High Value Fermentation Products Volume 1

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High Value Fermentation Products Volume 1

Saurabh Saran, Vikash Babu and Asha Chuabey

Human Health

Edited by

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v

Contents

Foreword xviiAbout the Editors xixList of Contributors xxiPreface xxvAcknowledgement xxvii

1 Introduction, Scope and Significance of Fermentation Technology 1Saurabh Saran, Alok Malaviya and Asha Chaubey 1.1 Introduction 1 1.2 Background of Fermentation Technology 2 1.3 Market of Fermentation Products 3 1.4 Types of Fermentation 4

1.4.1 Solid State Fermentation (SSF) 41.4.2 Submerged Fermentation (SmF) 71.4.3 Solid State (SSF) vs. Submerged (SmF) Fermentation 9

1.5 Classification of Fermentation 9 1.6 Design and Parts of Fermentors 10 1.7 Types of Fermentor 15

1.7.1 Stirred Tank Fermentor 151.7.2 Airlift Fermentor 161.7.3 Bubble Column Fermentor 171.7.4 Fluidized Bed Fermentor 181.7.5 Packed Bed Fermentor 191.7.6 Photo Bioreactor 19

1.8 Industrial Applications of Fermentation Technology 21 1.9 Scope and Global Market of Fermentation Technology 221.10 Conclusions 23References 24

2 Extraction of Bioactive Molecules through Fermentation and Enzymatic Assisted Technologies 27Ramón Larios-Cruz, Liliana Londoño-Hernández, Ricardo Gómez-García, Ivanoe García, Leonardo Sepulveda, Raúl Rodríguez-Herrera and Cristóbal N. Aguilar2.1 Introduction 272.2 Definition of Bioactives Compounds 29

2.2.1 Polyphenols and Polypeptides 29

vi Contents

2.2.2 Importance and Applications of Bioactive Compounds 292.2.3 Bioactive Peptides 31

2.3 Traditional Processes for Obtaining Bioactive Compounds 332.3.1 Soxhlet Extraction 332.3.2 Liquid-Liquid and Solid-Liquid Extraction 342.3.3 Maceration Extraction 35

2.4 Fermentation and Enzymatic Technologies for Obtaining Bioactive Compounds 352.4.1 Soft Chemistry in Bioactive Compounds 352.4.2 Biotransformation of Bioactive Compounds 362.4.3 Enzymatic and Fermentation Technologies 39

2.4.3.1 Enzymatic Technology 412.4.3.2 Fermentation Technology 43

2.5 Use of Agroindustrial Waste in the Fermentation Process 452.5.1 Cereal Wastes 462.5.2 Fruit and Plant Waste 46

2.6 General Parameters in the Optimization of Fermentation Processes 492.6.1 Response Surface Methodology 492.6.2 First-Order Model 492.6.3 Second-Order Model 49

2.7 Final Comments 52Acknowledgements 52References 52

3 Antibiotics Against Gram Positive Bacteria 61Rahul Vikram Singh, Hitesh Sharma, Anshela Koul and Vikash Babu3.1 Introduction 613.2 Target of Antibiotics Against Gram Positive Bacteria 64

3.2.1 Cell Wall Synthesis Inhibition 653.2.1.1 Penicillin 663.2.1.2 Cephalosporins 673.2.1.3 Vancomycin 693.2.1.4 Teicoplanin 693.2.1.5 Oritavancin 69

3.2.2 Protein Synthesis Inhibition 703.2.2.1 Erythromycin 703.2.2.2 Clarithromycin 713.2.2.3 Tetracyclines 71

3.2.3 DNA Synthesis Inhibition 723.3 Antibiotics Production Processes 723.4 Conclusion 75References 76

4 Antibiotic Against Gram-Negative Bacteria 79Maryam Faiyaz, Shikha Gupta and Divya Gupta4.1 Introduction 79

Contents vii

4.2 Gram-Negative Bacteria and Antibiotics 804.2.1 β-Lactam Drugs 81

4.2.1.1 Cephalosporins 824.2.1.2 Penicillin 824.2.1.3 Carbapenems 824.2.1.4 Monobactam 82

4.2.2 Macrolide 824.2.3 Aminoglycosides 844.2.4 Fluoroquinolones 84

4.3 Production of Antibiotics 854.3.1 Strain Development 85

4.3.1.1 Mutagenesis 864.3.1.2 Protoplast Fusion 864.3.1.3 Recombinant DNA 87

4.3.2 Media Formulation and Optimization 884.3.3 Fermentation 904.3.4 Downstream Processing and Purification 924.3.5 Quality Control 95

4.4 Conclusion 95References 96

5 Role of Antifungal Drugs in Combating Invasive Fungal Diseases 103Kakoli Dutt5.1 Introduction 1035.2 Antifungal Agents 105

5.2.1 Azoles 1145.2.2 Polyenes 1155.2.3 Allylamine/Thiocarbonates 1165.2.4 Other Antifungal Agents 117

5.3 Targets of Antifungal Agents 1205.3.1 Cell Wall Biosynthesis Inhibitors 1205.3.2 Sphingolipid Synthesis Inhibitors 1235.3.3 Ergosterol Synthesis Inhibitors 1255.3.4 Protein Synthesis Inhibitors 1265.3.5 Novel Targets 128

5.4 Development of Resistance towards Antifungal Agents 1305.4.1 Minimum Inhibitory Concentration 1305.4.2 Antifungal-Drug-Resistance Mechanisms 131

5.5 Market and Drug Development 1345.6 Conclusions 136Acknowledgement 137References 137

6 Current Update on Rapamycin Production and Its Potential Clinical Implications 145Girijesh K. Patel, Ruchika Goyal1 and Syed M. Waheed6.1 Introduction 145

viii Contents

6.2 Biosynthesis of Rapamycin 1466.2.1 Microbial Strain 1476.2.2 Optimization of Carbon, Nitrogen Sources and Salts 1476.2.3 Strain Manipulation to Improve Rapamycin Production 148

6.2.3.1 Random Mutagenesis 1486.2.3.2 Chemical Mutagenesis 1506.2.3.3 Metabolic Engineering Combined with Genetic

Engineering to Improve Rapamycin Production 1506.2.3.4 Protoplasts Fusion 1516.2.3.5 Optimization of Fermentation Methods 151

6.3 Organic Synthesis of Rapamycin 1526.4 Extraction and Quantification of Rapamycin 1526.5 Physiological Factors Affecting Rapamycin Biosynthesis 153

6.5.1 Effect of Media Components 1536.5.2 Effect of pH on Rapamycin Production 1536.5.3 Effect of Physical Gravity 1546.5.4 Effect of Morphological Changes 1546.5.5 Effect of Dissolved Oxygen (DO) and Carbon Dioxide (DCO2) 154

6.6 Production of Rapamycin Analogs 154 6.7 Mechanism of Action of Rapamycin 155 6.8 Use of Rapamycin in Medicine 157

6.8.1 Anti-Fungal Agent 1576.8.2 Immunosuppression 1586.8.3 Anti-Cancer Agent 1586.8.4 Anti-Aging Agent 1586.8.5 Role in HIV Treatment 1586.8.6 Rheumatoid Arthritis 159

6.9 Side Effects of Long-term Use of Rapamycin 1596.10 Conclusions 159Acknowledgements 160References 160

7 Advances in Production of Therapeutic Monoclonal Antibodies 165Richi V Mahajan, Subhash Chand, Mahendra Pal Singh, Apurwa Kestwal and Surinder Singh7.1 Introduction 1657.2 Discovery and Clinical Development 1667.3 Structure and Classification 1677.4 Nomenclature of Monoclonal Antibodies 1687.5 Production of Monoclonal Antibodies 170

7.5.1 Hybridoma Technology 1707.5.2 Epstein-Barr Virus Technology 1727.5.3 Phage Display Technology 1727.5.4 Cell Line Based Production Techniques 173

7.5.4.1 Cell Lines 1737.5.4.2 Production Media 1757.5.4.3 Production Conditions 175

Contents ix

7.5.4.4 Production Strategies 1767.5.4.5 Production Vessel 1777.5.4.6 Microbial Production 1807.5.4.7 Plant-Based Production 182

7.5.5 Chemical Modifications of Monoclonal Antibodies 1837.5.6 Advances in Antibody Technology 183

7.5.6.1 Antibody–Drug Conjugates (ADCs) 1837.5.6.2 Antibody Fusion Proteins 1847.5.6.3 Payloads 184

7.6 Conclusions 185References 186

8 Antimicrobial Peptides from Bacterial Origin: Potential Alternative to Conventional Antibiotics 193Lipsy Chopra, Gurdeep Singh, Ramita Taggar, Akanksha Dwivedi, Jitender Nandal, Pradeep Kumar and Debendra K. Sahoo8.1 Introduction 1938.2 Classification of Bacteriocins 194

8.2.1 Bacteriocins from Gram-Negative Bacteria 1948.2.2 Bacteriocins from Gram-Positive Bacteria 194

8.3 Mode of Action 1968.3.1 Pore-Forming Bacteriocins 1968.3.2 Non-Pore-Forming Bacteriocins: Intracellular Targets 198

8.4 Applications 1988.4.1 Food Bio Preservative 1988.4.2 Food Packaging (In Packaging Films) 1988.4.3 Hurdle Technology to Enhance Food Safety 1998.4.4 Therapeutic Potential 2008.4.5 Effect of Bacteriocins on Biofilms 200

8.5 Conclusions 202Acknowledgments 202Abbreviations 202References 202

9 Non-Ribosomal Peptide Synthetases: Nature’s Indispensable Drug Factories 205Richa Sharma, Ravi S. Manhas and Asha Chaubey9.1 Introduction 205

9.1.1 Non-Ribosomal Peptides as Natural Products 2059.1.2 Non-Ribosomal Peptides as Drugs 206

9.2 NRPS Machinery 2089.3 Catalytic Domains of NRPSs 208

9.3.1 Adenylation (A) Domains 2089.3.2 Thiolation (T) or PCP Domains 2099.3.3 Condensation (C) Domains 2099.3.4 Thioesterase (Te) Domains 209

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9.4 Types of NRPS 2109.4.1 Type A (Linear NRPS) 2109.4.2 Type B (Iterative NRPS) 2109.4.3 Type C (Non-linear NRPS) 210

9.5 Working of NRPSs 2109.5.1 Priming Thiolation Domain of NRPS 2119.5.2 Substrate Recognition and Activation 2119.5.3 Peptide Bond Formation between NRP Monomers 2119.5.4 Chain Termination of NRP Synthesis 2129.5.5 NRP Tailoring 212

9.6 Sources of NRPs 2139.7 Production of Non-Ribosomal Peptides 2169.8 Future Scope 218Acknowledgements 219References 219

10 Enzymes as Therapeutic Agents in Human Disease Management 225Babbal, Adivitiya, Shilpa Mohanty and Yogender Pal Khasa10.1 Introduction 22510.2 Pancreatic Enzymes 230

10.2.1 Trypsin (EC 3.4.21.4) 23010.2.2 Pancreatic Lipase (EC 3.1.1.3) 23110.2.3 Amylases (EC 3.2.1.1) 231

10.3 Oncolytic Enzymes 23210.3.1 L-Asparaginase (EC 3.5.1.1) 23210.3.2 L-Glutaminase (EC 3.5.1.2) 23310.3.3 Arginine Deiminase (ADI) (EC 3.5.3.6) 233

10.4 Antidiabetic Enzymes 23410.4.1 Glucokinase (EC2.7.1.1)

10.5 Liver Enzymes 23510.5.1 Superoxide Dismutase (SOD) (EC 1.15.1.1) 23510.5.2 Alkaline Phosphatase (ALP) (EC 3.1.3.1) 236

10.6 Kidney Disorder 23710.6.1 Uricase (EC 1.7.3.3) 23710.6.2 Urease (EC 3.5.1.5) 238

10.7 DNA- and RNA-Based Enzymes 23810.7.1 Dornase 23910.7.2 Adenosine Deaminase 24010.7.3 Ribonuclease 240

10.8 Enzymes for the Treatment of Cardiovascular Disorders 24110.8.1 The Hemostatic System 242

10.8.1.1 Primary Hemostasis 24210.8.1.2 Secondary Hemostasis 24210.8.1.3 Tertiary Hemostasis 244

10.8.2 Enzymes of the Hemostatic System 24410.8.2.1 Clotting Enzymes 24410.8.2.2 Thrombolytic Enzymes 248

Contents xi

10.9 Lysosomal Storage Disorders 251 10.9.1 α-Galactosidase A (EC 3.2.1.22) 251 10.9.2 Glucocerebrosidase (EC 3.2.1.45) 252 10.9.3 Acid Alpha-Glucosidase (GAA) (EC 3.2.1.20) 253 10.9.4 α-L-iduronidase (Laronidase) (EC 3.2.1.76) 253

10.10 Miscellaneous Enzymes 254 10.10.1 Phenylalanine Hydroxylase (EC 1.14.16.1) 254 10.10.2 Collagenase (EC 3.4.24.3) 255 10.10.3 Hyaluronidase 256 10.10.4 Bromelain 256

10.11 Conclusions 256References 257

11 Erythritol: A Sugar Substitute 265Kanti N. Mihooliya, Jitender Nandal, Himanshu Verma and Debendra K. Sahoo 11.1 Introduction 265

11.1.1 Background of Erythritol 265 11.1.2 History of Erythritol 268 11.1.3 Occurrence of Erythritol 268 11.1.4 General Characteristics 268

11.2 Chemical and Physical Properties of Erythritol 271 11.3 Estimation of Erythritol 271

11.3.1 Thin Layer Chromatography (TLC) 273 11.3.2 Colorimetric Assay for Detection of Polyols 273 11.3.3 High-Performance Liquid Chromatography (HPLC) 273 11.3.4 Capillary Electrophoresis (CE) 273

11.4 Production Methods for Erythritol 274 11.4.1 Chemical Methods for Erythritol Production 274 11.4.2 Fermentative Methods for Erythritol Production 274

11.4.2.1 Erythritol Production in Bacteria 274 11.4.2.2 Erythritol Production in Yeasts 274

11.5 Optimization of Erythritol Production 275 11.5.1 One Factor at a Time 276 11.5.2 Statistical Design Approaches 277

11.6 Toxicology of Erythritol 277 11.7 Applications of Erythritol 277

11.7.1 Confectioneries 278 11.7.2 Bakery 279 11.7.3 Pharmaceuticals 279 11.7.4 Cosmetics 279 11.7.5 Beverages 279

11.8 Precautions for Erythritol Usage 279 11.9 Global Market for Erythritol 28011.10 Conclusions 280References 281

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12 Sugar and Sugar Alcohols: Xylitol 285Bhumica Agarwal and Lalit Kumar Singh12.1 Introduction 285

12.1.1 Lignocellulosic Biomass 28612.1.2 Properties of Xylitol 28712.1.3 Occurrence and Production of Xylitol 289

12.2 Biomass Conversion Process 28912.2.1 Pretreatment Methodologies 289

12.2.1.1 Physical Pretreatment 29012.2.1.2 Physico-Chemical and Chemical Pretreatment 29112.2.1.3 Microbial Pretreatment 292

12.2.2 Enzymatic Hydrolysis 29212.2.3 Detoxification Techniques 293

12.2.3.1 Chemical Methods 29312.2.3.2 Physical Method 29512.2.3.3 Biological Method 295

12.3 Utilization of Xylose 29612.3.1 Microorganisms Utilizing Xylose 29612.3.2 Metabolism of Xylose 297

12.3.2.1 Transport of Xylose 29712.3.2.2 Metabolic Pathways 298

12.4 Process Variables 29912.4.1 Temperature and pH 29912.4.2 Substrate Concentration 30012.4.3 Aeration 301

References 303

13 Trehalose: An Anonymity Turns Into Necessity 309Manali Datta and Dignya Desai13.1 Introduction 30913.2 Trehalose Metabolism Pathways 31013.3 Physicochemical Properties and its Biological Significance 31113.4 Trehalose Production 312

13.4.1 Enzymatic Conversion to Trehalose 31213.4.2 Microbe Mediated Fermentation 314

13.4.2.1 Propionibacteria Mediated Conversion 31413.4.2.2 Micrococcus Mediated Conversion 31513.4.2.3 Brevibacterium Mediated Conversion 31513.4.2.4 Fungal Strain Mediated Fermentation 31513.4.2.5 Recombinant Strains in Trehalose Production 316

13.4.3 Purification and Detection of Trehalose in Fermentation Process 31613.5 Application of Trehalose 317

13.5.1 Role of Trehalose in Food Industries 31713.5.2 Role of Trehalose in Cosmetics and Pharmaceutics 318

13.6 Conclusions 319References 320

Contents xiii

14 Production of Yeast Derived Microsomal Human CYP450 Enzymes (Sacchrosomes) in High Yields, and Activities Superior to Commercially Available Microsomal Enzymes 323Ibidapo Stephen Williams and Bhabatosh Chaudhuri14.1 Introduction 323

14.1.1 Cytochrome P450 (CYP) Enzymes in Humans 32314.1.2 Human Cytochrome P450 Enzymes and their Role in Drug

Metabolism 32414.1.3 Requirement of Activating Proteins to Form Functional

Human CYP Enzymes 32514.1.4 Use of Yeast Biased Codons for the Syntheses of

Human Cytochrome P450 Genes 32514.1.5 Expression of Human CYP Genes in Baker’s Yeast from an

Episomal Plasmid 32514.1.6 Expression of Human CYP Genes in Baker’s Yeast from

Integrative Plasmids 32714.1.7 The ADH2 Promoter for Production of Human CYP Enzymes

in Baker’s Yeast 32714.1.8 Growth of Yeast Cells Containing Integrated Copies of CYP

Gene Expression Cassettes, Driven by the ADH2 Promoter, for Production of CYP Enzymes 328

14.2 Amounts of Microsomal CYP Enzyme Isolated from Yeast Strains Containing Chromosomally Integrated CYP Gene Expression Cassettes are far Higher than Strains Harbouring an Episomal Expression Plasmid Encoding a CYP Gene 32814.2.1 Preparation of Microsomal CYP Enzymes 32814.2.2 Measurement of the Amounts of Functional CYPs in

Microsomes Isolated from Baker’s Yeast 32914.2.3 Production of Functional Human CYP1A2 Microsomal

Enzyme from Baker’s Yeast 33014.2.4 Production of Functional Human CYP3A4 Microsomal

Enzyme from Baker’s Yeast 33014.2.5 Production of Functional Human CYP2D6 Microsomal

Enzyme from Baker’s Yeast 33114.2.6 Production of Functional Human CYP2C19 Microsomal

Enzyme from Baker’s Yeast 33214.2.7 Production of Functional Human CYP2C9 Microsomal

Enzyme from Baker’s Yeast 33314.2.8 Production of Functional Human CYP2E1 Microsomal

Enzyme from Baker’s Yeast 33314.2.9 Comments on the Production of Human CYP Enzymes

from Baker’s Yeast 33414.3 Comparison of CYP Enzyme Activity of Yeast-Derived Microsomes

(Sacchrosomes) with Commercially Available Microsomes Isolated from Insect and Bacterial Cells 336

14.3.1 Fluorescence-based Assays for Determining CYP Enzyme Activities in Isolated Microsomes 336

14.3.2 Comparison of Enzyme Activity of CYP1A2 Sacchrosomes with Commercially Available CYP1A2 Microsomes Isolated from Insect and Bacterial Cells 336

14.3.3 Comparison of Enzyme Activity of CYP2C9 Sacchrosomes with Those of Commercially Available CYP2C9 Microsomes from Insect and Bacterial Cells 337

14.3.4 Comparison of Enzyme Activity of CYP2C19 Sacchrosomes with Those of Commercially Available CYP2C19 Microsomes from Insect and Bacterial Cells 337

14.3.5 Comparison of Enzyme Activity of CYP2D6 Sacchrosomes with Those of Commercially Available CYP2D6 Microsomes from Insect and Bacterial Cells 338

14.3.6 Comparison of Enzyme Activity of CYP3A4 Sacchrosomes with Those of Commercially Available CYP3A4 Microsomes from Insect and Bacterial Cells 338

14.3.7 Comparison of Enzyme Activity of CYP2E1 Sacchrosomes with One of the Commercial CYP2E1 Microsomes Available from Insect Cells 339

14.4 IC50 Values of Known CYP Inhibitors Using Sacchrosomes, Commercial Enzymes and HLMs 339

14.5 Stabilisation of Sacchrosomes through Freeze-drying 34014.6 Conclusions 342References 345

15 Artemisinin: A Potent Antimalarial Drug 347Alok Malaviya, Karan Malhotra, Anil Agarwal and Katherine Saikia15.1 Introduction 34715.2 Biosynthesis of Artemisinin in Artemisia annua and Pathways Involved 34815.3 Yield Enhancement Strategies in A. annua 35115.4 Artemisinin Production Using Heterologous Hosts 352

15.4.1 Microbial Engineering 35215.4.2 Plant Metabolic Engineering 353

15.5 Spread of Artemisinin Resistance 35715.6 Challenges in Large-Scale Production 35815.7 Future Prospects 360References 360

16 Microbial Production of Flavonoids: Engineering Strategies for Improved Production 365Aravind Madhavan, Raveendran Sindhu, KB Arun, Ashok Pandey, Parameswaran Binod and Edgard Gnansounou16.1 Introduction 36516.2 Flavonoids 36616.3 Flavonoid Chemistry and Classes 36616.4 Health Benefits of Flavonoids 367

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Contents xv

16.5 Flavonoid Biosynthesis in Microorganism 368 16.6 Engineering of Flavonoid Biosynthesis Pathway 370 16.7 Metabolic Engineering Strategies 370 16.8 Applications of Synthetic Biology in Flavonoid Production 371 16.9 Post-modification of Flavonoids 37416.10 Purification of Flavonoids 37416.11 Conclusion 375Acknowledgements 375References 376

17 Astaxanthin: Current Advances in Metabolic Engineering of the Carotenoid 381Manmeet Ahuja, Jayesh Varavadekar, Mansi Vora, Piyush Sethia, Harikrishna Reddy and Vidhya Rangaswamy17.1 Introduction 381

17.1.1 Structure of Astaxanthin 38217.1.2 Natural vs. Synthetic Astaxanthin 38217.1.3 Uses and Market of Astaxanthin 383

17.2 Pathway of Astaxanthin 38417.2.1 Bacteria 38417.2.2 Algae 38417.2.3 Yeast 38517.2.4 Plants 386

17.3 Challenges/Current State of the Art in Fermentation/Commercial Production 386

17.4 Metabolic Engineering for Astaxanthin 38817.4.1 Bacteria 38817.4.2 Plants 39017.4.3 Synechocystis 39117.4.4 Algae 39117.4.5 Yeast 392

17.5 Future Prospects 393References 395

18 Exploitation of Fungal Endophytes as Bio-factories for Production of Functional Metabolites through Metabolic Engineering; Emphasizing on Taxol Production 401Sanjog Garyali, Puja Tandon, M. Sudhakara Reddy and Yong Wang18.1 Introduction 40118.2 Taxol: History and Clinical Impact 40318.3 Endophytes 403

18.3.1 Biodiversity of Endophytes 40518.3.2 Endophyte vs. Host Plant: the Relationship 405

18.4 The Plausibility of Horizontal Gene Transfer (HGT) Hypothesis 40718.5 Endophytes as Biological Factories of Functional Metabolites 40918.6 Taxol Producing Endophytic Fungi 41018.7 Molecular Basis of Taxol Production by Taxus Plants

(Taxol Biosynthetic Pathway) 412

xvi Contents

18.8 Metabolic Engineering for Synthesis of Taxol: Next Generation Tool 41618.8.1 Plant Cell Culture 41718.8.2 Microbial Metabolic Engineering for Synthesis of Taxol and

Its Precursors 41818.8.3 Metabolic Engineering in Heterologous Plant for Synthesis

of Taxol and Its Precursors 42018.9 Future Perspectives 421Acknowledgements 423References 423

Index 431

xvii

ForewordFrom last two decades we have witnesses unprecedented growth and development in biotechnology positioning the bioeconomy as a major indicator of advancement. Today, the global fermentation-based industry is already worth over 127 billion dollars. Based on the experience and expertise in this filed, we are trying to collect the different technologies advancement and products developed in biotechnology. This book ‘High Value Fermentation Products-Volume 1 (Human Health) is divided into various important sections related to Human Health like antibiotics, sugar & sugar alcohols,

enzymes, nutraceuticals, metabolic engineered derived products, this will help the read-ers to understand the importance of fermentation derived product for the betterment of human health. This book will also help to overcome of various bottle necks of the Industry/ scientific community and shall be useful for the betterment of the society and environment. This book will also shares an insight into the recent research, cutting edge technologies, high value products, industrial demand which bring immense interest among young and brilliant researchers, cultivated scientists, industry personnals and talented student com-munities. The contents of the book have been designed in such a way that it is providing extensive coverage of new developments, state of the art technologies, current and future trends in biotechnology and fermentation. The reader will be introduced with basic and advanced methodologies on industrial microbiology and fermentation technology. The main goal of this book is to share and enhance the knowledge of each and every individual in the fermentation world.

Ram A VishwakarmaDirector, CSIR-IIIM

xix

About the EditorsDr. Saurabh Saran, PhD, is a Fermentation Scientist hav-ing experience in Industrial microbiology, Biotechnology and Fermentation Technology for more than 15 years. Dr. Saran has completed his PhD from Delhi University, India. Dr. Saran has got hands-on experience in work-ing both industries and academic. He has worked in the industries like Reliance Industries Ltd., India. Later he was appointed as a Research Professor at Republic of Korea, South Korea. He has also worked as a Coordinator at the Technology Based Incubator, Delhi University South Campus, Delhi, Inida. Presently, he is working as a Senior

Scientist, Fermentation technology division, CSIR-IIIM, Jammu, India. He has an expertise on the screening, isolation, production and scale up of Industrial Enzymes, Biochemicals & Biofuels. Expert in process development/engineering, scale up to 5L, 10L, 30L, 100, 300 L & 500L fermentation size, (batch, fed batch and continuation fermentation) strain engi-neering, downstream processing and applications of industrially important biomolecules. To my credentials, I have 3 patents and more than 25 international publications in peer reviewed international journals on fermentation technology.

Dr. Vikash Babu, PhD was born in Bulandshahr dis-trict of Uttar Pradesh, India on 1st September 1981. He did his Bachelor’s degree from I.P (PG) College Bulandshahr, India. After qualifying all India combined entrance exam for biotechnology conducted by JNU, New Delhi, India, he did his degree in Biotechnology from Kumaun University, Nainital. After completing his M.Sc degree, he qualified many national level competi-tive exams such as DBT-JRF- 2005, CSIR-UGC NET for lecturership- Dec. 2004 & June 2005 and GATE-2005. In Nov. 2005, he joined as a DBT-JRF in the Department of Biotechnology, Indian Institute of Technology, Roorkee

under the superivision of Dr. Bijan Choudhury and registered for the Ph.D in the same department and Institute and completed his Ph.D degree in the year 2011. After finishing his Ph.D research work he joined Mangalayatan University, Beswan, Aligarh (India) as a lec-turer. He left Manglayatan University in the year 2012 and joined Graphic Era University, Dehradun (India) as an assistant professor where he worked till June 2013. Currently, he is working as a scientist in CSIR-IIIM.

xx About the Editors

Dr. Asha Chaubey, Ph.D is Principal Scientist and Head of Fermentation Technology Division, CSIR-Indian Institute of Integrative Medicine, Jammu, India. She has about 15 years of research experience in the area of enzymology and fermentation technology. She is actively engaged in development of indigenous process development. Her research interests include exploration and exploitation of microorganisms for production of enzymes and bioactives in special reference to industrial applications. She has published research articles in the area of bioactives production, enzyme immobilization, biotransformation, kinetic resolution of

racemic drug intermediates. She has also published several review articles and has been actively involved in the development of biosensors for health care and environmental moni-toring and has several patents on lactate and cholesterol biosensors.

xxi

List of Contributors

Akankszha Dwivedi, Biochemical Engineering Research and Process Development Centre CSIR- Institute of Microbial Technology, Chandigarh, (India)

Alok Malaviya, Department of Life Sciences, CHRIST (Deemed To Be University), Hosur Road, Bengaluru, (India)

Anil Agarwal, Department of Chemistry, CHRIST (Deemed To Be University), Hosur Road, Bengaluru, (India)

Anshela Koul, Fermentation Technology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu

Apurwa Kestwal, National Institute of Biologicals (Ministry of Health & Family Welfare) Government of India Plot No.A-32, Sector-62 Institutional Area, NOIDA, (U.P.), (India)

Aravind Madhavan, Rajiv Gandhi Center for Biotechnology, Thiruvananthapuram, Kerala, (India)

Asha Chaubey, Fermentation Technology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu Tawi, (India)

Ashok Pandey, CSIR- Indian Institute for Toxicology Research (CSIR-IITR), 31 MG Marg, Lucknow, (India)

Babbal, Adivitiya, Department of Microbiology, University of Delhi South Campus, New Delhi, (India)

Bhabatosh Chaudhuri, CYP Design Ltd, The Innovation Centre, 49 Oxford Street, Leicester, LE1 5XY, (UK)

Bhumica Agarwal, Department of Biotechnology, Meerut Institute of Engineering and Technology, Meerut, (India)

Cristóbal N. Aguilar Group of Bioprocesses. Food Research Department, School of Chemistry, Universidad Autónoma de Coahuila, Saltillo, Coahuila, (México)

Debendra K. Sahoo, Biochemical Engineering Research and Process Development Centre CSIR- Institute of Microbial Technology, Chandigarh, (India)

Dignya Desai, Amity Institute of Biotechnology, Amity University Rajasthan, Jaipur, (India)Divya Gupta, Department of Life sciences, Uttarakhand Technical University, Dehradun,

(India)

Edgard Gnansounou Ecole Polytechnique Federale de Lausanne, ENAC GR-GN, GC A3, Station 18, CH-1015, Lausanne, (Switzerland)

Girijesh K. Patel, Department of Oncologic Sciences, University of South Alabama, Mobile, (USA)

xxii List of Contributors

Gurdeep Singh, Biochemical Engineering Research and Process Development Centre CSIR- Institute of Microbial Technology, Chandigarh, (India)

Harikrishna Reddy , High Value Chemicals, Breakthrough R & D, Reliance Industries Limited. Reliance Corporate Park. Thane Belapur Road, GhansoliNavi Mumbai, (India)

Himanshu Verma, Biochemical Engineering Research and Process Development Centre CSIR-Institute of Microbial Technology, Chandigarh, (India)

Hitesh Sharma, Fermentation Technology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu(India)

Ibidapo Stephen Williams, CYP Design Ltd, The Innovation Centre, 49 Oxford Street, Leicester, LE1 5XY, (UK)

Ivanoe García, Group of Bioprocesses. Food Research Department, School of Chemistry, Universidad Autónoma de Coahuila, Saltillo, Coahuila, (México)

Jayesh Varavadekar, High Value Chemicals, Breakthrough R & D, Reliance Industries Limited. Reliance Corporate Park. Thane Belapur Road, GhansoliNavi Mumbai, (India)

Jitender Nandal, Biochemical Engineering Research and Process Development Centre CSIR- Institute of Microbial Technology, Chandigarh, (India)

Kakoli Dutt, Department of Bioscience and Biotechnology, Banasthali Vidyapith, Rajasthan (India)

Kanti N. Mihooliya, Biochemical Engineering Research and Process Development Centre CSIR-Institute of Microbial Technology, Chandigarh, (India)

Karan Malhotra, National Centre for Biological Sciences, Tata Institute of Fundamental Research, GKVK Campus, Bellary Road, Bengaluru, (India)

Katherine Saikia, Department of Life Sciences, CHRIST (Deemed To Be University), Hosur Road, Bengaluru, (India)

KB Arun, Rajiv Gandhi Center for Biotechnology, Thiruvananthapuram, Kerala, (India)

Lalit Kumar Singh, Department of Biochemical Engineering, School of Chemical Technology, Harcourt Butler Technical University Kanpur, (India)

Leonardo Sepulveda, Group of Bioprocesses. Food Research Department, School of Chemistry, Universidad Autónoma de Coahuila, Saltillo, Coahuila, (México)

Liliana Londoño-Hernández, Group of Bioprocesses. Food Research Department, School of Chemistry, Universidad Autónoma de Coahuila, Saltillo, Coahuila, (México)

Lipsy Chopra, Biochemical Engineering Research and Process Development Centre CSIR- Institute of Microbial Technology, Chandigarh, (India)

M. Sudhakara Reddy, Department of Biotechnology, Thapar University, Patiala 147004 (India)

Mahendra Pal Singh, National Institute of Biologicals (Ministry of Health & Family Welfare) Government of India Plot No.A-32, Sector-62 Institutional Area, NOIDA, (U.P.), (India)

Manali Datta, Amity Institute of Biotechnology, Amity University Rajasthan, Jaipur, (India)Manmeet Ahuja, High Value Chemicals, Breakthrough R & D, Reliance Industries Limited.

Reliance Corporate Park. Thane Belapur Road, GhansoliNavi Mumbai, (India)

List of Contributors xxiii

Mansi Vora, High Value Chemicals, Breakthrough R & D, Reliance Industries Limited. Reliance Corporate Park. Thane Belapur Road, GhansoliNavi Mumbai, (India)

Maryam Faiyaz1, Department of Bioengineering, Integral University, Lucknow, Dasauli, (India)

Parameswaran Binod , Microbial Processes and Technology Division, CSIR-National Institute for Interdisciplinary Science and Technology, Thiruvananthapuram, Kerala (India)

Piyush Sethia, High Value Chemicals, Breakthrough R & D, Reliance Industries Limited. Reliance Corporate Park. Thane Belapur Road, GhansoliNavi Mumbai, (India)

Pradeep Kumar, Biochemical Engineering Research and Process Development Centre CSIR- Institute of Microbial Technology, Chandigarh, (India)

Puja Tandon, School of Environmental Science and Engineering, Shanghai Jiao Tong University, Shanghai,PR (China)

Rahul Vikram Singh, Fermentation Technology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu(India)

Ramita Taggar, Biochemical Engineering Research and Process Development Centre CSIR- Institute of Microbial Technology, Chandigarh, (India)

Ramón Larios-Cruz, Group of Bioprocesses. Food Research Department, School of Chemistry, Universidad Autónoma de Coahuila, Saltillo, Coahuila, (México)

Raúl Rodríguez-Herrera Group of Bioprocesses. Food Research Department, School of Chemistry, Universidad Autónoma de Coahuila, Saltillo, Coahuila, (México)

Raveendran Sindhu, Rajiv Gandhi Center for Biotechnology, Thiruvananthapuram, Kerala, (India)

Ravi S. Manhas , Fermentation Technology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu, (India)

Ricardo Gómez-García, Group of Bioprocesses. Food Research Department, School of Chemistry, Universidad Autónoma de Coahuila, Saltillo, Coahuila, (México)

Richa Sharma, Fermentation Technology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu, (India)

Richi V Mahajan , National Institute of Biologicals (Ministry of Health & Family Welfare) Government of India Plot No.A-32, Sector-62 Institutional Area, NOIDA, (U.P.), (India)

Ruchika Goyal, Department of Biotechnology, Graphic Era University, Dehradun, (India)

Sanjog Garyali, Key Laboratory of Synthetic Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, (China).

Saurabh Saran, Fermentation Technology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu Tawi, (India)

Shikha Gupta, Gujarat State Biotechnology Mission, Gandhinagar, (India)Shilpa Mohanty, Department of Microbiology, University of Delhi South Campus, New Delhi,

(India) Subhash Chand, National Institute of Biologicals (Ministry of Health & Family Welfare)

Government of India Plot No.A-32, Sector-62 Institutional Area, NOIDA, (U.P.), (India)Surinder Singh, National Institute of Biologicals (Ministry of Health & Family Welfare)

Government of India Plot No.A-32, Sector-62 Institutional Area, NOIDA, (U.P.), (India)Syed M. Waheed, Department of Biotechnology, Graphic Era University, Dehradun, (India)

xxiv List of Contributors

Vidhya Rangaswamy, High Value Chemicals, Breakthrough R & D, Reliance Industries Limited. Reliance Corporate Park. Thane Belapur Road, GhansoliNavi Mumbai, (India)

Vikash Babu, Fermentation Technology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu

Yogender Pal Khasa, Department of Microbiology, University of Delhi South Campus, New Delhi, (India)

Yong Wang, Key Laboratory of Synthetic Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, (China).

xxv

PrefaceA century after the pioneering work of Louis Pasteur, the science of microbiology has reached to its zenith. In a short span of time, modern biotechnology has grown up drasti-cally from a laboratory scale to a commercial level. Advances in Fermentation Technology have created a favorable niche for the development of fermentation based products to facilitate their applications and to provide a sustainable environment for mankind and to improve the quality of human life. The modern day biotechnology offers many opportuni-ties and effective techniques to address the human concerns in the areas of pharmaceuti-cals, diagnostics, polymers, textiles, aquaculture, forestry, chemicals, household products, environmental cleanup, food processing, feed and forensics etc.

The book entitled “High Value Fermentation Products” has been divided in different volumes namely, Human Health and Human Welfare. The Volume 1 of the book has 18 chapters focussed on basics to fermentation technology, antibiotics & immunosuppres-sants, antibodies, peptides & proteins, sugars & sugar alcohols and metabolic engineering derived products. The fist chapter entitled ‘Introduction, scope and significance of fermen-tation technology’ aims to provide the insights on the basics of fermentation technology to the readers. The second chapter on ‘Extraction of bioactive molecules through fermenta-tion and enzymatic assisted technologies’ elaborates the techniques involved in isolation of bioactive molecules and other chemo-enzymatic approaches for bioactives production. Third and fourth chapters compile the important antibiotics against Gram positive and Gram negative bacteria discovered so far. Fifth chapter emphasizes on the role of antifungal drugs in combating invasive fungal diseases. Sixth chapter provides the update on rapamy-cin production and its potential clinical implications. The seventh chapter on ‘Advances in production of therapeutics monoclonal antibodies’ highlights the methodologies involved in the production of monoclonal antibodies for therapeutics. The eighth and ninth chapters focus on the antimicrobial peptides of microbial origin and their mechanism of produc-tion. The chapter tenth provides the insight on the therapeutic enzymes for human disease management. Chapters eleven to thirteen focus on the strategies involved in the produc-tion of natural sweeteners erythritol, xylitol and trehalose. Chapter fourteen describes how production of yeast derived microsomal human CYP450 enzymes (Sacchrosomes) with high yields and activities are superior to other commercially available microsomal enzymes. Chapters fifteen and sixteen compile the methodologies for production of artemisinin and flavonoids respectively. The chapter seventeen provides the information on how advances in metabolic engineering of the carotenoids can be useful for production of astaxanthin. The last chapter of the volume 1 describes how fungal endophytes can be exploited as bio-factories for production of functional metabolites through metabolic engineering, with special reference to taxol production.

xxvi Preface

This book provides deep insights on the strategies decisive factors involved in the fermen-tation based high value products like antibiotics, enzymes and other therapeutic secondary metabolites in the area of human health. We are confident that this book will be useful to students, researchers, academicians, and industry professionals interested in studying fer-mentation technology.

Editors

xxvii

AcknowledgementThe Editors take this opportunity to gratefully acknowledge the assistance and contribution of the people who have faith in us in this undertaking for compiling of the Book “‘High Value Fermentation Products”-Volume 1 (Human Health).

We are in debt of Dr. Ram A. Vishwakarma, Director, CSIR-Indian Institute of Integrative Medicine, Jammu for his valuable and esteemed guidance to carry out this task. His scholar-ship and authorative knowledge has been a great source of motivation and inspiration.

First and foremost, it is not enough to express our gratitude in words to all the contribu-tors for devotion and providing excellent matter of chapters on time.

The help and support provided by Mr. Chand Ji Raina, Mr. R.K. Khajuria and Mrs. Urmila Jamwal, was important and we acknowledge all of them with sincere thanks.

We are also thankful to the students of Fermentation Technology Division, CSIR-IIIM for their sincere efforts, dedication and determination to achieve objectives for the comple-tion of this task in a given time.

Where emotions are involved, words cease to mean for our family members for the con-sistent motivation during the planning and edition of this book.

We avail the opportunity to express our heartiest thanks to ‘Almighty’ for pouring His care and blessings throughout and making this work a success.

Saurabh SaranVikash Babu

Asha Chaubey