Tweel / Harder / Buitelaar Stability and Stabilization of Enzymes

Proceedings of an International Symposium Held in Maastricht, The Netherlands, 22-25 November 1992
1. Auflage 2013
ISBN: 978-1-4832-9133-8
Verlag: Elsevier Science & Techn.
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band Volume 47, 535 Seiten, Web PDF

Reihe: Studies in Organic Chemistry

These proceedings contain most of the oral presentations and posters of the international symposium on Stability and Stabilization of Enzymes held in Maastricht in November 1992. They provide a comprehensive overview of the state-of-the-art in this field.The possible applications of enzymes are enormous. Years of development have seen many enzymes brought onto the market, but they are still expensive to use. Therefore, their efficient application is a prerequisite for common usage. One of the main factors for this efficiency is the stability of the enzymes. The topics thus ranged from the extensive fundamental thermodynamic knowledge gathered in academic research to the practical applied knowledge built up in industry during the time that enzymes have been produced commercially.The subject Stability and Stabilization of Enzymes was discussed from various points of view, as was reflected in the themes of the symposium sessions. In the session on Fundamentals of Enzyme Stabilisation the thermodynamic background of the phenomenon was highlighted. In yet another session, the recently developed analytical tools to measure enzyme stability and stabilisation were discussed. Further sessions comprised the physical, chemical and biological ways to obtain enzyme stabilisation and finally, the industrial practice of enzyme stabilisation was treated by representatives of the world's most important enzyme producers. The book will be of interest to researchers in universities and industry in the fields of biochemistry, enzymology and biotechnology.

Autoren/Hrsg.

Tweel, W. J. J. van den

Harder, A.

Buitelaar, R. M.

Inhaltsverzeichnis

1;Front Cover;1
2;Stability and Stabilization of Enzymes;4
3;Copyright Page;5
4;Table of Contents;10
5;Studies in Organic Chemistry;6
6;PREFACE;8
7;ACKNOWLEDGEMENTS;9
8;PART 1: ORAL PRESENTATIONS;16
8.1;Chapter 1. The behaviour of proteins at interfaces in relation to their structural stability;18
8.1.1;Abstract;18
8.1.2;1. INTRODUCTION;18
8.1.3;2. TYPES OF INTERACTION THAT DETERMINE PROTEIN ADSORPTION;19
8.1.4;3. CASE STUDIES: PROTEIN ADSORPTION IN MODEL SYSTEMS;22
8.1.5;6. REFERENCES;25
8.2;Chapter 2. Influence of the solvent properties on protein stability in organic media;28
8.2.1;Abstract;28
8.2.2;1. INTRODUCTION;28
8.2.3;2. MATERIALS AND METHODS;29
8.2.4;3. RESULTS AND DISCUSSION;30
8.2.5;4. REFERENCES;35
8.3;Chapter 3. Interfacial damage to proteins during intensive mixing in fermentation and downstream processing;36
8.3.1;Abstract;36
8.3.2;1. INTRODUCTION;36
8.3.3;2. PUMPS;37
8.3.4;3. ULTRAFILTERS;37
8.3.5;4. MIXERS, REACTORS, FERMENTERS;39
8.3.6;5. CONCLUSIONS;40
8.3.7;6. REFERENCES;41
8.3.8;7. ACKNOWLEDGEMENT;42
8.4;Chapter 4. Thermal stability of enzymes: influence of solvatation medium (a Raman spectroscopy study);44
8.4.1;Abstract;44
8.4.2;1. INTRODUCTION;44
8.4.3;2. SOLVENT - ADDITIVES INTERACTIONS;45
8.4.4;3. CONFORMATIONAL CHANGES OF ENZYMES;45
8.4.5;5. CONCLUSION;50
8.4.6;6. REFERENCES;51
8.5;CHAPTER 5. PROTEIN STABILITY IN NON-AQUEOUS MEDIA: A DSC STUDY;52
8.5.1;Abstract;52
8.5.2;1. INTRODUCTION;52
8.5.3;2. EXPERIMENTAL;53
8.5.4;3.RESULTS;54
8.5.5;4. MODEL SYSTEMS;56
8.5.6;5. DISCUSSION;57
8.5.7;6. REFERENCES;59
8.6;Chapter 6. Stable enzymes by water removal;60
8.6.1;Abstract;60
8.6.2;l. INTRODUCTION;60
8.6.3;2. ICE NUCLEATION AND CRYSTAL GROWTH;61
8.6.4;3. CHEMISTRY AND BIOCHEMISTRY OF FREEZING;62
8.6.5;4. Supersaturation and Vitrification: The Role of Excipients;63
8.6.6;5. VERY CONCENTRATED MIXTURES: WATER AS PLASTICIZER;65
8.6.7;6. PROCESS ANALYSIS OF FREEZE DRYING;65
8.6.8;7. CHOICE OF FORMULATION;66
8.6.9;8. STORAGE STABILITY;67
8.6.10;9. GLASSY STATE STABILIZATION BY EVAPORATION;67
8.6.11;9. CONCLUSIONS;69
8.6.12;10. REFERENCES;69
8.7;CHAPTER 7. ENZYME STABILIZATION BY MULTIPOINT COVALENT ATTACHMENT TO ACTIVATED PRE-EXISTING SUPPORTS;70
8.7.1;INTRODUCTION;70
8.7.2;1. THE IMMOBILIZATION/STABILIZATION SYSTEM;71
8.7.3;2. ACTIVATION OF SUPPORTS AND ENZYME IMMOBILIZATION;72
8.7.4;3. ONE-POINT IMMOBILIZATION;73
8.7.5;4. MULTI-POINT IMMOBILIZATION. THE STRATEGY OF STABILIZATION;73
8.7.6;5. GENERAL APPLICABILITY OF OUR STABILIZATION STRATEGY;75
8.7.7;6. MULTIPOINT IMMOBILIZATION AS A STARTING POINT TO ADDITIONAL STABILIZATION & REACTIVATION STRATEGIES;75
8.7.8;7. CONCLUDING REMARKS;76
8.7.9;8. REFERENCES;77
8.8;Chapter 8. Crosslinked enzyme crystals (CLECs™) as immobilized enzyme particles;78
8.8.1;Abstract;78
8.8.2;1. INTRODUCTION;78
8.8.3;2. CRYSTALLIZATION OF PROTEINS;79
8.8.4;3. MECHANICAL STABILITY OF CRYSTALS;81
8.8.5;4. THE PROBLEM OF DIFFUSION;84
8.8.6;5. CONCLUSION;86
8.8.7;6. REFERENCES;87
8.9;Chapter 9. Stabilization of proteins by chemical methods;90
8.9.1;Abstract;90
8.9.2;DIFFERENT TYPES OF PROTEIN STABILITY;90
8.9.3;MECHANISMS OF IRREVERSIBLE THERMOINACTIVATION;91
8.9.4;IDEAS ON HOW TO INCREASE LONG-TERM STABILITY;91
8.9.5;IMMOBILIZATION EFFICIENTLY INHIBITS PROTEIN UNFOLDING;93
8.9.6;STABILIZATION AGAINST "INCORRECT REFOLDING";93
8.9.7;POSSIBLE LIMIT IN THERMOSTABILITY OF ENZYMES;94
8.9.8;REFERENCES;94
8.10;Chapter 10. Stability of engineered antibody fragments;96
8.10.1;Abstract;96
8.10.2;1. INTRODUCTION;96
8.10.3;2. ANTIBODY STRUCTURE;97
8.10.4;3. DOMAIN DISSOCIATION AND IRREVERSIBLE DENATURATION;99
8.10.5;4 UNFOLDING OF INDIVIDUAL DOMAINS;104
8.10.6;5. ACKNOWLEDGEMENTS;104
8.10.7;6. REFERENCES;105
8.11;CHAPTER 11. STRUCTURAL DETERMINANTS OF THE THERMOSTABILITY OF THERMOLYSIN-LIKE BACILLUS NEUTRAL PROTEASES;106
8.11.1;Abstract;106
8.11.2;1. INTRODUCTION;106
8.11.3;2. MATERIALS AND METHODS;108
8.11.4;3. RESULTS;109
8.11.5;4. DISCUSSION;112
8.11.6;5. REFERENCES;113
8.12;CHAPTER 12. MOLECULAR ASPECTS OF PROTEOLYSIS OF GLOBULAR PROTEINS;116
8.12.1;Abstract;116
8.12.2;1. INTRODUCTION;116
8.12.3;2. EXPOSED AND FLEXIBLE LOOPS ARE THE SITES OF LIMITED PROTEOLYSIS;116
8.12.4;3. DISCUSSION;121
8.12.5;Acknowledgments;123
8.12.6;4. REFERENCES;123
8.13;CHAPTER 13. STABILITY OF INDUSTRIAL ENZYMES;126
8.13.1;Abstract;126
8.13.2;1. INTRODUCTION;127
8.13.3;2. FACTORS AFFECTING THE STABILITY OF ENZYMES;131
8.13.4;3. STABILISATION OF INDUSTRIAL ENZYMES;132
8.13.5;4. EXAMPLES;137
8.13.6;5. ACKNOWLEDGMENT;146
8.13.7;6. REFERENCES;146
8.14;Chapter 14. Stabilization of the Detergent Protease Savinase® by Proline Substitution;148
8.14.1;Abstract;148
8.14.2;1. INTRODUCTION;148
8.14.3;2. MATERIALS AND METHODS;149
8.14.4;3. RESULTS AND DISCUSSION;151
8.14.5;4. CONCLUSIONS;157
8.14.6;5. ACKNOWLEDGEMENTS;157
8.14.7;6. REFERENCES;157
8.15;Chapter 15. Lessons from Industry;160
8.15.1;1. INTRODUCTION;160
8.15.2;2. GENERAL CRITERIA FOR AN INDUSTRIAL BIOCATALYST;161
8.15.3;3. EXAMPLES OF INDUSTRIALLY IMPORTANT ENZYMES;162
8.15.4;4. REFERENCES;166
8.16;CHAPTER 16. STABILIZATION OF ENZYMES BY THEIR SPECIFIC ANTIBODIES;168
8.16.1;1. INTRODUCTION;168
8.16.2;2. METHODS;169
8.16.3;3. RESULTS AND DISCUSSION;172
8.16.4;4. GENERAL FEATURES;176
8.16.5;5. REFERENCES;177
9;PART 2: POSTERS;180
9.1;Chapter 17. Immobilisation of a-Chymotrypsin on soluble acrylic microgels; activity and stabilisation;182
9.1.1;l. INTRODUCTION;182
9.1.2;2. EXPERIMENTAL;183
9.1.3;3. RESULTS AND DISCUSSION;187
9.1.4;REFERENCES;189
9.2;Chapter 18. Improved Stability of Lignin Peroxidase by Immobilization;190
9.2.1;1. INTRODUCTION;190
9.2.2;2. MATERIAL AND METHODS;190
9.2.3;3. RESULTS AND DISCUSSION;191
9.2.4;CONCLUSION;194
9.2.5;REFERENCES;195
9.3;Chapter 19. Ca2+-induced Enhancement of the Molecular Stability of Pseudomonas Lipases;196
9.3.1;1. INTRODUCTION;196
9.3.2;2. MATERIALS AND METHODS;197
9.3.3;3. RESULTS AND DISCUSSIONS;198
9.3.4;4. REFERENCES;203
9.4;CHAPTER 20. DENATURATION OF RIBONUCLEASES FROM DIFFERENT SOURCES IN THE PRESENCE OF DENATURING OR STABILIZING AGENTS;204
9.4.1;1. THE RIBONUCLEASES;204
9.4.2;2. EXPERIMENTAL;205
9.4.3;3. DSC RESULTS;205
9.4.4;4. DENATURATION OF RNAase A IN THE PRESENCE OF DIFFERENT COSOLUTES;210
9.4.5;5. REFERENCES;211
9.5;Chapter 21. Conformational dynamics of native, compact and fully unfolded states of proteins detected by frequency domain fluorometry;212
9.5.1;1. INTRODUCTION;212
9.5.2;2. EXPERIMENTAL SECTION;213
9.5.3;3. RESULTS;214
9.5.4;4. DISCUSSION;218
9.5.5;5. REFERENCES;218
9.6;Chapter 22. Characteristics, protein engineering and applications of psychrophilic marine proteinases from Atlantic cod;220
9.6.1;1. INTRODUCTION;221
9.6.2;2. TRYPSIN FROM ATLANTIC COD;222
9.6.3;3. CHYMOTRYPSIN FROM ATLANTIC COD;223
9.6.4;4. ELASTASE FROM ATLANTIC COD;224
9.6.5;5. COLLAGENASE FROM ATLANTIC COD;226
9.6.6;6. CONCLUSIONS;227
9.6.7;7. REFERENCES;228
9.7;CHAPTER 23. LOW TEMPERATURE INACTIVATION OF A BACTERIAL PROTEASE;230
9.7.1;1. INTRODUCTION;230
9.7.2;2. MATERIAL AND METHODS;231
9.7.3;3. RESULTS AND DISCUSSION;232
9.7.4;4. ACKNOWLEDGEMENTS;237
9.7.5;5. REFERENCES;237
9.8;Chapter 24. Chemical deglycosylation of Horseradish peroxidase and surglycosylation using a new glycosylating reagent: effects on catalytic activity and stability;238
9.8.1;1. INTRODUCTION;238
9.8.2;2. RESULTS AND DISCUSSION;239
9.8.3;3. REFERENCES;244
9.9;Chapter 25. Engineering stability and specificity of the Lactococcus lactis SK11 proteinase;246
9.9.1;1. INTRODUCTION;246
9.9.2;2. MODELLING OF THE CATALYTIC DOMAIN;247
9.9.3;3. ENGINEERING OF THE SUBSTRATE BINDING REGION;248
9.9.4;4. ENGINEERING OF INSERTS IN SURFACE LOOPS;251
9.9.5;5. DISCUSSION;252
9.9.6;6. ACKNOWLEDGMENTS;253
9.9.7;7. REFERENCES;253
9.10;Chapter 26. Comparative studies on the thermophilicity and stability of 5'-methylthioadenosine phosphorylase from various sources;254
9.10.1;INTRODUCTION;254
9.10.2;SOURCES OF MTA PHOSPHORYLASE;255
9.10.3;THERMOPHILICITY AND THERMOSTABILITY;255
9.10.4;STABILITY AND EFFECT OF REDUCING AGENTS;257
9.10.5;MOLECULAR WEIGHT AND PROTEIN STRUCTURE;258
9.10.6;SUSCEPTIBILITY TO PROTEASES;258
9.10.7;CONCLUSIONS;260
9.10.8;ACKNOWLEDGMENTS;261
9.10.9;REFERENCES;261
9.11;CHAPTER 27. A PROCESS FOR STABILIZATION OF GLYCOPROTEINS;262
9.11.1;INTRODUCTION;262
9.11.2;MATERIALS AND METHODS;263
9.11.3;RESULTS;264
9.11.4;CONCLUSION;267
9.11.5;REFERENCES;268
9.12;CHAPTER 28. PRESSURE - INDUCED STRUCTURAL MODIFICATIONS OF BUTYRYLCHOLINESTERASE;270
9.12.1;1. INTRODUCTION;270
9.12.2;2. MATERIALS AND METHODS;271
9.12.3;3. RESULTS AND DISCUSSION;272
9.12.4;4.ACKNOWLEDGMENTS;274
9.12.5;5. REFERENCES;275
9.13;CHAPTER 29. CHANGING THE THERMOSTABILITY OF BACILLUS LICHENIFORMIS a-AMYLASE;276
9.13.1;1. INTRODUCTION;276
9.13.2;2. MATERIALS AND METHODS;277
9.13.3;3. RESULTS AND DISCUSSION;278
9.13.4;4. CONCLUSIONS;282
9.13.5;5. REFERENCES;282
9.14;Chapter 30. Stabilization of Lipases for Hydrolysis Reactions on Industrial Scale;284
9.14.1;1. INTRODUCTION;284
9.14.2;2. PRINCIPLE;285
9.14.3;3. REACTOR EXPERIMENTS;286
9.14.4;4. RESULTS and DISCUSSION;286
9.14.5;5. CONCLUSIONS;288
9.14.6;6. REFERENCES;288
9.15;Chapter 31. Cold denaturation of proteins as investigated by subzero transverse temperature gradient gel electrophoresis;290
9.15.1;1. INTRODUCTION;290
9.15.2;2. MATERIALS AND METHODS;291
9.15.3;3. RESULTS;293
9.15.4;4. DISCUSSION;295
9.15.5;5. CONCLUSION;296
9.15.6;6. ACKNOWLEDGEMENT;296
9.15.7;7. REFERENCES;297
9.16;Chapter 32. Genetic algorithms as a new tool to study protein stability;298
9.16.1;1. INTRODUCTION;298
9.16.2;2. MATERIALS AND METHODS;299
9.16.3;3. RESULTS;300
9.16.4;4. DISCUSSION;305
9.16.5;5. REFERENCES;305
9.17;Chapter 33. Modeling three-dimensional structure and electrostatics of alkali-stable cyclomaltodextrin glucanotransferase;306
9.17.1;1. INTRODUCTION;306
9.17.2;2. METHODS;307
9.17.3;3. RESULTS AND DISCUSSION;309
9.17.4;4. REFERENCES;313
9.18;Chapter 34. The effect of metal ion binding on protein stability;314
9.18.1;1. INTRODUCTION;314
9.18.2;2. THEORETICAL FRAMEWORK;315
9.18.3;3. THERMODYNAMIC ANALYSIS OF METAL BINDING TO a-LACTALBUMIN;318
9.18.4;4. THE NET EFFECT OF METAL ION BINDING ON PROTEIN STABILITY;321
9.18.5;5. CONCLUSIONS;322
9.18.6;Acknowledgements;322
9.18.7;References;322
9.19;Chapter 35. The number of cooperative regions (energetical domains) in a pepsin molecule depends on the pH of the medium;324
9.19.1;1. INTRODUCTION;324
9.19.2;2. MATERIALS AND METHODS;325
9.19.3;3. RESULTS AND DISCUSSION;325
9.19.4;4. CONCLUSIONS;329
9.19.5;REFERENCES;329
9.20;CHAPTER 36. STABILIZATION OF SOLUBLE PROTEINS BY INTRAMOLECULAR CROSSLINKING WITH POLYFUNCTIONAL MACROMOLECULES. POLY-(GLUTARALDEHYDE-LIKE) STRUCTURE;330
9.20.1;INTRODUCTION;330
9.20.2;MATERIALS AND METHODS;333
9.20.3;DEXTRAN OXIDATION;333
9.20.4;ENZYME ACTIVITY ASSAYS;333
9.20.5;ENZYME MODIFICATION;333
9.20.6;POLYACRYLAMIDE GEL ELECTROPHORESIS IN PRESENCE OF SDS (PAGE-SDS);333
9.20.7;ENZYME STABILITY ASSAYS;334
9.20.8;DEVELOPING OF YELLOW COLOR BY DEXTRAN AT ALKALINE pHs;334
9.20.9;RESULTS;334
9.20.10;POLY-(GLUTARALDEHYDE-LIKE) MODIFICATION EFFECT ON INDUSTRIAL ENZYME PROPERTIES;335
9.20.11;LINEAR POLY-(GLUTARALDEHYDE-LIKE) STRUCTURE PRESERVATION;336
9.20.12;DISSCUSION;337
9.20.13;REFERENCES;337
9.21;Chapter 37. Development of A Method for the Stabilization and Formulation of Xylanase from Trichoderma Using Experimental Design;338
9.21.1;Introduction;338
9.21.2;Materials and Methods;339
9.21.3;Results;339
9.21.4;Discussion;343
9.22;Chapter 38. New technique for monitoring interfacial inactivation of enzymes by organic solvents;344
9.22.1;INTRODUCTION;344
9.22.2;APPARATUS AND METHODOLOGY;345
9.22.3;TYPES OF BEHAVIOUR OBSERVED;346
9.22.4;EFFECTS OF DIFFERENT SOLVENTS;349
9.22.5;ACKNOWLEDGEMENTS;351
9.22.6;REFERENCES;351
9.23;CHAPTER 39. THE STABILISATION of ANALYTICAL ENZYMES using POLYELECTROLYTES and SUGAR DERIVATIVES;352
9.23.1;SUMMARY;352
9.23.2;INTRODUCTION;352
9.23.3;MATERIALS and METHODS;352
9.23.4;RESULTS and DISCUSSION;354
9.23.5;CONCLUSIONS;360
9.23.6;REFERENCES;360
9.24;Chapter 40. Pressure effect on the stability of Lipoxygenase: FTIR studies with the diamond anvil cell;362
9.24.1;1. INTRODUCTION;362
9.24.2;2. MATERIALS AND METHODS;364
9.24.3;3. RESULTS AND DISCUSSION;365
9.24.4;4. REFERENCES;367
9.25;Chapter 41. Stability of Ca2+-binding mutants of human lysozyme;368
9.25.1;1. INTRODUCTION;368
9.25.2;2. MATERIALS AND METHODS;369
9.25.3;3. RESULTS;369
9.25.4;ACKNOWLEDGEMENT;375
9.25.5;REFERENCES;375
9.26;Chapter 42. Storage stability of enzymes in dry apolar solvent;376
9.26.1;1. INTRODUCTION;376
9.26.2;2. Materials and Methods;376
9.26.3;3. Results and Discussion;378
9.26.4;4. Conclusion;381
9.26.5;5. Acknowledgements;382
9.26.6;6. References;382
9.27;Chapter 43. Pressure Sensitivity of Enzymes and Their Modification Proteolytic Modification of Metallo Protease under High Pressure;384
9.27.1;1. INTRODUCTION;384
9.27.2;2. EXPERIMENTAIS;386
9.27.3;3. RESULTS AND DISCUSSIONS;386
9.27.4;4.REFERENCES;390
9.28;Chapter 44. Comparative study of thermostability and structure of close homologs - barnase and binase;392
9.28.1;1. INTRODUCTION;392
9.28.2;2. MATERIALS AND METHODS;392
9.28.3;3. RESULTS AND DISCUSSION;393
9.28.4;ACKNOWLEDGEMENT;397
9.28.5;REFERENCES;397
9.29;Chapter 45. Immobilized Concanavalin A decreases the stability atproteolysis of amineoxidases;398
9.29.1;1. INTRODUCTION;398
9.29.2;2. EXPERIMENTAL;399
9.29.3;3. RESULTS;400
9.29.4;4. DISCUSSIONS;403
9.29.5;5. REFERENCES;404
9.30;Chapter 46. Stabilisation of lipase from Candida rugosa by covalent immobilisation;406
9.30.1;1. INTRODUCTION;406
9.30.2;2. MATERIALS AND METHODS;407
9.30.3;3. RESULTS AND DISCUSSION;408
9.30.4;4. CONCLUSIONS;413
9.30.5;5. REFERENCES;413
9.31;Chapter 47. Isolation, characterization and immobilization of penicillin acylase from Escherichia coli B-130;414
9.31.1;1. INTRODUCTION;414
9.31.2;2. MATERIALS AND METHODS;415
9.31.3;3. RESULTS AND DISCUSSION;416
9.31.4;4. REFERENCES;420
9.32;Chapter 48. Thermodynamic properties of apocytochrome P450Cam;422
9.32.1;Introduction;422
9.32.2;Materials and Methods;423
9.32.3;Results;423
9.32.4;Discussion;427
9.32.5;References;428
9.33;CHAPTER 49. STABILIZATION OF YEAST D-AMINO ACID OXIDASE BY MATRIX COVALENT ATTACHMENT;430
9.33.1;1. INTRODUCTION;430
9.33.2;2. EXPERIMENTAL;431
9.33.3;3. RESULTS AND DISCUSSION;432
9.33.4;4. ACKNOWLEDGEMENTS;435
9.33.5;5. REFERENCES;435
9.34;Chapter 50. Stability of an entrapped-cell system for the .1-dehydrogenation of steroids in organic medium;436
9.34.1;1. INTRODUCTION;436
9.34.2;2. MATERIALS AND METHODS;437
9.34.3;3. RESULTS AND DISCUSSION;438
9.34.4;4. CONCLUSIONS;443
9.34.5;5. REFERENCES;443
9.35;Chapter 51. The kinetics of enzyme inactivation;444
9.35.1;INTRODUCTION;444
9.35.2;EXPERIMENTAL SYSTEMS;444
9.35.3;EXPERIMENTAL RESULTS;445
9.35.4;CONCLUSIONS;450
9.35.5;LITERATURE;450
9.36;Chapter 52. Studies on stability of S-adenosylhomocysteine hydrolase from Sulfolobus solfataricus, a thermophilic archaebacterium;452
9.36.1;INTRODUCTION;452
9.36.2;THERMOPHILICITY AND THERMOSTABILITY;453
9.36.3;ENZYME STABILITY;455
9.36.4;RESISTANCE TO PROTEOLYSIS;457
9.36.5;ACKNOWLEDGMENTS;458
9.36.6;REFERENCES;458
9.37;Chapter 53. Temperature, pH and Media Influence on Lipase Stability;460
9.37.1;1. INTRODUCTION;460
9.37.2;2. MATERIALS AND METHODS;461
9.37.3;3. RESULTS;462
9.37.4;4. CONCLUSIONS;465
9.37.5;5. REFERENCES;465
9.38;Chapter 54. Prediction of the unfolding heat capacity change of proteins;466
9.38.1;1. INTRODUCTION;466
9.38.2;2. METHODS;467
9.38.3;3. RESULTS AND DISCUSSION;467
9.38.4;4. ACKNOWLEDGEMENT;472
9.38.5;4. REFERENCES;472
9.39;Chapter 55. Inactivation of a-amylase from Bacillus amyloliquefaciens at low moisture contents;474
9.39.1;ABSTRACT;474
9.39.2;1. INTRODUCTION;474
9.39.3;2. MATERIALS AND METHODS;475
9.39.4;3. RESULTS AND DISCUSSION;477
9.39.5;4. CONCLUSION;480
9.39.6;5. REFERENCES;481
9.40;CHAPTER 56. KINETICS OF HIGH-TEMPERATURE INACTIVATION OF EXTRACELLULAR PROTEASE FROM PSEUDOMONAS FLUORESCENS 22F;482
9.40.1;1. INTRODUCTION;482
9.40.2;2. MATERIALS AND METHODS;483
9.40.3;3. RESULTS AND DISCUSSION;484
9.40.4;4. REFERENCES;487
9.41;Chapter 57. Stability of a Fusarium solani pisi recombinant cutinase in reversed micelles;488
9.41.1;1. INTRODUCTION;488
9.41.2;2. MATERIALS AND METHODS;489
9.41.3;3. RESULTS AND DISCUSSION;490
9.41.4;4. CONCLUSIONS;494
9.41.5;5. REFERENCES;494
9.42;Chapter 58. Influence of long chain alcohols and chemical modification on the microencapsulated a-chymotrypsin stability;496
9.42.1;1. INTRODUCTION;496
9.42.2;2. MATERIALS AND METHODS;497
9.42.3;3. RESULTS AND DISCUSSION;497
9.42.4;4. CONCLUSIONS;502
9.42.5;5. ACKNOWLEDGEMENTS;503
9.42.6;6. REFERENCES;503
9.43;CHAPTER 59. STABILITY OF A. ORYZAE ß-GALACTOSIDASE IN WATER MISCIBLE ORGANIC SOLVENTS;504
9.43.1;INTRODUCTION;504
9.43.2;METHODS;504
9.43.3;RESULTS AND DISCUSSION;504
9.43.4;REFERENCES;511
9.44;Chapter 60. Fixation of the unfolding region - a hypothesis of enzyme stabilization;512
9.44.1;1. INTRODUCTION;512
9.44.2;2. Biphasic thermal inactivation kinetics of immobilized enzymes;512
9.44.3;3. Influence of temperature on the thermal inactivation kinetics;514
9.44.4;4. Evidence of subpopulations by spin and fluorescence labeling;515
9.44.5;5. Model of unfolding region;517
9.44.6;6. ACKNOWLEDGEMENT;519
9.44.7;7. REFERENCES;519
9.45;CHHAPTER 61. PROBING CONFORMATIONAL TRANSITIONS IN INTERFACE a1ß2 OF HUMAN HEMOGLOBIN BY SITE-DIRECTED MUTAGENESIS;520
9.45.1;Introduction;520
9.45.2;Results;520
9.45.3;Mutagenesis and expression in E. coli;522
9.45.4;Ultraviolet differential spectra;522
9.45.5;Determination of dissociation constants for the equilibrium a2ß2 Û 2 aß using the flash photolysis method;523
9.45.6;Conclusions;523
9.45.7;Acknowledgements;524
9.45.8;References;524
9.46;Chapter 62. Chemical modification of the protein molecules improve their activity in organic solvents;526
9.46.1;1. INTRODUCTION;526
9.46.2;2. MATERIALS AND METHODS;527
9.46.3;3. RESULTS AND DISCUSSION;527
9.46.4;PREFERENCES;531
10;AUTHOR INDEX;534