Highly efficient "tight fit" immobilization of α-chymotrypsin in mesoporous MCM-41: A novel approach using precursor immobilization and activation

Biotechnology Progress

Volumn 19, Issue 2, 2003, Pages 346-351

  Fadnavis, Nitin W ^a   Bhaskar, Veldurthy ^a   Kantam, Mannepalli Lakshmi ^a   Choudary, Boyapati Manoranjan ^a  

^a INDIAN INSTITUTE OF CHEMICAL TECHNOLOGY   (India)

Author keywords

[No Author keywords available]

Indexed keywords

ENZYME IMMOBILIZATION; MESOPOROUS MATERIALS; PROTEINS; THERMOGRAVIMETRIC ANALYSIS;

REVERSE MICELLAR MEDIA;

BIOTECHNOLOGY;

CHYMOTRYPSIN; CHYMOTRYPSINOGEN; ENZYME ACTIVATION; ENZYME PRECURSORS; ENZYME STABILITY; ENZYMES, IMMOBILIZED; POROSITY; PROTEIN CONFORMATION; SILICON DIOXIDE; TRYPSIN;

EID: 0037360709     PISSN: 87567938     EISSN: None     Source Type: Journal    
DOI: 10.1021/bp025678s     Document Type: Article

References (40)

1. Drauz, K.; Waldmann, H.; Sauerbrei, B. Applied Homogeneous Catalysis with Organometallic Compounds: A Comprehensive Handbook in Three Volumes; Cornils, B., Herrmann, W. A., Eds.; VCH: Weinheim, 1996; Vol. 2, pp 768-801. Schmid, R. D.; Verger, R. Lipases: Interfacial enzymes with attractive applications. Angew. Chem., Int. Ed. 1998, 37, 1608-1633. Gill, I.; Pastor, E.; Ballesteros, A. Lipase-silicone biocomposites: Efficient and versatile immobilized biocatalysts. J. Am. Chem. Soc. 1999, 121, 9487-9496.
2. Drauz, K.; Waldmann, H.; Sauerbrei, B. Applied Homogeneous Catalysis with Organometallic Compounds: A Comprehensive Handbook in Three Volumes; Cornils, B., Herrmann, W. A., Eds.; VCH: Weinheim, 1996; Vol. 2, pp 768-801. Schmid, R. D.; Verger, R. Lipases: Interfacial enzymes with attractive applications. Angew. Chem., Int. Ed. 1998, 37, 1608-1633. Gill, I.; Pastor, E.; Ballesteros, A. Lipase-silicone biocomposites: Efficient and versatile immobilized biocatalysts. J. Am. Chem. Soc. 1999, 121, 9487-9496.
3. Drauz, K.; Waldmann, H.; Sauerbrei, B. Applied Homogeneous Catalysis with Organometallic Compounds: A Comprehensive Handbook in Three Volumes; Cornils, B., Herrmann, W. A., Eds.; VCH: Weinheim, 1996; Vol. 2, pp 768-801. Schmid, R. D.; Verger, R. Lipases: Interfacial enzymes with attractive applications. Angew. Chem., Int. Ed. 1998, 37, 1608-1633. Gill, I.; Pastor, E.; Ballesteros, A. Lipase-silicone biocomposites: Efficient and versatile immobilized biocatalysts. J. Am. Chem. Soc. 1999, 121, 9487-9496.
4. Morihara, K.; Oka, T. α-Chymotrypsin as the catalyst for peptide synthesis. Biochem. J. 1977, 163, 531-542. West, B. J.; Scholten, J.; Stolowich, N. J.; Hogg, J.; Scott, A. I.; Wong, C.-H. Modification of proteases to esterases for peptide synthesis: Methylchymotrypsin. J. Am. Chem. Soc. 1988, 110, 3709-3712. Miyazawa, T.; Tanaka, K.; Eusantsu, E.; Yanagihara, E.; Yamada, T. Remarkable effects of donor esters on the α-chymotrypsin-catalyzed coupling of inherently poor amino acid substrates. Tetrahedron Lett. 1998, 39, 997-1000. Oaki, J.; Nakahara, K.; Tamura, M.; Okai, H. One step synthesis of inverted aspartame type sweetener, Ac-Phe-Lys, using chemically modified chymotrypsin. Biosci. Biotechnol. Biochem. 1999, 63, 1156-1159. Luthi, P.; Luisi, P. L. Enzymatic synthesis of hydrocarbon soluble peptides with reverse micelles. J. Am. Chem. Soc. 1984, 106, 7285-7286. Fadnavis, N. W.; Luisi, P. L. Immobilized enzymes in reverse micelles: Studies with gel-entrapped trypsin and α-chymotrypsin in AOT reverse micelles. Biotechnol. Bioeng. 1989, 33, 1277-1282.
5. Morihara, K.; Oka, T. α-Chymotrypsin as the catalyst for peptide synthesis. Biochem. J. 1977, 163, 531-542. West, B. J.; Scholten, J.; Stolowich, N. J.; Hogg, J.; Scott, A. I.; Wong, C.-H. Modification of proteases to esterases for peptide synthesis: Methylchymotrypsin. J. Am. Chem. Soc. 1988, 110, 3709-3712. Miyazawa, T.; Tanaka, K.; Eusantsu, E.; Yanagihara, E.; Yamada, T. Remarkable effects of donor esters on the α-chymotrypsin-catalyzed coupling of inherently poor amino acid substrates. Tetrahedron Lett. 1998, 39, 997-1000. Oaki, J.; Nakahara, K.; Tamura, M.; Okai, H. One step synthesis of inverted aspartame type sweetener, Ac-Phe-Lys, using chemically modified chymotrypsin. Biosci. Biotechnol. Biochem. 1999, 63, 1156-1159. Luthi, P.; Luisi, P. L. Enzymatic synthesis of hydrocarbon soluble peptides with reverse micelles. J. Am. Chem. Soc. 1984, 106, 7285-7286. Fadnavis, N. W.; Luisi, P. L. Immobilized enzymes in reverse micelles: Studies with gel-entrapped trypsin and α-chymotrypsin in AOT reverse micelles. Biotechnol. Bioeng. 1989, 33, 1277-1282.
6. Morihara, K.; Oka, T. α-Chymotrypsin as the catalyst for peptide synthesis. Biochem. J. 1977, 163, 531-542. West, B. J.; Scholten, J.; Stolowich, N. J.; Hogg, J.; Scott, A. I.; Wong, C.-H. Modification of proteases to esterases for peptide synthesis: Methylchymotrypsin. J. Am. Chem. Soc. 1988, 110, 3709-3712. Miyazawa, T.; Tanaka, K.; Eusantsu, E.; Yanagihara, E.; Yamada, T. Remarkable effects of donor esters on the α-chymotrypsin-catalyzed coupling of inherently poor amino acid substrates. Tetrahedron Lett. 1998, 39, 997-1000. Oaki, J.; Nakahara, K.; Tamura, M.; Okai, H. One step synthesis of inverted aspartame type sweetener, Ac-Phe-Lys, using chemically modified chymotrypsin. Biosci. Biotechnol. Biochem. 1999, 63, 1156-1159. Luthi, P.; Luisi, P. L. Enzymatic synthesis of hydrocarbon soluble peptides with reverse micelles. J. Am. Chem. Soc. 1984, 106, 7285-7286. Fadnavis, N. W.; Luisi, P. L. Immobilized enzymes in reverse micelles: Studies with gel-entrapped trypsin and α-chymotrypsin in AOT reverse micelles. Biotechnol. Bioeng. 1989, 33, 1277-1282.
7. Morihara, K.; Oka, T. α-Chymotrypsin as the catalyst for peptide synthesis. Biochem. J. 1977, 163, 531-542. West, B. J.; Scholten, J.; Stolowich, N. J.; Hogg, J.; Scott, A. I.; Wong, C.-H. Modification of proteases to esterases for peptide synthesis: Methylchymotrypsin. J. Am. Chem. Soc. 1988, 110, 3709-3712. Miyazawa, T.; Tanaka, K.; Eusantsu, E.; Yanagihara, E.; Yamada, T. Remarkable effects of donor esters on the α-chymotrypsin-catalyzed coupling of inherently poor amino acid substrates. Tetrahedron Lett. 1998, 39, 997-1000. Oaki, J.; Nakahara, K.; Tamura, M.; Okai, H. One step synthesis of inverted aspartame type sweetener, Ac-Phe-Lys, using chemically modified chymotrypsin. Biosci. Biotechnol. Biochem. 1999, 63, 1156-1159. Luthi, P.; Luisi, P. L. Enzymatic synthesis of hydrocarbon soluble peptides with reverse micelles. J. Am. Chem. Soc. 1984, 106, 7285-7286. Fadnavis, N. W.; Luisi, P. L. Immobilized enzymes in reverse micelles: Studies with gel-entrapped trypsin and α-chymotrypsin in AOT reverse micelles. Biotechnol. Bioeng. 1989, 33, 1277-1282.
8. Morihara, K.; Oka, T. α-Chymotrypsin as the catalyst for peptide synthesis. Biochem. J. 1977, 163, 531-542. West, B. J.; Scholten, J.; Stolowich, N. J.; Hogg, J.; Scott, A. I.; Wong, C.-H. Modification of proteases to esterases for peptide synthesis: Methylchymotrypsin. J. Am. Chem. Soc. 1988, 110, 3709-3712. Miyazawa, T.; Tanaka, K.; Eusantsu, E.; Yanagihara, E.; Yamada, T. Remarkable effects of donor esters on the α-chymotrypsin-catalyzed coupling of inherently poor amino acid substrates. Tetrahedron Lett. 1998, 39, 997-1000. Oaki, J.; Nakahara, K.; Tamura, M.; Okai, H. One step synthesis of inverted aspartame type sweetener, Ac-Phe-Lys, using chemically modified chymotrypsin. Biosci. Biotechnol. Biochem. 1999, 63, 1156-1159. Luthi, P.; Luisi, P. L. Enzymatic synthesis of hydrocarbon soluble peptides with reverse micelles. J. Am. Chem. Soc. 1984, 106, 7285-7286. Fadnavis, N. W.; Luisi, P. L. Immobilized enzymes in reverse micelles: Studies with gel-entrapped trypsin and α-chymotrypsin in AOT reverse micelles. Biotechnol. Bioeng. 1989, 33, 1277-1282.
9. Morihara, K.; Oka, T. α-Chymotrypsin as the catalyst for peptide synthesis. Biochem. J. 1977, 163, 531-542. West, B. J.; Scholten, J.; Stolowich, N. J.; Hogg, J.; Scott, A. I.; Wong, C.-H. Modification of proteases to esterases for peptide synthesis: Methylchymotrypsin. J. Am. Chem. Soc. 1988, 110, 3709-3712. Miyazawa, T.; Tanaka, K.; Eusantsu, E.; Yanagihara, E.; Yamada, T. Remarkable effects of donor esters on the α-chymotrypsin-catalyzed coupling of inherently poor amino acid substrates. Tetrahedron Lett. 1998, 39, 997-1000. Oaki, J.; Nakahara, K.; Tamura, M.; Okai, H. One step synthesis of inverted aspartame type sweetener, Ac-Phe-Lys, using chemically modified chymotrypsin. Biosci. Biotechnol. Biochem. 1999, 63, 1156-1159. Luthi, P.; Luisi, P. L. Enzymatic synthesis of hydrocarbon soluble peptides with reverse micelles. J. Am. Chem. Soc. 1984, 106, 7285-7286. Fadnavis, N. W.; Luisi, P. L. Immobilized enzymes in reverse micelles: Studies with gel-entrapped trypsin and α-chymotrypsin in AOT reverse micelles. Biotechnol. Bioeng. 1989, 33, 1277-1282.
10. Santaniello, E.; Chiari, M.; Ferraboschi, P.; Trave, S. Enhanced and reversed enantioselectivity of enzymatic hydrolysis by simple substrate modifications: The case of 3-hydroxyglutarate diesters. J. Org. Chem. 1988, 53, 1567-1569. Schricker, B.; Thirring, K.; Berner, H. α-Chymotrypsin catalysed enantioselective hydrolysis of alkenyl-α-amino acid esters. Biorg. Med. Chem. Lett. 1992, 2, 387-390. Ricca, J.-M.; Crout, D. H. G. Selectivity and specificity in substrate binding to proteases: Novel hydrolytic reactions catalysed by α-chymotrypsin suspended in organic solvents with low water content and mediated by ammonium hydrogen carbonate. J. Chem. Soc., Perkin Trans. 1993, 1, 1225-1233. Mohr, P.; Rosslein, L.; Tamm, C. Hydroxyglutarate and β,γ-epoxy esters as substrates for pig liver esterase (PLE) and α-chymotrypsin. Helv. Chim. Acta 1987, 70, 142-152.
11. Santaniello, E.; Chiari, M.; Ferraboschi, P.; Trave, S. Enhanced and reversed enantioselectivity of enzymatic hydrolysis by simple substrate modifications: The case of 3-hydroxyglutarate diesters. J. Org. Chem. 1988, 53, 1567-1569. Schricker, B.; Thirring, K.; Berner, H. α-Chymotrypsin catalysed enantioselective hydrolysis of alkenyl-α-amino acid esters. Biorg. Med. Chem. Lett. 1992, 2, 387-390. Ricca, J.-M.; Crout, D. H. G. Selectivity and specificity in substrate binding to proteases: Novel hydrolytic reactions catalysed by α-chymotrypsin suspended in organic solvents with low water content and mediated by ammonium hydrogen carbonate. J. Chem. Soc., Perkin Trans. 1993, 1, 1225-1233. Mohr, P.; Rosslein, L.; Tamm, C. Hydroxyglutarate and β,γ-epoxy esters as substrates for pig liver esterase (PLE) and α-chymotrypsin. Helv. Chim. Acta 1987, 70, 142-152.
12. Santaniello, E.; Chiari, M.; Ferraboschi, P.; Trave, S. Enhanced and reversed enantioselectivity of enzymatic hydrolysis by simple substrate modifications: The case of 3-hydroxyglutarate diesters. J. Org. Chem. 1988, 53, 1567-1569. Schricker, B.; Thirring, K.; Berner, H. α-Chymotrypsin catalysed enantioselective hydrolysis of alkenyl-α-amino acid esters. Biorg. Med. Chem. Lett. 1992, 2, 387-390. Ricca, J.-M.; Crout, D. H. G. Selectivity and specificity in substrate binding to proteases: Novel hydrolytic reactions catalysed by α-chymotrypsin suspended in organic solvents with low water content and mediated by ammonium hydrogen carbonate. J. Chem. Soc., Perkin Trans. 1993, 1, 1225-1233. Mohr, P.; Rosslein, L.; Tamm, C. Hydroxyglutarate and β,γ-epoxy esters as substrates for pig liver esterase (PLE) and α-chymotrypsin. Helv. Chim. Acta 1987, 70, 142-152.
13. Santaniello, E.; Chiari, M.; Ferraboschi, P.; Trave, S. Enhanced and reversed enantioselectivity of enzymatic hydrolysis by simple substrate modifications: The case of 3-hydroxyglutarate diesters. J. Org. Chem. 1988, 53, 1567-1569. Schricker, B.; Thirring, K.; Berner, H. α-Chymotrypsin catalysed enantioselective hydrolysis of alkenyl-α-amino acid esters. Biorg. Med. Chem. Lett. 1992, 2, 387-390. Ricca, J.-M.; Crout, D. H. G. Selectivity and specificity in substrate binding to proteases: Novel hydrolytic reactions catalysed by α-chymotrypsin suspended in organic solvents with low water content and mediated by ammonium hydrogen carbonate. J. Chem. Soc., Perkin Trans. 1993, 1, 1225-1233. Mohr, P.; Rosslein, L.; Tamm, C. Hydroxyglutarate and β,γ-epoxy esters as substrates for pig liver esterase (PLE) and α-chymotrypsin. Helv. Chim. Acta 1987, 70, 142-152.
14. Ward, T. J. Chiral separation. Anal. Chem. 2000, 72, 4521-4528.
15. Reslow, M.; Adlercreutz, P.; Mattiasson, B. On the importance of the support material for bioorganic synthesis, Influence of water partition between solvent, enzyme and solid support in water poor reaction media. Eur. J. Biochem. 1988, 172, 573-578.
16. Tuel, A.; Gontier, S. Synthesis and characterization of trivalent metal containing mesoporous silica obtained by a natural templating route. Chem. Mater. 1996, 8, 114-122; Chem. Abstr. 1996, 124, 44015s.
17. Tuel, A.; Gontier, S. Synthesis and characterization of trivalent metal containing mesoporous silica obtained by a natural templating route. Chem. Mater. 1996, 8, 114-122; Chem. Abstr. 1996, 124, 44015s.
18. Fadnavis, N. W.; Chandraprakash, Y.; Deshpande, A. Protein-protein interactions in reverse micelles: Trypsin shows superactivity towards a protein substrate α-chymotrypsinogen A in reverse micelles of sodium bis (2-ethylhexyl)-sulfosuccinate (AOT) in isooctane. Biochemie 1993, 75, 995-999. Kantrowitz, E. R.; Eisele, G. Two spectrophotometic experiments with α-cymotrypsin. J. Chem. Educ. 1975, 52, 410-413.
19. Fadnavis, N. W.; Chandraprakash, Y.; Deshpande, A. Protein-protein interactions in reverse micelles: Trypsin shows superactivity towards a protein substrate α-chymotrypsinogen A in reverse micelles of sodium bis (2-ethylhexyl)-sulfosuccinate (AOT) in isooctane. Biochemie 1993, 75, 995-999. Kantrowitz, E. R.; Eisele, G. Two spectrophotometic experiments with α-cymotrypsin. J. Chem. Educ. 1975, 52, 410-413.
20. Laszlo, P.; Mathy, A. Catalysis of Friedal-Crafts alkylation by a montmorillonite doped with transition metal cations. Helv. Chim. Acta 1987, 70, 577-586.
21. Hammond. B. R.; Gutfreund. H. Two steps in the reaction of chymotrypsin with acetyl-L-phenylalanine ethyl ester. Biochem. J. 1955, 61, 187-189.
22. Gentile, A.; Giordano, C.; Fuganti, C.; Ghirotto, L.; Servi, S. The enzymatic preparation of (2R,3S)-phenyl glycidic acid esters. J. Org. Chem. 1992, 57, 6635-6637.
23. Matthews, B. M.; Sigler, P. D.; Henderson, H.; Blow, D. M. Three-dimensional structure of tosyl-α-chymotrypsin. Nature 1967, 214, 652-656. Squire. P. G.; Himmel, M. E. Hydrodynamics and protein hydration. Arch. Biochem. Biophys. 1979, 196, 165-177. Levashov, A. V.; Khmelnitsky, Y. L.; Klyachko, N. L.; Chernyak, V.; Martinek, K. Enzymes entrapped into reverse micelles in organic solvents. J. Colloid Interface Sci. 1982, 88, 444-457.
24. Matthews, B. M.; Sigler, P. D.; Henderson, H.; Blow, D. M. Three-dimensional structure of tosyl-α-chymotrypsin. Nature 1967, 214, 652-656. Squire. P. G.; Himmel, M. E. Hydrodynamics and protein hydration. Arch. Biochem. Biophys. 1979, 196, 165-177. Levashov, A. V.; Khmelnitsky, Y. L.; Klyachko, N. L.; Chernyak, V.; Martinek, K. Enzymes entrapped into reverse micelles in organic solvents. J. Colloid Interface Sci. 1982, 88, 444-457.
25. Matthews, B. M.; Sigler, P. D.; Henderson, H.; Blow, D. M. Three-dimensional structure of tosyl-α-chymotrypsin. Nature 1967, 214, 652-656. Squire. P. G.; Himmel, M. E. Hydrodynamics and protein hydration. Arch. Biochem. Biophys. 1979, 196, 165-177. Levashov, A. V.; Khmelnitsky, Y. L.; Klyachko, N. L.; Chernyak, V.; Martinek, K. Enzymes entrapped into reverse micelles in organic solvents. J. Colloid Interface Sci. 1982, 88, 444-457.
26. Beck, J. S.; Vartuli, J. C.; Roth, W. J.; Leonowiez, M. E.; Kresge, C. T.; Schmitt, C. D.; Chu, C. T. W.; Olson, D. H.; Sheppard, E. W.; McCullen, S. B.; Higgins, J. B.; Schlenker, J. L. A new family of mesoporous molecular seives prepared with liquid crystal templates. J. Am. Chem. Soc. 1992, 114, 10834-10854. Kresge, C. T.; Leonowiez, M. E.; Roth, W. J.; Vartuli, J. C.; Beck., J. S. Ordered mesoporous molecular sieves synthesized by a liquid-crystal mechanism. Nature 1992, 359, 710-712.
27. Beck, J. S.; Vartuli, J. C.; Roth, W. J.; Leonowiez, M. E.; Kresge, C. T.; Schmitt, C. D.; Chu, C. T. W.; Olson, D. H.; Sheppard, E. W.; McCullen, S. B.; Higgins, J. B.; Schlenker, J. L. A new family of mesoporous molecular seives prepared with liquid crystal templates. J. Am. Chem. Soc. 1992, 114, 10834-10854. Kresge, C. T.; Leonowiez, M. E.; Roth, W. J.; Vartuli, J. C.; Beck., J. S. Ordered mesoporous molecular sieves synthesized by a liquid-crystal mechanism. Nature 1992, 359, 710-712.
28. Zerner, B.; Bond, R. P. M.; Bender, M. L. Kinetic evidence for the formation of acyl-enzyme intermediates in the α-chymotrypsin-catalyzed hydrolysis of specific substrates. J. Am. Chem. Soc. 1964, 86, 3674-3679.
29. Sreenivas Rao, A.; Bhalerao, U. T.; Fadnavis, N. W. A novel tea-bag methodology for enzymatic resolutions of α-amino acid derivatives in reverse micellar media. Synth. Commun. 1994, 24, 2109-2118.
30. Elks, J.; Ganellin, C. R. Dictionary of Drugs; Champman and Hall: London, 1990; p 426. Kanerva, L. T.; Smdholm, O. Lipase catalysis in the resolution of recemic intermediates of diltiazem synthesis in organic solvents. J. Chem. Soc., Perkin Trans. 1 1993, 1385-1389.
31. Elks, J.; Ganellin, C. R. Dictionary of Drugs; Champman and Hall: London, 1990; p 426. Kanerva, L. T.; Smdholm, O. Lipase catalysis in the resolution of recemic intermediates of diltiazem synthesis in organic solvents. J. Chem. Soc., Perkin Trans. 1 1993, 1385-1389.
32. Deng, L.; Jocobsen, E. N. A practical, highly enantioselective synthesis of the Taxol side chain via asymmetric catalysis. J. Org. Chem. 1992, 57, 4320-4323. Honig. H.; Senfer, P.; Wasserthal, H. W. Chemo-enzymatic synthesis of all isomeric 3-phenylseranes and -isoseranes. Tetrahedron 1990, 46, 3841. Gou, D. M.; Liu. Y. C.; Chen, C. S. A practical chemoselective synthesis of the Taxol C-13 side chain N-benzoyl-(2R,3S)-3-phenylisoserine. J. Org. Chem. 1993, 58, 1287-1289.
33. Deng, L.; Jocobsen, E. N. A practical, highly enantioselective synthesis of the Taxol side chain via asymmetric catalysis. J. Org. Chem. 1992, 57, 4320-4323. Honig. H.; Senfer, P.; Wasserthal, H. W. Chemo-enzymatic synthesis of all isomeric 3-phenylseranes and -isoseranes. Tetrahedron 1990, 46, 3841. Gou, D. M.; Liu. Y. C.; Chen, C. S. A practical chemoselective synthesis of the Taxol C-13 side chain N-benzoyl-(2R,3S)-3-phenylisoserine. J. Org. Chem. 1993, 58, 1287-1289.
34. Deng, L.; Jocobsen, E. N. A practical, highly enantioselective synthesis of the Taxol side chain via asymmetric catalysis. J. Org. Chem. 1992, 57, 4320-4323. Honig. H.; Senfer, P.; Wasserthal, H. W. Chemo-enzymatic synthesis of all isomeric 3-phenylseranes and -isoseranes. Tetrahedron 1990, 46, 3841. Gou, D. M.; Liu. Y. C.; Chen, C. S. A practical chemoselective synthesis of the Taxol C-13 side chain N-benzoyl-(2R,3S)-3-phenylisoserine. J. Org. Chem. 1993, 58, 1287-1289.
35. Lopez, J. L.; Matson, S. L. A multiphase/extractive enzyme membrane reactor for production of Diltiazem chiral intermediate. J. Membr. Sci. 1997, 125, 189-211.
36. Chen, C. S.; Fujimoto, Y.; Girdokas, G.; Sih, E. Quantitative analysis of biochemical kinetic resolutions of enantiomers. J. Am. Chem. Soc. 1982, 104, 7294-7299.
37. Luisi, P. L.; Magid, L. J. Solubilization of enzymes and nucleic acids in hydrocarbon micellar solutions. CRC Crit. Rev. Biochem. 1986, 20, 409-474.
38. Partridge, J.; Moore, B. D.; Halling, P. J. α-Chymotrypsin stability in aqueous-acetonitrile mixtures: is the native enzyme thermodynamically or kinetically stable under low water conditions? J. Mol. Catal. B 1999, 6, 11-20.
39. Diaz, J. F.; Balkus, K. J., Jr. Enzyme immobilization in MCM-41 molecular sieve. J. Mol. Catal. B 1996, 2, 115-126.
40. Baron, M. H.; Revault, M.; Noinville, S. S.; Abadie, J.; Quiquampoix, H. Chymotrypsin adsorption on montmorillonite: enzyme activity and kinetic FT-IR structural analysis. J. Colloid Interface Sci. 1999, 214, 319-332.