Biocatalysis for Practitioners

Techniques, Reactions and Applications
1. Auflage 2021
ISBN: 978-3-527-34683-7
Verlag: WILEY-VCH

Buch, Englisch, 528 Seiten, Format (B × H): 170 mm x 244 mm, Gewicht: 1002 g

This reference book originates from the interdisciplinary research cooperation between academia and industry. In three distinct parts, latest results from basic research on stable enzymes are explained and brought into context with possible industrial applications. Downstream processing technology as well as biocatalytic and biotechnological production processes from global players display the enormous potential of biocatalysts. Application of "extreme" reaction conditions (i.e. unconventional, such as high temperature, pressure, and pH value) - biocatalysts are normally used within a well defined process window - leads to novel synthetic effects. Both novel enzyme systems and the synthetic routes in which they can be applied are made accessible to the reader. In addition, the complementary innovative process technology under unconventional conditions is highlighted by latest examples from biotech industry.

Autoren/Hrsg.

de Gonzalo, Gonzalo

Lavandera, Iván

Fachgebiete

• Naturwissenschaften Chemie Organische Chemie
• Technische Wissenschaften Verfahrenstechnik | Chemieingenieurwesen | Biotechnologie Biotechnologie
• Naturwissenschaften Chemie Physikalische Chemie Biokatalyse

Inhaltsverzeichnis

Foreword xvii

Part I Enzyme Techniques 1

1 Techniques for Enzyme Purification 3
Adrie H. Westphal and Willem J. H. van Berkel

1.1 Introduction 3

1.2 Traditional Enzyme Purification 4

1.2.1 Ion Exchange Chromatography 7

1.2.2 Gel Filtration 9

1.2.3 Bio-affinity Chromatography 11

1.2.4 Hydrophobic Interaction Chromatography 14

1.2.5 Hydroxyapatite Chromatography 15

1.3 Example of a Traditional Enzyme Purification Protocol 17

1.4 Purification of Recombinant Enzymes 18

1.4.1 Immobilized Metal Affinity Chromatography 18

1.4.2 Affinity Chromatography with Protein Tags 20

1.5 Column Materials 22

1.6 Conclusions 24  References 25

2 Enzyme Modification 33
Antonino Biundo, Patricia Saénz-Méndez, and Tamas Görbe

2.1 Introduction 33

2.2 Practical Approach: Experimental Information, Analytical Methods, Tips and Tricks, and Examples 34

2.2.1 Directed Evolution 34

2.2.1.1 (Ultra)High-Throughput Screening and Selection 35

2.2.1.2 Applications of Directed Evolution Methodology 36

2.2.2 Semi-rational Design 37

2.2.2.1 Applications of Semi-rational Design Methodology 38

2.2.3 De Novo Enzyme Design 39

2.2.3.1 Applications of De Novo Enzyme Design Methodology 40

2.2.4 Rational Enzyme Design 40

2.2.4.1 Applications of Rational Design Methodology 41

2.3 Expectations and Perspectives 49

2.4 Concluding Remarks 50

References 51

3 Immobilization Techniques for the Preparation of Supported Biocatalysts: Making Better Biocatalysts Through Protein Immobilization 63
Javier Rocha-Martín, Lorena Betancor, and Fernando López-Gallego

3.1 Introduction 63

3.2 General Aspects to Optimize Enzyme Immobilization Protocols 64

3.2.1 Carrier Nature 64

3.2.2 Immobilization Chemistry 64

3.2.3 Protein Orientation 64

3.2.4 Multivalence of the Protein Attachment 65

3.2.5 Chemical and Geometrical Congruence 65

3.2.6 Enzyme Spatial Organization 65

3.3 Type of Carriers for Immobilized Proteins 66

3.3.1 Types of Materials 66

3.3.1.1 Organic Materials 66

3.3.1.2 Inorganic Materials 66

3.3.2 Geometry 67

3.3.2.1 Beads 67

3.3.2.2 Monoliths 67

3.3.2.3 Membranes 67

3.3.3 Dimensions 67

3.3.4 Commercially Available Porous Carriers for Enzyme Immobilization 68

3.4 Immobilization Methods and Manners 68

3.5 Evaluation of the Enzyme Immobilization Process 70

3.5.1 Considerations Before Immobilization 71

3.5.1.1 Preparation of the Enzymatic Solution to Be Immobilized 71

3.5.1.2 Stability of the Soluble Enzyme Under Immobilization Conditions 71

3.5.2 Parameters Required to Define an Immobilization Process 71

3.5.2.1 Immobilization Yield 72

3.5.2.2 Expressed Activity or Apparent Activity 72

3.5.2.3 Specific Activity of the Immobilized Biocatalyst 73

3.6 Applied Examples of Immobilized Enzymes 73

3.6.1 Characterization of the Immobilized Biocatalyst 74

3.6.1.1 Determination of the Catalytic Activity of the Final Immobilized Biocatalyst and Maximum Protein Loading Capacity 74

3.6.1.2 Apparent Kinetic Parameters of the Immobilized Enzyme 76

3.6.1.3 Biocatalyst Stability 77

3.6.1.3.1 The Half-life Time of Biocatalysts 78

3.7 Challenges and Opportunities in Enzyme Immobilization 79

3.8 Conclusions 81

List of Abbreviations 82

References 82

4 Compartmentalization in Biocatalysis 89
Robert Kourist and Javier González-Sabín

4.1 Introduction 89

4.2 Cell as a Compartment 93

4.3 Compartmentalization Using Protein Assemblies 95

4.4 Compartmentalization Using Emulsion and Micellar Systems 96

4.5 Compartmentalization Using Encapsulation 100

4.6 Compartmentalization Using Tea Bags and Thimbles 103

4.7 Separation of Reaction Steps Using Continuous Flow 105

4.8 Conclusions and Prospects 107

References 108

Part II Enzymes Handling and Applications 113

5 Promiscuous Activity of Hydrolases 115
Erika V. M. Orozco and André L. M. Porto

5.1 Introduction 115

5.2 Catalytic Promiscuity 116

5.3 Hydrolases 117

5.3.1 Applications of Hydrolases to Organic Synthesis 118

5.3.2 Lipases and Their Hydrolysis Mechanism 122

5.3.3 Catalytic Promiscuity of Hydrolases 122

5.3.4 Promiscuous Aldol Reaction Catalyzed by Hydrolases 130

5.3.5 Aldol Reaction Between 4-Cyanobenzaldehyde and Cyclohexanone Catalyzed by Porcine Pancreatic Lipase (PPL-II) and Rhizopus niveus Lipase (RNL) 135

5.4 Conclusions 136

References 137

6 Enzymes Applied to the Synthesis of Amines 143
Francesco G. Mutti and Tanja Knaus

6.1 Introduction 143

6.2 Hydrolases 145

6.2.1 Practical Approaches with Hydrolases 145

6.2.1.1 Kinetic Resolution 145

6.2.1.2 Dynamic Kinetic Resolution 146

6.2.2 Practical Examples with Hydrolases 148

6.2.2.1 Kinetic Resolution of Racemic a-Methylbenzylamine Through the Methoxyacetylation Catalyzed by a Lipase 148

6.2.2.2 Dynamic Kinetic Resolution for the Synthesis of Norsertraline 149

6.3 Amine Oxidases 149

6.3.1 Practical Approaches with Amine Oxidases 150

6.3.1.1 Kinetic Resolution and Deracemization 150

6.3.2 Practical Examples with Amine Oxidases 151

6.3.2.1 One-pot, One-enzyme Oxidative Pictet–Spengler Approach Combined with Deracemization 151

6.3.2.2 Desymmetrization of meso-compounds 152

6.4 Transaminases (or Aminotransferases) 152

6.4.1 Practical Approaches with Transaminases 153

6.4.2 Practical Examples with Transaminases 153

6.4.2.1 Kinetic Resolution and Deracemization 153

6.4.2.2 Asymmetric Synthesis from Prochiral Ketone 155

6.5 Am

Über Autor(innen)

Gonzalo de Gonzalo is Associate Professor at the Departamento de Química at the University of Seville, Spain. He obtained his Degree and his Ph. D. in chemistry at the University of Oviedo (Spain). He spent his postdoctoral stage at Consiglio Nazionale delle Ricerche (Milan, Italy), moving back to University of Oviedo with a Juan de la Cierva Fellowship. In 2010, he spent one year at the University of Groningen working tin the research of novel oxidative enzymes. He worked two years for the pharmaceutical company Antibióticos S.A.U. (León, Spain), moving to Seville in 2014. His research is focused on asymmetric synthesis by using different approaches, including biocatalytic and organocatalytic procedures, as well as the development of concurrent chemo- and biocatalytic reactions. He has published over 70 scientific publications and has recently been coeditor of the book "Biocatalysis: an Industrial Perspective".

Iván Lavandera completed his Ph. D. studies in Chemistry in 2003 with Prof. Vicente Gotor and Prof. Miguel Ferrero. He continued in Prof. Gotor's group as a researcher until 2005, and then he moved to the University of Graz as a postdoctoral researcher under the supervision of Prof. Wolfgang Kroutil. He returned to Oviedo in 2008, where he became first as a Clarín and then as a Ramón y Cajal post-doctoral researcher. Since 2015, he is Associate Professor at the Organic and Inorganic Department at the University of Oviedo, where he got the habilitation in 2017. He has been co-author of two patents and more than 90 publications. His research interests are focused on Biocatalysis, especially the use of oxidoreductases and transferases to achieve green processes and to develop new synthetic routes combining bio- and chemocatalysis in a concurrent manner.