Plants as Bioreactors for Industrial Molecules

Santosh Kumar Upadhyay is Assistant Professor in the Department of Botany at Panjab University in Chandigarh, India. He works in the area of plant molecular biology for the isolation, characterization, and recombinant production of various defense-related and industrial proteins. Sudhir P. Singh is a scientist of biotechnology and synthetic biology at the Center of Innovative and Applied Bioprocessing in Mohali, India. He works in the area of gene mining and biocatalyst engineering.

• About the Editors xvList of Contributors xviiPreface xxiiiAcknowledgments xxv1 Plants as Bioreactors: An Overview 1Madhu, Alok Sharma, Amandeep Kaur, Deepika Antil, Sudhir P. Singh, and Santosh Kumar Upadhyay1.1 Introduction 11.2 Factors Controlling the Production of Recombinant Protein 21.2.1 Choice of the Host Species 21.2.2 Optimization of Expression of Recombinant Protein 31.2.2.1 Transcription 41.2.2.2 Post- Transcription Modifications 61.2.2.3 Translation 71.2.2.4 Posttranslational Modifications (PTMs) of Recombinant Proteins 81.2.3 Downstream Processing 81.3 Recombinant Proteins in Plants 91.3.1 Pharmaceutical Proteins 91.3.2 Vaccine Antigens 131.3.3 Antibodies 141.3.4 Nutritional Molecules 151.3.5 Other Valuable Products 161.4 Conclusions 17References 172 Molecular Farming for the Production of Pharmaceutical Proteins in Plants 29Gaurav Augustine, Pragati Misra, Archana Shukla, Ghanshyam Pandey, and Pradeep Kumar Shukla2.1 Introduction 292.2 Plant as an Expression Platform 302.3 Plant- Derived Recombinant Proteins 342.4 Engineering Strategies Utilized for Recombinant Pharmaceutical Protein Production in Plants 342.4.1 Nuclear Transformation 352.4.2 Chloroplast Transformation 372.5 Pharmaceutical Protein Developed Using Plant Expression Platform 372.6 Perspectives 462.7 Conclusion 47References 473 Plants as Edible Vaccine 57Jia Qi Yip, Jia Choo, Kirthikah Kadiresen, Megan Min Tse Yew, Ying Pei Wong, and Anna Pick Kiong Ling3.1 Introduction 573.2 Mechanism of Action 593.3 Edible Plant Vaccines 603.3.1 Candidate Plants and Selection of Desired Gene 603.4 Production of Edible Vaccine (Plant Transformation) 613.4.1 Chemical- Mediated DNA Transfer Method 613.4.1.1 Polyethylene Glycol (PEG)- Mediated DNA Transfer Method 623.4.1.2 Liposome- Mediated DNA Transfer Method 623.4.1.3 Calcium Phosphate Coprecipitation 633.4.1.4 Diethylaminoethyl (DEAE) – Ddextran- mediated DNA Transfer Method 643.4.2 Direct Gene Delivery Method (Physical) 643.4.2.1 Biolistic Transfection 643.4.2.2 Electroporation 653.4.2.3 Sonication 653.4.2.4 Microinjection 663.4.3 Indirect Gene Delivery 663.4.3.1 Agrobacterium- Mediated Gene Transfer 663.4.3.2 Genetically Engineered Plant Virus 683.4.3.3 Virus- Like Particles (VLPs) 693.5 Plant Species Used as Vaccine Models 703.5.1 Potato 703.5.2 Rice 713.5.3 Banana 713.5.4 Tomato 723.5.5 Lettuce 723.5.6 Maize 733.5.7 Carrot 733.5.8 Alfalfa 733.6 Challenges 763.7 Conclusion 77Ackowledgments 77References 784 Plant Cell Culture for Biopharmaceuticals 89Zeuko’o Menkem Elisabeth and Rufin Marie Kouipou Toghueo4.1 Introduction 894.2 Plant Cultures 904.2.1 Plant Cell Cultures 904.2.2 Plant Tissue Culture 914.2.3 Plant Organ Cultures 924.3 Conditions for Plant Cell, Tissue, and Organ Culture 924.3.1 Culture Medium 924.3.2 pH 954.3.2.1 Plant Cell Growth Regulators (auxin, cytokinin, and gibberellin) 954.3.2.2 Auxins 954.3.2.3 Cytokinins 964.3.2.4 Gibberellins 964.3.2.5 Abscisic Acid (ABA) 964.4 Types of Plant Cell, Tissue, and Organ Culture 964.4.1 Embryo Culture 964.4.2 Somatic Embryogenesis 974.4.3 Genetic Transformation 974.4.4 Meristem Tip Culture 984.4.5 Organogenesis 984.4.6 Callus Culture (Callogenesis) 984.4.7 Adventitious Root/Hairy Root Culture (rhizogenesis) 984.4.8 Suspension Culture 994.4.9 Protoplast Fusion 994.4.10 Haploid Production 994.4.11 Germplasm Conservation 1004.5 The Techniques Used in Plant Culture 1004.5.1 Micropropagation in Medicinal Plants 1014.5.1.1 Stage 0: Preparation of the Donor Plant 1014.5.1.2 Stage I: Initiation Stage 1014.5.1.3 Stage II: Multiplication Stage 1024.5.1.4 Stage III: Rooting Stage 1024.5.1.5 Stage IV: Acclimatization Stage 1024.5.2 Elicitation 1024.5.3 Transformed Tissue Cultures 1034.5.4 Metabolic Engineering 1044.6 Applications of Plant Cultures 1044.7 Biopharmaceuticals 1044.7.1 Biopharmaceuticals from Plants 1054.7.1.1 Scale- up of Secondary Metabolites by Using Different Systems 1074.7.1.2 Vaccines 1104.7.1.3 Plantibodies 1154.7.1.4 Proteins 1154.7.2 The Effects of Production, Safety, and Efficacy 1184.8 Conclusion 118References 1195 Microalgal Bioreactors for Pharmaceuticals Production 127Rufin Marie Kouipou Toghueo5.1 Introduction 1275.2 Microalgae Strains Selection 1285.3 Microalgae Cultivation 1295.3.1 Factors Affecting the Growth and Productivity of Microalgae 1305.3.1.1 Nutrients 1305.3.1.2 Temperature 1315.3.1.3 pH, Salinity, and Pressure 1325.3.1.4 Light 1325.3.1.5 Mixing 1335.3.2 Methods and Systems for Microalgae Cultivation 1345.3.2.1 Methods 1345.3.2.2 Microalgae Cultivation Systems 1365.4 Acquiring Biopharmaceuticals from Microalgae’s 1375.4.1 Microalgae Harvesting 1375.4.1.1 Flocculation and Ultrasound 1385.4.1.2 Centrifugation 1385.4.1.3 Filtration 1385.4.1.4 Flotation 1395.4.2 Biomass Dehydratation 1395.4.3 Cell Disruption for Bioproducts Extraction 1405.5 Microalgal Compounds and their Pharmaceutical Applications 1415.5.1 Carotenoids 1415.5.2 Polyunsaturated Fatty Acids 1435.5.3 Polysaccharides, Vitamins, and Minerals 1455.5.4 Proteins 1455.6 Conclusions 147References 1476 Micropropagation for the Improved Production of Secondary Metabolites 161Rupasree Mukhopadhyay6.1 Introduction 1616.2 Micropropagation for Production of Secondary Metabolites 1636.3 Strategies to Improve Secondary Metabolite Production 1656.3.1 Optimizing Culture Conditions 1656.3.2 Selecting High- Producing Cell Lines 1676.3.3 Organ Cultures 1676.3.4 Precursor Feeding 1686.3.5 Elicitation 1686.3.6 Immobilization 1706.3.7 Permeabilization 1716.3.8 Genetic Transformation: Hairy Root Cultures and Shooty Teratomas 1716.3.9 Biotransformation 1726.3.10 Metabolic Engineering 1736.3.11 Plant Bioreactors and Scale- up 1746.4 Conclusions 176References 1767 Metabolic Engineering for Carotenoids Enrichment of Plants 185Monica Butnariu7.1 Background 1857.2 Classification of Carotenoid Pigments 1867.2.1 Carotenoid Hydrocarbons 1917.2.2 Xanthophylls 1927.2.3 Carotenoid Ketones 1927.2.4 Carotenoid Acids 1937.3 Aspects of the Mechanism of Carotenoid Biosynthesis 1947.3.1 Premises of Metabolic Engineering 2087.4 Concluding Remarks and Future Perspectives 209References 2108 Plant Genome Engineering for Improved Flavonoids Production 215Monica Butnariu8.1 Background 2158.2 Structure, Diversity, and Subgroups 2178.3 Flavonoid Biosynthesis 2238.4 The Mechanism of Action of Flavonoids 2298.5 The Role of Flavonoids in Food and Medicine 2338.6 Concluding Remarks and Future Perspectives 236References 2369 Antibody Production in Plants 241Vipin Kumar Singh , Prashant Kumar Singh , and Amit Kumar Mishra9.1 Introduction 2419.2 How Are Antigens Expressed in Plants? 2429.2.1 Transient Expression of Antigens 2429.2.2 Plant Virus Fusion Proteins 2439.3 Plant- Derived Antibodies: Are There any Alternative Approaches? 2449.4 Antibody Production in Plants: Advantages and Concerns 2469.5 Conclusion and Prospects 247References 24810 Metabolic Engineering of Essential Micronutrients in Plants to Ensure Food Security 255Swarnavo Chakraborty and Aryadeep Roychoudhury10.1 Introduction 25510.2 Metabolic Engineering of Crops for Increased Nutritional Value 25610.2.1 Iron 25610.2.2 Iodine 26010.2.3 Zinc 26010.2.4 Vitamin A 26110.2.5 Vitamin B 626310.2.6 Vitamin B 9 26410.2.7 Vitamin E 26510.3 Conclusion and Future Perspectives 266Acknowledgments 266References 26811 Plant Hairy Roots as Biofactory for the Production of Industrial Metabolites 273Nidhi Sonkar, Pradeep Kumar Shukla, and Pragati Misra11.1 Introduction 27311.2 Types of Metabolites and Industrial Metabolites 27411.3 Secondary Metabolites 27611.4 Importance of Secondary Metabolites 27711.5 Enhancement of Secondary Metabolites 27811.6 Hairy Roots 28011.6.1 Hairy Roots 28011.6.2 Hairy Roots in Plants and In vitro Production of Secondary Metabolites 28111.7 Initiation of Hairy Root Cultures 28211.7.1 Formation of Highly Proliferative Hairy Roots 28211.7.2 Agrobacterium rhizogenes for Hairy Root Production and as a Biotechnology Tools 28311.8 Large- Scale Production of Secondary Metabolites 28511.9 Strategies Used In vitro 28711.9.1 Why Hairy Root Culture? 28911.10 Plants as Bioreactors 28911.11 A Case Study 29111.12 Conclusion 292References 29412 Microalgae as Cell Factories for Biofuel and Bioenergetic Precursor Molecules 299D. Rodríguez- Zuñiga, A. Méndez- Zavala, O. Solís- Quiroz, J.C. Montañez, L. Morales- Oyervides, and J.R. Benavente- Valdés12.1 Introduction 29912.2 Microalgae that Produce Bioenergy and Biofuel Molecules 30012.3 Biosynthesis of Molecules for Bioenergy and Biofuels in Microalgae 30212.4 Biohydrogen Production 30312.5 Starch Biosynthesis 30312.6 Lipid Biosynthesis 30412.7 Biochemical Regulation of BBPM Associated with Nutritional Conditions 30612.8 Physical and Chemical Factors Promote the Accumulation of Molecules for Bioenergy and Biofuels 30812.9 Light Intensity 30812.10 Salts 30812.11 Use of Organic and Inorganic Carbon Sources 30912.12 Agitation 30912.13 Photobioreactors to Produce Bioenergy and Biofuels 31012.14 Open Pond Cultivation Systems 31012.15 Closed Systems 31012.16 Hybrid Systems 31112.17 Conclusions 311References 31113 Metabolic Engineering for Value Addition in Plant- Based Lipids/Fatty Acids 317Himani Thakkar and Vinnyfred Vincent13.1 Introduction 31713.2 Plant Lipids 31813.3 Tag Synthesis in Plants 31813.3.1 Fatty Acid Synthesis 31813.3.2 Tag Biosynthesis 31913.3.3 Lipid Droplets Biogenesis 32013.3.4 Wax Esters Synthesis 32113.4 Regulatory Factors Involved in Tag Synthesis 32213.5 Metabolic Engineering for Lipid/Fatty Acid Synthesis 32313.5.1 Increasing Oil Accumulation in Plants 32513.5.1.1 Modification of Fatty Acid Synthesis Pathway 32513.5.1.2 Increasing Tag Synthesis/Assembly Process 32513.5.1.3 Increasing Carbon Flux Toward Oil Biosynthesis 32513.5.1.4 Modulating the Expression of Transcription Factors 32613.5.1.5 Reducing the Hydrolysis of Storage Lipids 32613.5.2 Improving the Quality of Oil by Altering the Fatty Acid Profile 32613.6 Conclusions 327References 33114 Plants as Bioreactors for the Production of Biopesticides 337Fernanda Achimón, Vanessa A. Areco, Vanessa D. Brito, María L. Peschiutta, Carolina Merlo, Romina P. Pizzolitto, Julio A. Zygadlo, María P. Zunino, and Alejandra B. Omarini14.1 Introduction 33714.2 Plant Metabolic Engineering for the Production of EOs and their Pure Compounds 33814.3 Bioactivity of EOs 34114.3.1 Insecticidal Effects of EOs 34114.3.1.1 EO Composition of the Lamiaceae Main Genera with Insecticidal Effect 34114.3.1.2 Characteristics of Some Species Within the Main Genera 34214.3.2 Antibacterial Activity of EOs 34514.3.3 Antifungal Effect of EOs 34714.3.4 Bioconversion Process of EOs and Their Components by Microorganisms 35414.4 In vitro Synthesis vs Extraction from Natural Sources: How to Obtain Secondary Metabolites 35614.4.1 Factors Affecting the Extraction of Bioactive Compounds from Natural Sources 35614.4.2 Production of Azadirachtin by Azadirachta indica. A Case Study 35714.5 Conclusion 358References 35915 Nutraceuticals Productions from Plants 367Isabela Sandy Rosa, Laura Oliveira Pires, and Juliane Karine Ishida15.1 Plant- Derived Nutraceuticals 36715.2 Phytochemicals and their Impacts on Human Health 36915.2.1 Polyphenols 36915.2.1.1 Chromones 37015.2.1.2 Coumarins 37115.2.1.3 Flavonoids 37115.2.1.4 Curcumin 37315.2.1.5 Stilbenes 37315.2.1.6 Xanthones 37415.2.2 Terpenoids 37515.2.2.1 Carotenoids 37615.2.2.2 Ginkgolides 37615.2.2.3 Limonene 37615.2.2.4 Oleanolic Acid 37615.2.2.5 Phytosterols 37615.2.2.6 Tocopherols and Tocotrienols 37715.2.3 Alkaloids 37715.2.4 Fatty Acids 37915.2.5 Fiber 38015.3 Engineering Nutraceutical- Enriched Plants 38115.4 Potential Side Effects of Nutraceuticals on Human Health 38215.5 Final Considerations 383References 38416 Green Synthesis of Nanoparticles Using Various Plant Parts and Their Antifungal Activity 393Chikanshi Sharma, Madhu Kamle, and Pradeep Kumar16.1 Introduction 39316.2 Gold Nanoparticle Synthesis Using Plant Source 39516.3 Silver Nanoparticles Synthesis Using Plants Source 39916.4 Zinc Oxide Nanoparticles Synthesis Using Plants 40016.5 Other Nanoparticles Synthesis Using Plant Source 40116.6 Conclusion and Future Perspective 402Acknowledgement 402Conflicts of Interest 403Author Contribution 403References 40317 Plant- Based/Herbal Nanobiocatalysts and Their Applications 411Rajeswaree Gohel, Dhara Gandhi, and Gaurav Sanghvi17.1 Introduction of Nanobiocatalyst 41117.2 Nanobiocatalysts from Herbal Alkaloid Plants Are Used in Nanotechnology and Bioengineering 41217.3 Why Use Nanobiocatalysts? 41317.4 Immobilization of Biocatalyst (Enzymes) and Nanoparticles or Nanomatrix 41317.5 Application of the Nanobiocatalyst 41517.5.1 Application of Enzyme Immobilized on Graphene- Based Nanomaterial 41517.5.2 Enzyme- Based Biosensor 41517.5.2.1 Horseradish Peroxidase Immobilized with the Graphene Oxide (GO) 41617.5.2.2 HRP Biosensor Towards the Detection of Dopamine 41617.5.2.3 HRP – Inorganic Hybrid Nanoflower 41717.5.3 Bitter Gourd Peroxidase Immobilized with TiO 2 Nanoparticles 41717.5.4 Immobilization of Acetylcholinesterase on Gold Nanoparticles Embedded in Sol–Gel Nanomatrix 41817.5.5 Alcohol Dehydrogenase Immobilized with Carbon Nano Scaffold 41817.5.6 Vanillin or Vanillin Synthase is Used as a Therapeutic Drug by Immobilizing with Nanoparticles 41917.5.7 STR Gene Regulation with the Help of Silver Nanoparticles 41917.5.8 Effect of Titanium Dioxide Nanoparticles and Different Enzymes of Alkaloid Plants Conjugate on the Bioengineering Pathway 42017.5.9 Application of Plant Extract Biocatalyst Which is Useful to Make Different Nanoparticles and Used as a Remedy. See Table 17.2. 42117.6 Conclusion 422References 42218 Potential Plant Bioreactors 427Karishma Seem and Simardeeep Kaur18.1 Introduction 42718.2 Whole Plants: Stable and Transient Expression Systems 42918.2.1 Stable Expression (Whole Plant Based) 42918.2.1.1 Leaf Based 42918.2.1.2 Seed Based 43118.2.2 Transient Expression 43218.2.3 In vitro Culture Systems 43318.2.3.1 Plant Suspension Cultures 43418.2.3.2 Hairy Root System 43518.2.3.3 Moss 43818.2.4 Aquatic Plants 43818.2.4.1 Duckweed 43818.2.4.2 Microalgae 43918.3 Unique Features of Using Plant- based Production Over Microbial and Mammalian Systems 44118.3.1 Better Protein Functionality 44218.3.2 Plant Matrix 44218.3.3 Speed and Scalability of Production 44218.3.4 Consumer Acceptance 44218.3.5 Animal- free Production thus Lower Risks of Pathogen Invasion 44218.4 Strategies to Enhance the Potential of Plant- based Production Systems 44318.4.1 To Minimize Ecological Footprint via Inherent Carbon dioxide Fixation and Improved and Sustainable Fertilizer Use 44318.4.2 Use of Pant Bioreactors to Harvest Multiple Products from a Single Process 44318.4.3 Reduced Investment and Establishment of Vertical Farms 44418.4.4 Use of Biodegradable Plant- based Expression Systems 44518.5 Concluding Remarks and Future Perspectives 445Conflict of Interest 446References 44619 Production of Nutraceuticals Using Plant Cell and Tissue Culture 457Elif Karlik and Elif Aylin Ozudogru19.1 Introduction 45719.2 Production of Secondary Metabolites as Nutraceuticals in In vitro Cultures 45919.2.1 Nutraceuticals Used in Pharmaceuticals Industry 45919.2.2 Nutraceuticals Used in Food and/or Cosmetic Industry 46519.3 Conclusions 472References 47220 Algal Bioreactors for Polysaccharides Production 485Michele Greque de Morais, Priscilla Quenia Muniz Bezerra, Kricelle Mosquera Deamici, Suelen Goettems Kuntzler, Juliana Botelho Moreira, Céline Laroche, and Jorge Alberto Vieira Costa20.1 Introduction 48520.2 Algae 48620.2.1 Algae Producers of Polysaccharides 48620.2.2 Types of Algae Polysaccharides 48720.3 Biological Activity of Algal Polysaccharides 48820.4 Parameters that Iinfluence the Polysaccharides Production by Microalgae 48920.4.1 Chemical Parameters 49020.4.2 Physical Parameters 49120.5 Algal Bioreactors 49220.5.1 Open System 49320.5.2 Closed System 49420.6 Conclusions and Future Perspectives 494Acknowledgments 495References 495Index 503