents: Methods and Protocols (eds E.N. Vulfson, P.J. Halling and H.L. Holland), Humana Press, Totowa, NJ, pp. 243-59 .
21. Kazlauskas, R.J., Weissfloch, A.N.E., Rappaport, A.T., and Cuccia, L.A. (1991) A rule to predict which enantiomer of a secondary alcohol reacts faster in reactions catalyzed by cholesterol esterase, lipase from Pseudomonas cepacia, and lipase from Candida rugosa . Journal of Organic Chemistry,56,2656-65 .
22. Burgess, K., and Jennings, L.D. (1991) Enantioselective esterifi cations of unsaturated alcohols mediated by a lipase prepared from Pseudomonas . sp . Journal of the American Chemical Society,113,6129-39 .
23. Naemura, K., Ida, H., and Fukuda, R. (1993) Lipase YS-catalyzed enantioselective transesterifi cation of alcohols of bicarbocyclic compounds . Bulletin of the Chemical Society of Japan,66,573-7 .
24. Kim, M.J., and Cho, H. (1992) Pseudomonas lipases as catalysts in organic synthesis: specifi city of lipoprotein lipase . Journal of the Chemical Society, Chemical Communications,1411-13 .
25. Roberts, S.M. (1989) Use of enzymes as catalysts to promote key transformations in organic synthesis . Philosophical Transactions of the Royal Society of London. Series B,324,577-87 .
26. Janssen, A.J.M., Klunder, A.J.H., and Zwanenburg, B. (1991) Resolution of secondary alcohols by enzyme-catalyzed transesterifi cation in alkyl carboxylates as the solvent . Tetrahedron,47,7645-62 .
27. Kazlauskas, R.J., Weissfloch, A.N.E. (1997) A structure-based rationalization of the enantiopreference of subtilisin toward secondary alcohols and isosteric primary amines (Review) . Journal of Molecular Catalysis B Enzymatic,3,65-72 .
28. Zhang, M., and Kazlauskas, R. (1999) First preparation of enantiopure indane monomer, (S)-(-)-and (R)-(+)-2,3-dihydro-3-(4 '-hydroxyphenyl)-1,1,3-trimethyl-1 H-inden-5-ol, via a unique enantio-and regioselective enzymatic kinetic resolution . Journal of Organic Chemistry,64,7498-503 .
29. Sundaramoorthy, M., Terner, J., and Poulos, T.L. (1998) Stereochemistry of the chloroperoxidase active site: crystallographic and molecular-modeling studies . Chemistry and Biology,5,461-73 .
30. Holmquist, M., Hæffner, F., Norin, T., and Hult, K. (1996) A structural basis for enantioselective inhibition of Candida rugosa lipase by long-chain aliphatic alcohols . Protein Science,5,83-8 .
31. Botta, M., Cernia, E., Corelli, F., Manetti, F., and Soro, S. (1997) Probing the substrate specifi city for lipases. II. Kinetic and modeling studies on the molecular recognition of 2-arylpropionic esters by Candida rugosa and Rhizomucor miehei lipases . Biochimica et Biophysica Acta,1337,302-10 .
32. Berglund, P., Holmquist, M., and Hult, K. (1998) Reversed enantiopreference ofCandida rugosa lipase supports different modes of binding enantiomers of a chiral acyl donor . Journal of Molecular Catalysis B Enzymatic,5,283-7 .
33. Bocola, M., Otte, N., Jaeger, K.-E., Reetz, M.T., and Thiel, W. (2004) Learning from directed evolution: theoretical investigations into cooperative mutations in lipase enantioselectivity . Chembiochem: A European Journal of Chemical Biology,5,214-23 .
34. Reetz, M.T., Puls, M., Carballeira, J.D., Vogel, A., Jaeger, K.-E., Eggert, T., Thiel, W., Bocola, M., and Otte, N. (2007) Learning from directed evolution: further lessons from theoretical investigations into cooperative mutations in lipase enantioselectivity . Chembiochem: A European Journal of Chemical Biology,8,106-12 .
35. Schulz, T., Pleiss, J., and Schmid, R.D. (2000) Enantioselectivity of Pseudomonas cepacia lipase toward secondary alcohols: a quantitative model . Protein Science,9,1053-62 .
36. Phillips, R.S. (1992) Temperature effects on stereochemistry of enzymatic reactions . Enzyme and Microbial Technology,14,417-19 .
37. Overbeeke, P.L.A., Orrenius, S.C., Jongejan, J.A., and Duine, J.A. (1998) Enthalpic and entropic contributions to lipase enantioselectivity . Chemistry and Physics of Lipids,93,81-93 .
38. Overbeeke, P.L.A., Ottosson, J., Hult, K., Jongejan, J.A., and Duine, J.A. (1999) The temperature dependence of enzymatic kinetic resolutions reveals the relative contribution of enthalpy and entropy to enzymatic enantioselectivity . Biocatalysis and Biotransformation,17,61-79 .
39. Ottosson, J., Rotticci-Mulder, J.C., Rotticci, D., and Hult, K. (2001) Rational design of enantioselective enzymes requires considerations of entropy . Protein Science,10,1769-74 .
40. Ottosson, J., Fransson, L., and Hult, K. (2002) Substrate entropy in enzyme enantioselectivity: an experimental and molecular modeling study of a lipase . Protein Science,11,1462-71 .
41. Helms, V., and Wade, R.C. (1998) Computational alchemy to calculate absolute protein-ligand binding free energy . Journal of the American Chemical Society,120,2710-13 .
42. Massova, I., and Kollman, P.A. (1999) Computational alanine scanning to probe protein-protein interactions: a novel approach to evaluate binding free energies . Journal of the American Chemical Society,121,8133-43 .
43. Stanton, R.V., Perakyla, M., Bakowies, D., and Kollman, P.A. (1998) Combined ab initio and free energy calculations to study reactions in enzymes and solution-amide hydrolysis in trypsin and aqueous solution . Journal of the American Chemical Society,120,3448-57 .
44. Ke, T., and Klibanov, A.M. (1998) Insights into the solvent dependence of chymotryptic prochiral selectivity . Journal of the American Chemical Society,120,4259-63 .
45. Colombo, G., Ottolina, G., Carrea, G., Bernardi, A., and Scolastico, C. (1998) Application of structure-based thermodynamic calculations to the rationalization of the enantioselectivity of subtilisin in organic solvents . Tetrahedron: Asymmetry,9,1205-14 .
46. Savile, C.K., and Kazlauskas, R.J. (2005) How substrate solvation contributes to the enantioselectivity of subtilisin toward secondary alcohols . Journal of the American Chemical Society,127,12228-9 .
47. Leonard, V., Fransson, L., Lamare, S., Hult, K., and Graber, M. (2007) A water molecule in the stereospecifi city pocket of Candida antarctica lipase B enhances enantioselectivity towards pentan-2-ol . Chembiochem: A European Journal of Chemical Biology,8,662-7 .
48. Ottosson, J., Fransson, L., King, J.W., and Hult, K. (2002) Size as a parameter for solvent effects on Candida antarctica lipase B enantioselectivity . Biochimica et Biophysica Acta,1594,325-34 .
49. Tantillo, D.J., and Houk, K.N. (1999) Fidelity in hapten design: how analogous are phosphonate haptens to the transition states for alkaline hydrolysis of aryl esters . Journal of Organic Chemistry,64,3066-76 .
50. Colombo, G., Toba, S., and Merz, K.M. Jr (1999) Rationalization of the enantioselectivity of subtilisin in DMF . Journal of the American Chemical Society,121,3486-93 .
51. Truhlar, D.G., Gao, J., Alhambra, C., Garcia-Viloca, M., Corchado, J., Sánchez, M.L., and Villà, J. (2002) The incorporation of quantum effects in enzyme kinetics modeling . Accounts of Chemical Research,35,341-9 .
52. Pu, J., Gao, J., and Truhlar, D.G. (2006) Multidimensional tunneling, recrossing, and the transmission coeffi cient for enzymatic reactions . Chemical Reviews,106,3140-69 .
53. Zuegg, J., Hönig, H., Schrag, J.D., and Cygler, M. (1997) Selectivity of lipases: conformational analysis of suggested intermediates in ester hydrolysis of chiral primary and secondary alcohols . Journal of Molecular Catalysis B Enzymatic,3,83-98 .
54. Orrenius, C., Hæffner, F., Rotticci, D., Ohrner, N., Norin, T., and Hult, K. (1998) Chiral recognition of alcohol enantiomers in acyl transfer reactions catalysed by Candida antarctica lipase B . Biocatalysis and Biotransformation,16,1-15 .
55. Rotticci, D., Hæffner, F., Orrenius, C., Norin, T., and Hult, K. (1998) Molecular recognition of sec-alcohol enantiomers by Candida antarctica lipase B . Journal of Molecular Catalysis B Enzymatic,5,267-72 .
56. Nakamura, K., Kawasaki, M., and Ohno, A. (1994) Effects of substrate structure on lipase-catalyzed transesterifi cation of ?-substituted 1-alkanols in organic solvents . Bulletin of the Chemical Society of Japan,67,3053-6 .
57. Nakamura, K., Kawasaki, M., and Ohno, A. (1996) Lipase-catalyzed transesterifi cation of aryl-substituted alkanols in an organic solvent . Bulletin of the Chemical Society of Japan,69,1079-85 .
58. Haeffner, F., Norin, T., and Hult, K. (1998) Molecular modeling of the enantioselectivity in lipase-catalyzed transesterifi cation reactions . Biophysical Journal,74,1251-62 .
59. Pineda, J.R.E.T., and Schwartz, S.D. (2006) Protein dynamics and catalysis: the problems of transition state theory and the subtlety of dynamic control . Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences,361,1433-8 .
60. Olsson, M.H.M., Mavri, J., and Warshel, A. (2006) Transition state theory can be used in studies of enzyme catalysis: lessons from simulations of tunnelling and dynamical effects in lipoxygenase and other systems . Philosophical Transactions of the Royal Society of London. Series B Biological Sciences,361,1417-32 .
61. Guieysse, D., Salagnad, C., Monsan, P., Remaud-Simeon, M., and Tran, V. (2003) Towards a novel explanation of Pseudomonas cepacia lipase enantioselectivity via molecular modelling of the enantiomer trajectory into the active site . Tetrahedron: Asymmetry,14,1807-17 .
62. Ozaki, S.-I., and de Montellano, P.R. (1994) Molecular engineering of horseradish peroxidase. Highly enantioselective sulfoxidation of aryl alkyl sulfi des by the Phe-41 ? Leu mutant . Journal of the American Chemical Society,116,4487-8 .
63. Savenkova, M.I., Newmyer, S.L., and de Montellano, P.R. (1996) Rescue of His-42 Ala horseradish peroxidase by a Phe-41 His mutation. Engineering of a surrogate catalytic histidine . The Journal of Biological Chemistry,271,24598-603 .
64. van Loo, B., Lutje Spelberg, J.H., Kingma, J., Sonke, T., Wubbolts, M.G., and Janssen, D.B. (2004) Directed evolution of epoxide hydrolase from A. radiobacter toward higher enantioselectivity by error-prone PCR ad DNA shuffl ing . Chemistry and Biology,11,981-90 .
65. Magnusson, A., Hult, K., and Holmquist, M. (2001) Creation of an enantioselective hydrolase by engineered substrate-assisted catalysis . Journal of the American Chemical Society,123,4354-5 .
66. Reetz, M.T., Wilensek, S., Zha, D., and Jaeger, K.-E. (2001) Directed evolution of an enantioselective enzyme through combinatorial multiple-cassette mutagenesis . Angewandte Chemie-International Edition,40,3589-91 .
67. Park, S., Morley, K.L., Horsman, G.P., Holmquist, M., Hult, K., and Kazlauskas, R.J. (2005) Focusing mutations into the substrate-binding site of Pseudomonas fl uorescens esterase increases the enantioselectivity more effectively thanrandom mutagenesis . Chemistry and Biology,11,45-54 .
68. Henke, E., Bornscheuer, U.T., Schmid, R.D., and Pleiss, J. (2003) A molecular mechanism of enantiorecognition of tertiary alcohols by carboxylesterases . Chembiochem: A European Journal of Chemical Biology,4,485-93 .
69. Ema, T., Fujii, T., Ozaki, M., Korenaga, T., and Sakai, T. (2005) Rational control of enantioselectivity of lipase by site-directed mutagenesis based on the mechanism . Chemical Communications,4650-1 .
70. Rink, R., Lutje-Spelberg, J.H., Pieters, R.J., Kingma, J., Nardini, M., Kellogg, R.M., Dijkstra, B.W., and Janssen, D.B. (1999) Mutation of tyrosine residues involved in the alkylation half reaction of epoxide hydrolase from Agrobacterium radiobacter AD1 results in improved enantioselectivity . Journal of the American Chemical Society,121,7417-18 .
71. Tang, L., van Merode, A.E.J., Lutje Spelberg, J.H., Fraaije, M.W., and Janssen, D.B. (2003) Steady-state kinetics and tryptophan fl uorescence properties of halohydrin dehalogenase from Agrobacterium radiobacter . Roles of W139 and W249 in the active site and halide-induced conformational change . Biochemistry,42,14057-65 .
72. Scheib, H., Pleiss, J., Stadler, P., Kovac, A., Potthoff, A.P., Haalck, L., Spener, F., Paltauf, F., and Schmid, R.D. (1998) Rational design of Rhizopus oryzae lipase with modified stereoselectivity toward triradylglycerols . Protein Engineering,11,675-82 .
73. van Herk, T., Hartog, A.F., Ruijssenaars, H.J., Kerkman, R., Schoemaker, H.E., and Wever, R. (2007) Optimization of the kinetic resolution of the DL-phosphomonoesters of threonine and serine by random mutagenesis of the acid phosphatase from Salmonella enterica . Advanced Synthesis Catalysis,349,1349-52 .
74. Carboni, C., Kierkels, H.G.T., Gardossi, L., Tamiola, K., Janssen, D.B., and Quaedflieg, P.J.L.M. (2006) Preparation of D-amino acids by enzymatic kinetic resolution using a mutant of penicillin-G acylase from E. coli . Tetrahedron: Asymmetry,17,245-51 .
75. Rotticci, D., Rotticci-Mulder, J.C., Denman, S., Norin, T., and Hult, K. (2001) Improved enantioselectivity of a lipase by rational protein engineering . Chembiochem: A European Journal of Chemical Biology,2,766-70 .
76. May, O., Nguyen, P.T., and Arnold, F.H. (2000) Inverting enantioselectivity by directed evolution of hydantoinase for improved production of L-methionine . Nature Biotechnology,18,317-20 .
77. Chen-Goodspeed, M., Sogorb, M.A., Wu, F., and Raushel, F.M. (2001) Structural determinants of the substrate and stereochemical specifi city of phosphotriesterase . Biochemistry,40,1325-31 .
78. Chen-Goodspeed, M., Sogorb, M.A., Wu, F., and Raushel, F.M. (2001) Enhancement, relaxation, and reversal of the stereoselectivity for phosphotriesterase by rational evolution of active site residues . Biochemistry,40,1332-9 .
79. Rui, L., Cao, L., Chen, W., Reardon, K.F., and Wood, T.K. (2005) Protein engineering of epoxide hydrolase from Agrobacterium radiobacter AD1 for enhanced activity and enantioselective production of (R)-1-phenylethane-1,2-diol . Applied and Environmental Microbiology,71,3995-4003 .
80. Lingen, B., Grötzinger, J., Kolter, D., Kula, M.-R., and Pohl, M. (2002) Improving the carboligase activity of benzoylformate decarboxylase from Pseudomonas putida by a combination of directed evolution and site-directed mutagenesis . Protein Engineering,15,585-93 .
81. Kotik, M., Stepanek, V., Kyslik, P., and Maresova, H. (2007) Cloning of an epoxide hydrolase-encoding gene from Aspergillus niger M200, overexpression in E. coli, and modifi cation of activity and enantioselectivity of the enzyme by protein engineering . Journal of Biotechnology,132,8-15 .
82. Manco, G., Carrea, G., Giosue, E., Ottolina, G., Adamo, G., and Rossi, M. (2002) Modifi cation of the enantioselectivity of two homologous thermophilic carboxylesterases from Alicyclobacillus acidocaldarius and Archaeoglobus fulgidus by randommutagenesis and screening . Extremophiles,6,325-31 .
83. Holmquist, M., Martinelle, M., Berglund, P., Clausen, I.G., Patkar, S., Svendsen, A., and Hult, K. (1993) Lipases from Rhizomucor miehei and Humicola lanuginosa : modifi cation of the lid covering the active site alters enantioselectivity . Journal of Protein Chemistry,12,749-57 .
84. Horsman, G.P., Liu, A.M.F., Henke, E., Bornscheuer, U.T., and Kazlauskas, R.J. (2003) Mutations in distant residues moderately increase the enantioselectivity of Pseudomonas fl uorescens esterase toward methyl 3-bromo-2-methylpropanonate and ethyl 3-phenylbutyrate . Chemistry-A European Journal,9,1933-9 .
85. Morley, K.L., and Kazlauskas, R.J. (2005) Improving enzyme properties: when are closer mutations better? Trends in Biotechnology,23,231-7 .
86. Schwede, T., Kopp, J., Guex, N., and Peitsch, M.C. (2003) SWISS-MODEL: an automated protein homology-modeling server . Nucleic Acids Research,31,3381-5 .
87. Clayden, J., Lund, A., Vallverdú, L., and Helliwell, M. (2004) Ultra-remote stereocontrol by conformational communication of information along a carbon chain . Nature,431,966-71 .
88. Lipkowitz, K.B., D'Hue, C.A., Sakamoto, T., and Stack, J.N. (2002) Stereocartography: a computational mapping technique that can locate regions of maximum stereoinduction around chiral catalysts . Journal of the American Chemical Society,124,14255-67 .
89. Parales, R.E., Resnick, S.M., Yu, C.-L., Boyd, D.R., Sharma, N.D., and Gibson, D.T. (2000) Regioselectivity and enantioselectivity of naphthalene dioxygenase during arene cis-dihydroxylation: control by phenylalanine 352 in the a subunit . Journal of Bacteriology,182,5495-504 .
90. Hirose, Y., Kariya, K., Nakanishi, Y., Kurono, Y., and Achiwa, K. (1995) Inversion of enantioselectivity in hydrolysis of 1,4-dihydropyridines by point mutation of lipase PS . Tetrahedron Letters,36,1063-6 .
91. Koga, Y., Kato, K., Nakano, H., and Yamane, T. (2003) Inverting enantioselectivity of Burkholderia cepacia KWI-56 lipase by combinatorial mutagenesis and high-throughput screening using single molecule PCR and in vitro expression . Journal of Molecular Biology,331,585-92 .
92. Ivancic, M., Valinger, G., Gruber, K., and Schwab, H. (2007) Inverting enantioselectivity of Burkholderia gladioliesterase EstB by directed and designed evolution . Journal of Biotechnology,129,109-22 .
93. Bartsch, S., Kourist, R., and Bornscheuer, U.T. (2008) Complete inversion of enantioselectivity towards acetylated tertiary alcohols by a double mutant of a Bacillus subtilis esterase . Angewandte Chemie-International Edition,47,1508-11 .
94. Zha, D., Wilensek, S., Hermes, M., Jaeger, K.-E., and Reetz, M.T. (2001) Complete reversal of enantioselectivity of an enzyme-catalyzed reaction by directed evolution . Chemical Communications,2664-5 .
95. Magnusson, A.O., Takwa, M., Hamberg, A., and Hult, K. (2005) An S-selective lipase was created by rational redesign and the enantioselectivity increased with temperature . Angewandte Chemie-International Edition,44,4582-5 .
96. van den Heuvel, R.H.H., Fraaije, M.W., Ferrer, M., Mattevi, A., and van Berkel, W. J.H. (2000) Inversion of stereospecifi city of vanillyl-alcohol oxidase . Proceedings of the National Academy of Sciences of the United States of America,97,9455-60 .
97. Terao, Y., Ijima, Y., Miyamoto, K., and Ohta, H. (2007) Inversion of enantioselectivity of arylmalonate decarboxylase via site-directed mutation based on the proposed reaction mechanism . Journal of Molecular Catalysis B Enzymatic,45,15-20 .
98. Li, Y., Aubert, S.D., Maes, E.G., and Raushel, F.M. (2004) Enzymatic resolution of chiral phosphinate esters . Journal of the American Chemical Society,126,8888-9 .