2nd Edition

Biotechnology Secondary Metabolites

Edited By K.G Ramawat, J.M. Merillon Copyright 2007
    390 Pages
    by CRC Press

    390 Pages
    by CRC Press

    This book provides new information relating recent advances made in the field of plant secondary products. Besides the updation of chapters this edition also includes chapters on secondary metabolites of microorganisms (fungi and lichen).

    Preface
    List of Contributors
    1. Biotechnology for Medicinal Plants: Research Need
    J.M. Merillon and K.G. Ramawat
    I. Introduction
    II. Medicinal Plants Exploration
    III. Resources: Wild and Cultivated Plants
    IV. Role of Biotechnological Approaches
    Development of Seed Material for Domestication
    Improvement — Somaclonal Variation and Genetic
    Engineering
    Understanding Metabolic Pathways
    New Compounds/Derivatives Production
    Scaling-up Technology Through Bioreactors
    Plant Molecular Farming
    Metabolomics
    V. Conclusions
    References
    VI. Further Readings
    2. Secondary Plant Products in Nature
    K.G. Ramawat
    I. Introduction
    II. Alkaloids
    A. Definition
    B. Biosynthesis of Alkaloids
    C. Occurrence of Alkaloids
    D. Distribution in Animal Kingdom and Lower Plants
    E. Classifications of Alkaloids
    F. Biological Functions of Alkaloids
    III. Other Metabolites
    A. Plant Amines
    B. Non-protein Amino Acids in Plants
    C. Quinones
    D. Resins
    E. Steroids
    F. Phenylpropanoids
    G. Polyisoprene (Rubber)
    H. Phenolics
    I. Flavours and Colours
    J. Insecticides of Plant Origin
    IV. Conclusions
    References
    V. Further Readings
    3. Factors Affecting the Production of
    Secondary Metabolites
    K.G. Ramawat and Meeta Mathur
    I. Introduction
    II. Culture Conditions
    The Environment
    III. Culture Initiation
    IV. Time-course Study of Growth and
    Secondary Metabolite Production
    V. Optimization
    A. Physical Factors
    B. Effect of Nutrients
    Use of Elicitors
    VII. Selection of Clones
    A. Variability in Field-grown Materials
    B. State of Differentiation
    C. Producer/Non-producer Cells
    D. Plating and Selection
    VIII. Clonal Stability
    IX. Flow Cytometry
    X. Optimization and Selection
    XI. Selection Parameters
    XII. Assessment of Somaclonal Variation
    XIII. Prospects
    References
    XIV. Further Readings
    4. Production of Food Additives
    G.A. Ravishankar, N. Bhagyalakshmi and S. Ramachandra Rao
    I. Introduction
    II. Colours
    A. Anthocyanins
    B. Betalaines
    C. Crocin and Crocetins
    D. Capsaicin and Other Capsaicinoids
    III. Flavours
    A. Vanilla
    B. Garlic and Onion
    IV. Sweeteners
    A. Steviosides
    B. Thaumatin
    V. Safety Aspects
    References
    5. Production of Insecticides
    Amita Pal
    I. Introduction
    II. Botanical Insecticides
    A. Phytoecdysterones
    B. Azadirachtin
    C. Rotenoids
    D. Pyrethrins
    E. Nicotine and Anabasine
    F. Quassin
    G. Limonene
    H. Ryania
    I. Sabadilla
    X. Optimization and Selection
    XI. Selection Parameters
    XII. Assessment of Somaclonal Variation
    XIII. Prospects
    References
    XIV. Further Readings
    4. Production of Food Additives
    G.A. Ravishankar, N. Bhagyalakshmi and S. Ramachandra Rao
    I. Introduction
    II. Colours
    A. Anthocyanins
    B. Betalaines
    C. Crocin and Crocetins
    D. Capsaicin and Other Capsaicinoids
    III. Flavours
    A. Vanilla
    B. Garlic and Onion
    IV. Sweeteners
    A. Steviosides
    B. Thaumatin
    V. Safety Aspects
    References
    5. Production of Insecticides
    Amita Pal
    I. Introduction
    II. Botanical Insecticides
    A. Phytoecdysterones
    B. Azadirachtin
    C. Rotenoids
    D. Pyrethrins
    E. Nicotine and Anabasine
    F. Quassin
    G. Limonene
    H. Ryania
    I. Sabadilla
    VI. Accumulation of Alkaloids
    A. Uptake and Storage
    B. Permealization
    C. Site of Alkaloid Accumulation
    D. Phenylalkylamines (Ephedra Alkaloids)
    E. Pyridine Alkaloids
    F. Tropane Alkaloids
    G. Quinolizidine and Pyrrolizidine Alkaloids
    H. Isoquinoline Alkaloids
    I. Quinoline (Cinchona) Alkaloids
    J. Monoterpene Indole Alkaloids
    K. Purine Alkaloids
    L. Ruta Alkaloids
    VII. Conclusions
    VIII. Prospects
    References
    8. Production of Steroids and Saponins
    Anupama Wagle, G.D. Kelkar and M.R. Heble
    I. Introduction
    II. Saponins
    A. Dioscorea
    B. Agave
    C. Glycyrrhiza
    D. Aesculus
    E. Panax
    III. Cardiac Glycosides
    A. Digitalis
    B. Strophanthus
    C. Urginea
    IV. Other Steroids
    A. Withania somnifera
    B. Holarrhena
    C. Solanum
    V. Conclusions
    References
    9. Understanding the Regulatory Mechanism
    of Secondary Metabolite Production
    J.M. Merillon and K.G. Ramawat
    I. Introduction
    II. Plant Growth Regulators, Elicitors, Sugars and Signal
    Transduction
    III. Protein and RNA
    IV. Genetic Manipulations
    A. Choice of Gene
    B. Manipulation of Biosynthetic Pathways
    V. Conclusions
    References
    10. Production of Secondary Metabolites by Bioconversion
    Niesko Pras and Herman J. Woerdenbag
    I. Introduction
    II. General Principles of Bioconversion
    III. Systems Applied for Bioconversion
    A. Freely Suspended Plant Cells
    B. Immobilized Plant Cells
    C. Enzyme Preparations
    IV. Kinetics of Immobilized Systems
    V. Bioconversion of Water-insoluble Precursors
    VI. Bioconversion of Synthetic Precursors
    VII. New Developments
    VIII. Final Consideration
    References
    11. Genetic Transformation for Production of
    Secondary Metabolites
    Sumita Jha
    I. Introduction
    II. Genetic Manipulation of Natural-Product Pathways
    III. Genetic Transformation for Development of
    Transformed Organ Cultures using Agrobacterium
    A. Relationship between Cell Organization,
    Differentiation and Secondary Metabolism
    B. Organ Cultures
    C. Transformed Organ Cultures
    D. Agrobacterium and Transformation Process
    IV. Applications of Transformed Root Cultures
    V. Experimental Procedure for Production of
    Transformed Hairy Roots in Artemisia annua Linn.
    VI. Experimental Procedure for Production of Transformed
    Shooty Teratomas in Coleus forskohlii Briq.
    VII. Prospects
    Acknowledgement
    References
    VIII. Further Reading
    12. Large-scale Production in Bioreactors
    J.M. Merillon
    I. Introduction
    II. Plant Cell Physiology
    III. Plant Cell Culture Processes
    A. Introduction
    B. Batch Cultivation
    C. Multi-stage Batch Cultivation
    D. Feed-Batch Cultivation
    E. Continuous Cultivation
    F. Semicontinuous Cultivation
    G. Immobilized Cell Cultivation
    IV. Bioreactor System Design and Operation
    A. Sterilization
    B. Oxygen Supply
    C. Bioreactor Types
    V. Applications
    A. Production of Secondary Metabolites
    B. Organ Culture
    C. Hairy Root Culture
    D. Commercialization
    V. Conclusions
    References
    13. Production of Ergot Alkaloids from Claviceps
    Arun Kumar and R. Raj Bhansali
    I. Introduction
    II. Historical Background
    III. Natural Occurrence
    IV. Alkaloids
    A. Formation and Production
    B. Chemical Properties
    C. Physical Properties
    D. Pharmacological Properties
    E. Chemistry
    F. Biosynthesis
    V. Artificial Culture
    VI. Conclusion
    References
    14. Lichen Products
    Wanda Quilhot, Cecilia Rubio and Ernesto Fernandez
    I. Introduction
    II. Chemistry of Lichen Substances
    III. Role of Lichen Metabolites in the Lichen Thallus
    IV. Biological
    A. Activity of Lichen Extracts and Lichen Compounds
    B. Anticancer-Antiviral Activity
    C. Inhibitory Enzyme and Antioxidant Activity
    D. Analgesic, Anti-inflammatory,
    Antipyretic and Other Medicinal
    E. Antiprotozoa Activity
    F. Antifeedant Activity
    G. Phototoxicidal and Other Activity in Plants
    H. Herbicidal Activity
    I. Plant Growth Activities
    J. Perfumes
    V. The Future
    References
    15. Chinese Herbal Drug Industry: Past,
    Present and Future
    Lixin Zhang and Wei Jia
    I. Introduction
    II. The Development of the Chinese Herbal Drug Industry
    III. Modernization of TCM
    A. The Status of Medicinal Plant Resources in China
    B. Status of Chinese Prepared Medicinal Herbs
    C. Status of TCM Manufacturing Enterprises
    D. Status of Plant Extracts
    E. Status of Pre-clinic and Clinical Research
    IV. The Internationalization of TCM
    A. Open Dialogue and Mutual Understanding
    Among All Parties Involved in Health Care System
    B. Deciphering the Preventive Nature of TCM
    C. Respect and Protect Intellectual Property
    D. Safety Issues of TCM Need to be Seriously Evaluated
    E. The Small Scale and Limited Capability of
    Herbal Drug Enterprises
    V. Closing Remarks
    Acknowledgments
    References
    16. Secondary Metabolites in in vitro Cultures of Ruta
    graveolens L. and Ruta graveolens ssp. divaricata
    (Tenore) Gams
    Halina Ekiert and Franz Ch. Czygan
    I. Distribution and Chemistry of the Investigated Plants
    A. Ruta graveolens L. (garden rue = common rue)
    B. Ruta graveolens ssp. divaricata (Tenore) Gams
    II. Groups of Secondary Metabolites Investigated in vitro —
    Their Chemical Characteristics, Distribution in Plant
    Kingdom and Biological Properties
    A. Linear Furanocoumarins
    B. Phenolic Acids
    C. Arbutin
    III. Production of the Investigated Metabolites in
    Plant in vitro Cultures
    A. Linear Furanocoumarins
    B. Phenolic Acids
    C. Arbutin
    IV. Studies from Our Laboratory
    A. Linear Furanocoumarins and other Coumarin
    Compounds Isolated as Main Fractions from Biomass
    Cultured in vitro
    B. Accumulation of Linear Furanocoumarins
    C. Accumulation of Phenolic Acids
    D. Biotransformation of Hydroquinone into Arbutin
    V. Conclusions and Prospects
    VI. Experimental
    A. Origin of Cultures Under Study
    B. Initial Stationary Liquid Cultures
    C. Experimental Cultures
    Acknowledgement
    References
    17. Antimicrobial and Cytotoxic Activity of Extracts and
    Secondary Metabolites Obtained from Plants and
    Lichens of Patagonia Austral.
    R.D. Enriz, M.L. Freile, E. Correche and M.J. Gomez-Lechon
    I. Introduction
    A. Natural Products as Sources for Lead Structures
    B. Cell Culture Systems in Toxicity Testing
    II. Techniques, Material and Methods
    A. Plant Material
    B. Extraction and Isolation of Test Compounds
    C. Microorganisms and Media
    D. Antimicrobial Assays
    E. Antifungal in vivo Assay
    F. Antifungal Activity
    G. Application of the Test Compounds
    H. Acute Toxicity Test
    I. Toxicity of Berberine to Embryo-larval
    Stages of Bufo arenarum (H). Test Conditions
    III. In vitro/in vivo Antimicrobial Activity of Aqueous
    Extracts and of Berberine Isolated from Berberis
    heterophilla
    IV. Antibacterial, Cytotoxic and Apoptotic Effects on
    Hepatocytes of Secondary Metabolites Obtained from
    Lichens of Patagonia
    A. Antibacterial and Cytotoxic Activity
    B. Apoptotic Activity
    V. Concluding Remarks
    Acknowledgements
    References
    18. Tools and Techniques for the Study of
    Plant Tissue Culture
    K.G. Ramawat and Meeta Mathur
    I. Microscopy
    II. Micrometry
    III. Cell Counting by Haemocytometer
    IV. Autoradiography
    V. Spectrophotometry
    A. UV-VIS Spectrophotometry
    B. Fluorescence Spectrophotometry
    VI. Centrifugation
    A. Analytical Model
    B. Preparative Model
    C. Application
    D. Methods of Separation
    VII. Gel Filtration
    VIII. Chromatography
    A. Column Chromatography
    B. Paper Chromatography
    C. Thin-layer Chromatography
    D. Adsorption Chromatography
    E. lon-exchange Chromatography
    F. Affinity Chromatography
    G. Gas Liquid Chromatography (GLC)
    H. High Performance Liquid Chromatography (HPLC)
    IX. Further Readings
    19. Practicals
    K.G. Ramawat
    I. Callus and Cell Culture
    A. Initiation of Callus
    B. Initiation of Suspension Culture
    C. Fresh Weight and Dry Weight Determinations
    II. Cell Viability Determination using Fluorescein Diacetate
    III. Diosgenin Estimation using Colorimeter
    IV. Solasodine Quantification
    V. Indole Alkaloid Production by Catharanthus roseus
    VI. TLC of Opium Alkaloids
    VII. Other Alkaloids
    VIII. Qualitative Tests
    A. Demonstration of Presence of Sterols
    B. Leuco-anthocyanin Test
    C. Saponin Test
    D. Phenol Test
    E. Flavonoid Test
    IX. Further Readings
    X. Annexures
    Index

    Biography

    K.G Ramawat, J.M. Merillon