About Book

Enzymes are very important for metabolic processes and human survival. They are macromolecular biological catalysts and are responsible for the acceleration of chemical reactions. The biological processes that occur within all living organisms are chemical reactions, and most are regulated by enzymes. Majority of functional enzymes are proteins, like trypsin, fumarase or papain. A rapid expansion of the knowledge base in the field of enzyme biotechnology has occurred over the past few years. Much of this expansion has been driven by the bio-discovery of many new enzymes from a wide range of environments, some extreme in nature, followed by subsequent protein (enzyme) engineering. These enzymes have found a wide range of applications, ranging from bioremediation, bio-monitoring, biosensor development, bioconversion to biofuels and other biotechnologically important value-added products. The book aims to shed light on some of the unexplored aspects of enzymes. The topics included in this book on enzymes are of utmost significance and bound to provide incredible insights to readers. It deals with antioxidant enzymes, biodegradation, oxidative enzymes, actinobacteria, catalyzed biotransformation, sterol biosynthetic pathway, enzyme-assisted extraction and luciferase enzymes. It unfolds the innovative aspects of this field which will be crucial for the holistic understanding of the subject matter. While understanding the long-term perspectives of the topics, the book makes an effort in highlighting their impact as a modern tool for the growth of the discipline. It will serve as a valuable source of reference for those interested in this field.

Preface............................................................................................................v

Chapter 1 Biocatalysis and Strategies for Enzyme Improvement......................1

Abstract..........................................................................................................1

1. Introduction............................................................................................2

2. Improving biocatalysis in organic soLvents by additive

approach.....................................................................................................3

3. Chemical modifications to stabilize enzymes....................................6

4. Enzyme immobilization.......................................................................7

5. Enzyme engineering...........................................................................14

6. Conclusions..........................................................................................19

Conflict of interest..............................................................................19

Notes/thanks/other declarations.......................................................19

Chapter 2 Biotechnological Advances in Luciferase Enzymes..........................31

Abstract.................................................................................................31

1. Introduction..........................................................................................31

2. Firefly and click beetle luciferases...................................................34

3. Renilla luciferase.................................................................................38

4. Gaussia luciferase.................................................................................41

5. Oplophorus luciferase...........................................................................43

6. Bacterial luciferase..............................................................................45

7. Fungal luciferase.................................................................................49

8. Outlook for future developments....................................................51

9. Conclusion............................................................................................52

Acknowledgements.............................................................................52

Conflict of interest..............................................................................52

Chapter 3 Biopesticide of Neem Obtained by Enzyme-Assisted Extraction:

An Alternative to Improve the Pest Control......................................62

Abstract.................................................................................................62

1. Introduction..........................................................................................63

2. Assisted enzymatic extraction of neem..........................................66

3. Results and discussion.......................................................................68

4. Conclusions..........................................................................................75

Acknowledgements.............................................................................76

Conflict of interest..............................................................................76

Chapter 4 3-Hydroxy-3-Methylglutaryl-CoA Reductase (HMGR) Enzyme

of the Sterol Biosynthetic Pathway: A Potential Target against

Visceral Leishmaniasis ...........................................................................81

Abstract.................................................................................................81

1. Introduction..........................................................................................82

2. HMGR inhibition profiles..................................................................85

3. Natural products as inhibitors of HMGR......................................87

Chapter 5 Kinetic Modelling of Enzyme Catalyzed Biotransformation

Involving Activations and Inhibitions................................................92

Abstract.................................................................................................92

1. Introduction..........................................................................................92

2. Study of enzyme mechanism and kinetics....................................97

3. Enzyme inhibition.............................................................................117

4. Analytical aspects of enzyme inhibition......................................128

5. Design of inhibitors and activators...............................................135

6. Enzyme activators.............................................................................139

7. New developments in enzyme inhibition....................................140

8. Conclusion..........................................................................................142

Abbreviations.....................................................................................143

Chapter 6 Actinobacteria — A Biofactory of Novel Enzymes........................154

Abstract...............................................................................................154

1. Introduction........................................................................................155

2. Significance of actinobacterial enzymes.......................................156

3. Types of actinobacterial enzymes..................................................158

4. Conclusion..........................................................................................170

Chapter 7 Biotechnological Potential of Oxidative Enzymes from

Actinobacteria..........................................................................................181

Abstract...............................................................................................181

1. Introduction........................................................................................181

2. Oxidative enzymes produced by actinobacterial strains..........183

3. Industrial relevance of actinobacterial oxidases: how to access

them and improve their functionality..........................................193

Chapter 8 Biodegradation of Melanoidin from Distillery Effluent: Role of

Microbes and Their Potential Enzymes............................................215

1. Introduction........................................................................................215

2. Nature of melanoidins.....................................................................217

3. Degradation of melanoidin polymer.............................................219

4. Biological methods for the treatment of distillery effluent......221

5. Role of enzymes in effluent decolorization.................................240

Chapter 9 Plant Antioxidative Enzymes – Case Study: In Vitro Organogenesis

of Saffron (Crocus sativus L.)..............................................................257

1. Introduction........................................................................................257

2. Plant media.........................................................................................260

3. Protein extraction and protein assay............................................261

4. Enzyme activity.................................................................................262

5. Results.................................................................................................263

6. Isozyme banding patterns...............................................................266

7. Conclusion..........................................................................................267

Chapter 10 Antioxidant Enzymes and Human Health.......................................271

1. Introduction........................................................................................271

2. Free radicals and their scavengers................................................272

3. Reactive Oxygen Species.................................................................272

4. Antioxidant protection system.......................................................275

5. Antioxidant enzymes in health......................................................277

6. Clinical applications of antioxidant enzymes..............................283

7. Other antioxidants............................................................................285

8. Conclusion..........................................................................................286

Citation......................................................................................................291

Index...........................................................................................................293

About Author

Cortez Rosales holds Ph.D. in Biomedical and Life Science from Public Science University, Catalonia. She serves as a Post-Doctoral Fellow and Research Engineer at Centre for Scientific Research, Rennes. Her primary areas of scholarly and research interests lie in the fields of enzymology and metabolic processes. Rosales has authored and edited numerous articles, journal papers and book chapters, which have been published worldwide. She has attende d numerous scientific conferences and symposiums and has presented 8 papers on enzymology.