Full Description
The whole range of biocatalysis, from a firm grounding in theoretical concepts to in-depth coverage of practical applications and future perspectives.
The book not only covers reactions, products and processes with and from biological catalysts, but also the process of designing and improving such biocatalysts.
One unique feature is that the fields of chemistry, biology and bioengineering receive equal attention, thus addressing practitioners and students from all three areas.
Contents
Preface v
Acknowledgments vii
1 Introduction to Biocatalysis 1
1.1 Overview:The Status of Biocatalysis at the Turn of the 21st Century 2
1.2 Characteristics of Biocatalysis as a Technology 6
1.3 Current Penetration of Biocatalysis 11
1.4 The Breadth of Biocatalysis 14
2 Characterization of a (Bio-)catalyst 19
2.1 Characterization of Enzyme Catalysis 20
2.2 Sources and Reasons for the Activity of Enzymes as Catalysts 23
2.3 Performance Criteria for Catalysts, Processes, and Process Routes 30
3 Isolation and Preparation of Microorganisms 43
3.1 Introduction 44
3.2 Screening of New Enzyme Activities 46
3.3 Strain Development 48
3.4 Extremophiles 52
3.5 Rapid Screening of Biocatalysts 56
4 Molecular Biology Tools for Biocatalysis 61
4.1 Molecular Biology Basics: DNA versus Protein Level 62
4.2 DNA Isolation and Purification 65
4.3 Gene Isolation, Detection, and Verification 67
4.4 Cloning Techniques 77
4.5 (Over)expression of an Enzyme Function in a Host 81
5 Enzyme Reaction Engineering 91
5.1 Kinetic Modeling: Rationale and Purpose 92
5.2 The Ideal World: Ideal Kinetics and Ideal Reactors 94
5.3 Enzymes with Unfavorable Binding: Inhibition 97
5.4 Reactor Engineering 105
5.5 Enzyme Reactions with Incomplete Mass Transfer: Influence of Immobilization 113
5.6 Enzymes with Incomplete Stability: Deactivation Kinetics 119
5.7 Enzymes with Incomplete Selectivity: E-Value and its Optimization 126
6 Applications of Enzymes as Bulk Actives: Detergents, Textiles, Pulp and Paper, Animal Feed 135
6.1 Application of Enzymes in Laundry Detergents 136
6.2 Enzymes in the Textile Industry: Stone-washed Denims, Shiny Cotton Surfaces 140
6.3 Enzymes in the Pulp and Paper Industry: Bleaching of Pulp with Xylanases or Laccases 145
6.4 Phytase for Animal Feed: Utilization of Phosphorus 152
7 Application of Enzymes as Catalysts: Basic Chemicals, Fine Chemicals, Food, Crop Protection, Bulk Pharmaceuticals 159
7.1 Enzymes as Catalysts in Processes towards Basic Chemicals 160
7.2 Enzymes as Catalysts in the Fine Chemicals Industry 170
7.3 Enzymes as Catalysts in the Food Industry 187
7.4 Enzymes as Catalysts towards Crop Protection Chemicals 195
7.5 Enzymes for Large-Scale Pharma Intermediates 197
8 Biotechnological Processing Steps for Enzyme Manufacture 209
8.1 Introduction to Protein Isolation and Purification 210
8.2 Basics of Fermentation 212
8.3 Fermentation and its Main Challenge: Transfer of Oxygen 218
8.4 Downstream Processing: Crude Purification of Proteins 223
8.5 Downstream Processing: Concentration and Purification of Proteins 231
8.6 Examples of Biocatalyst Purification 237
9 Methods for the Investigation of Proteins 243
9.1 Relevance of Enzyme Mechanism 244
9.2 Experimental Methods for the Investigation of an Enzyme Mechanism 245
9.3 Methods of Enzyme Determination 253
9.4 Enzymatic Mechanisms: General Acid-Base Catalysis 258
9.5 Nucleophilic Catalysis 261
9.6 Electrophilic catalysis 269
10 Protein Engineering 281
10.1 Introduction: Elements of Protein Engineering 282
10.2 Methods of Protein Engineering 283
10.3 Glucose (Xylose) Isomerase (GI) and Glycoamylase: Enhancement of Thermostability 289
10.4 Enhancement of Stability of Proteases against Oxidation and Thermal Deactivation 293
10.5 Creating New Enzymes with Protein Engineering 295
10.6 Dehydrogenases, Changing Cofactor Specificity 298
10.7 Oxygenases 300
10.8 Change of Enantioselectivity with Site-Specific Mutagenesis 302
10.9 Techniques Bridging Different Protein Engineering Techniques 303
11 Applications of Recombinant DNA Technology: Directed Evolution 309
11.1 Background of Evolvability of Proteins 310
11.2 Process steps in Directed Evolution: Creating Diversity and Checking for Hits 314
11.3 Experimental Protocols for Directed Evolution 319
11.4 Successful Examples of the Application of Directed Evolution 325
11.5 Comparison of Directed Evolution Techniques 331
12 Biocatalysis in Non-conventional Media 339
12.1 Enzymes in Organic Solvents 340
12.2 Evidence for the Perceived Advantages of Biocatalysts in Organic Media 341
12.3 State of Knowledge of Functioning of Enzymes in Solvents 344
12.4 Optimal Handling of Enzymes in Organic Solvents 351
12.5 Novel Reaction Media for Biocatalytic Transformations 355
12.6 Solvent as a Parameter for Reaction Optimization ("Medium Engineering") 366
13 Pharmaceutical Applications of Biocatalysis 373
13.1 Enzyme Inhibition for the Fight against Disease 374
13.2 Enzyme Cascades and Biology of Diseases 380
13.3 Pharmaceutical Applications of Biocatalysis 393
13.4 Applications of Specific Biocatalytic Reactions in Pharma 402
14 Bioinformatics 413
14.1 Starting Point: from Consequence (Function) to Sequence 414
14.2 Bioinformatics: What is it, Why do we Need it, and Why Now? (NCBI Homepage) 415
14.3 Tools of Bioinformatics: Databases, Alignments, Structural Mapping 418
14.4 Applied Bioinformatics Tools, with Examples 422
14.5 Bioinformatics for Structural Information on Enzymes 429
14.6 Conclusion and Outlook 431
15 Systems Biology for Biocatalysis 433
15.1 Introduction to Systems Biology 434
15.2 Genomics, Proteomics, and other -omics 435
15.3 Technologies for Systems Biology 438
15.4 Metabolic Engineering 449
16 Evolution of Biocatalytic Function 457
16.1 Introduction 458
16.2 Search Characteristics for Relatedness in Proteins 461
16.3 Evolution of New Function in Nature 466
16.4 α/β-Barrel Proteins as a Model for the Investigation of Evolution 474
17 Stability of Proteins 487
17.1 Summary: Protein Folding, First-Order Decay, Arrhenius Law 488
17.2 Two-State Model: Thermodynamic Stability of Proteins (Unfolding) 491
17.3 Three-State Model: Lumry-Eyring Equation 493
17.4 Four-State Model: Protein Aggregation 496
17.5 Causes of Instability of Proteins: ∆G < 0, γ(t), A 501
17.6 Biotechnological Relevance of Protein Folding: Inclusion Bodies 505
17.7 Summary: Stabilization of Proteins 506
18 Artificial Enzymes 511
18.1 Catalytic Antibodies 512
18.2 Other Proteinaceous Catalysts: Ribozymes and Enzyme Mimics 521
18.3 Design of Novel Enzyme Activity: Enzyme Models (Synzymes) 523
18.4 Heterogenized/Immobilized Chiral Chemical Catalysts 526
18.5 Tandem Enzyme Organometallic Catalysts 532
19 Design of Biocatalytic Processes 539
19.1 Design of Enzyme Processes: High-Fructose Corn Syrup (HFCS) 540
19.2 Processing of Fine Chemicals or Pharmaceutical Intermediates in an Enzyme Membrane Reactor 549
19.3 Production of Enantiomerically Pure Hydrophobic Alcohols: Comparison of Different Process Routes and Reactor Configurations 556
20 Comparison of Biological and Chemical Catalysts for Novel Processes 569
20.1 Criteria for the Judgment of (Bio-)catalytic Processes 570
20.2 Position of Biocatalysis in Comparison to Chemical Catalysts for Novel Processes 575
20.3 Pathway Engineering through Metabolic Engineering 586
Index 593
