Biotechnology of Natural Products / edited by Wilfried Schwab, Bernd Markus Lange, Matthias Wüst.

Preface
Contents
Contributors
Part I: Biotechnological Production of Selected Natural Products
1: Vanilla: The Most Popular Flavour
1.1 Introduction
1.2 Vanilla Orchids, Vanilla Flowers and the Pod
1.3 When the Green Becomes Black – Vanilla Pod Curing
1.4 Vanillin Is the Key Flavour Compound of the Complex Vanilla Extract
1.5 How Does the Vanilla Plant Form Vanillin?
1.6 Flavour Synthesis by Brewing – Bioengineering of Vanillin Biosynthesis
1.7 Biotechnology-Based Production of Vanillin from Eugenol, Iso-Eugenol, Ferulic Acid and Gluco
1.8 Future Perspectives and Final Remarks
References
2: Rosmarinic Acid and Related Metabolites
2.1 Occurrence and Structures of Rosmarinic Acid and Related Metabolites
2.2 Biosynthetic Pathway of Rosmarinic Acid
2.3 Production of RA in Untransformed Aseptic In Vitro Cultures
2.3.1 Species from the Family Lamiaceae
2.3.2 Species of the Family Boraginaceae
2.3.3 Non-vascular Plant Species
2.4 Production of Rosmarinic Acid in Hairy Roots
2.4.1 Hairy Roots of Lamiaceae Species
2.4.2 Hairy Roots of Boraginaceae Species
2.5 Production of Rosmarinic Acid and Related Caffeic Acid Esters in Microorganisms
2.6 In Vitro Formation of Non-natural Hydroxycinnamic Acid Esters and Amides by “Rosmarinic Ac
2.7 Conclusion and Outlook
References
3: Bioproduction of Resveratrol
3.1 Introduction
3.2 Biosynthesis of Resveratrol and Its Analogs
3.3 Bioproduction of Resveratrol in Microorganisms
3.3.1 Yeast
3.3.2 E. coli
3.3.3 Other Bacteria
3.4 Bioproduction of Resveratrol Analogs and Derivatives
3.5 Strategies in Metabolic Engineering of Resveratrol Production
3.5.1 Pathway Engineering to Increase Precursor Supply
3.5.2 Protein Engineering
3.6 Conclusion and Perspective
References
4: Anthocyanin Production in Engineered Microorganisms
4.1 Anthocyanins and Their Industrial Applications
4.1.1 Pharmaceutical Applications
4.1.2 Food Colorants
4.1.3 Cosmetic Industry
4.1.4 Other Fields
4.2 Plant-Based Anthocyanin Production
4.2.1 Extraction from Plants
4.2.2 Anthocyanin Production from Suspension Cell Culture
4.3 Anthocyanin Production in Microorganisms
4.3.1 Engineering of Pathway Enzymes
4.3.2 Supply of Cofactors and Cosubstrates
4.3.3 Engineering Anthocyanin Secretion
4.3.4 Optimization of the Production Process
4.4 Conclusions and Future Perspectives
References
5: Microbial Synthesis of Plant Alkaloids
5.1 Introduction
5.2 Reconstitution of MIA Biosynthetic Pathways
5.2.1 Precursor Pathways
5.2.2 Strictosidine Formation
5.2.3 Downstream Derivatization
5.3 Reconstitution of BIA Biosynthetic Pathways
5.3.1 Upstream BIA Pathways
5.3.2 Microbial Production of Morphinan Alkaloids
5.3.3 Microbial Production of Protoberberine, Benzophenanthridine, and Phthalideisoquinoline Alka
5.3.3.1 Synthesis of Dihydrosanguinarine and Sanguinarine in Yeast
5.3.3.2 Noscapine Synthesis in Yeast
5.4 Microbial Engineering Challenges
5.4.1 Functional Expression of Cytochromes P450 (CYPs)
5.4.2 Taming Enzyme Promiscuity
5.4.3 Limiting Efflux of Intermediates
5.5 Optimization Strategies
5.5.1 Genetic and Pathway Engineering Techniques
5.5.1.1 Building Combinatorial Enzyme Libraries
5.5.1.2 Pathway Assembly
5.5.1.3 Tuning Gene Expression
5.5.2 Cultivation Methods
5.6 Conclusions and Future Directions
References
6: Caffeine
6.1 Introduction
6.2 Distribution of Caffeine in Plants
6.3 Biosynthesis of Caffeine from Xanthosine
6.4 The Caffeine Synthase Gene Family in Plants
6.5 Biotechnological Production of Caffeine by Genetic Engineering
References
7: Taxol® Biosynthesis and Production: From Forests to Fermenters
7.1 Introduction
7.2 The Biosynthesis of Taxol
7.3 Taxol Production by Endophytes
7.4 Taxol Production by Plant Cell Cultures
7.5 Microbial Biotechnology for the Production of Taxol
7.6 Concluding Remarks
References
Part II: Technologies for Metabolic, Enzyme and Process Engineering
8: Commercial-Scale Tissue Culture for the Production of Plant Natural Products: Successes, Failu
8.1 Introduction
8.2 Commercial Products from Plant Tissue Culture
8.3 Considerations for Commercial Targets
8.4 Future Opportunities
References
9: Tailoring Natural Products with Glycosyltransferases
9.1 Introduction
9.2 The Significance of Glycosylation in Plants
9.2.1 Glycosylation Increases Solubility
9.2.2 Glycosylation Increases Stability
9.2.3 Glycosylation Controls Sequestration/Compartmentalization
9.2.4 Glycosylation Affects Bioactivity and -Availability
9.2.5 Glycosylation Reduces Toxicity
9.2.6 Glycosylation Affects Perception
9.3 Glucoside/Glucose Ester Synthesis
9.4 Family 1 Plant Glycosyltransferases
9.5 Substrates of Family 1 Plant Glycosyltransferases
9.5.1 Secondary Metabolites
9.5.1.1 Phenylpropanoids
9.5.1.2 Flavonoids, Anthocyanins
9.5.1.3 Dihydrochalcones, Acylphloroglucinol, Stilbenes, Curcumin
9.5.1.4 Terpenoids
9.5.2 Plant Hormones
9.5.3 Miscellaneous Substrates (Alkaloids, Benzoxazinoids, Furanones, and Xenobiotics)
9.6 Glucoside Production by Whole Cell Biocatalysts
9.6.1 Production System
9.6.2 Types of Whole-Cell Biocatalysts
9.6.3 Process Optimization
9.6.3.1 Vector Conveyed Process Optimization
9.6.3.2 UGT Optimization
9.6.3.3 Host Genome Optimization – Metabolic Engineering
9.7 Recent Applications of Glycosyltransferases for Production of Small Molecule Glycosides
9.7.1 Glycosyltransferases and Glycosylation in Product for Consumer Consumption
9.7.2 Glycorandomization
9.7.3 Glycosyltransferases and Their Role in Cancer Therapies
9.8 Conclusions and Future Prospects
References
Part III: Analytical and Legal Aspects
10: Authenticity Control of Natural Products by Stable Isotope Ratio Analysis
10.1 Introduction
10.2 Isotope Ratio Mass Spectrometry – IRMS
10.2.1 Notations in IRMS
10.2.2 Elemental Analyzer-IRMS (EA-IRMS) and Gaschromatography-IRMS (GC-IRMS)
10.2.3 Liquid Chromatography-IRMS (LC-IRMS)
10.2.4 Site-Specific Natural Isotope Fractionation-Nuclear Magnetic Resonance Spectroscopy (SNIF-NM
10.3 Practical Applications of Stable Isotope Analysis on Miscellaneous Natural Products
10.3.1 Phenylpropanoids (Vanillin)
10.3.2 Terpenes (Monoterpenes and Tetraterpenes)
10.3.3 Polyphenols (Resveratrol)
10.3.4 Alkaloids (Caffeine)
10.4 Conclusions
References
11: Natural or Synthetic? The Legal Framework in the EU for the Production of Natural Flavouri
11.1 Introduction
11.2 General Conditions
11.3 Scope of the Regulation
11.4 Definitions
11.4.1 Definitions of Flavourings in General
11.4.2 Definition of Natural Flavouring Substance
11.4.3 Definitions of Flavouring Preparations
11.5 Processes for the Production of Natural Flavouring Ingredients
11.5.1 EFFA Considerations on the Permitted Order of the Various Processes
11.5.2 Biotechnology for the Production of Natural Flavouring Ingredients
11.5.3 Further Considerations on the Production of Natural Flavouring Substances and Flavouring
11.6 Analytical Methods to Assess Authenticity
11.7 Labelling of Flavourings (B2B)
11.7.1 General Labelling Requirements
11.7.2 Labelling of Natural Flavourings
11.7.3 Considerations on Different Interpretations of the 95/5-­Rule (Art. 16(4))
11.8 Safety Evaluation of Flavourings and Their Inclusion in the EU Union List
References
Index

This text comprehensively covers the analysis, enzymology, physiology and genetics of valuable natural products used in the food industry that are attractive targets for biotechnological production. The focus is on the recent advances made to achieve this goal. This unique work is the first book to focus on biotechnological production of important natural products in food additives, fragrances and flavorings, and other bioactive compounds in food. The chapters offer a deep insight into modern research and the development of low molecular weight natural products. Biotechnology of Natural Products covers products in the Phenolic, Terpenoid, and Alkaloid categories, providing a full overview of the biotechnology of food additives and other low molecular weight natural products. Gene clustering and the evolution of pathways are covered, as well as future perspectives on the topic. Due to limited oil resources and increasing consumer demand for naturalness, bioprocesses are increasingly needed to meet these requirements. Novel sophisticated technologies have facilitated the elucidation of new chemical molecules, their biosynthetic pathways and biological functions. This book provides researchers with a full overview of the technologies and processes involved in the biotechnology of natural products.