Industrial Biotechnology of Vitamins, Biopigments, and Antioxidants

Erick J. Vandamme is Emeritus Professor at the Department of Biochemical and Microbial Technology, Faculty Bioscience Engineering, Ghent University, Belgium. He has acted as director of this department for over 25 years. He was Visiting Professor at several universities in Europe, America, Asia, and Australia. Following his Ph.D. studies at Ghent University in molecular biology, fermentation science and industrial biotechnology, several postdoctoral positions led him to Oxford University, MIT Cambridge, and Queen Elisabeth College (now King's College), London. Professor Vandamme is (co)-author of over 400 research papers and review articles , (co-)edited 14 books and holds several patents. He received numerous scientific awards and is an Elected Fellow of the American Academy of Microbiology (USA) and of the Society for Industrial Microbiology and Biotechnology, and received three honorary doctorates. He is a member of the Royal Flemish Academy of Belgium for Science and the Arts . Jose L. Revuelta is Full Professor of Genetics, Chairman of the Metabolic Engineering Group, and Director of the New Generation Sequencing Laboratory at the University of Salamanca (Spain) since 2002. Upon receipt of his Ph.D. in 1981 at Leon University in Biological Sciences, he received a grant by the Juan March Foundation to perform postdoctoral training at The Scripps Clinic and Research Foundation (La Jolla, CA). Professor Revuelta is coauthor of more than 50 research papers and review articles in the field of vitamins biotechnology, genomics and chemogenomics of industrial microorganisms. He coauthored eight chapters in books related with the biotechnological production of vitamins and pigments and holds 21 patents related to vitamin B2 production.

• List of Contributors XIXPreface XXVII1 Vitamins, Biopigments, Antioxidants and Related Compounds: A Historical, Physiological and (Bio)technological Perspective 1Erick J. Vandamme and José L. Revuelta1.1 Historical Aspects of the Search for Vitamins 11.2 Vitamins: What’s in a Name 31.3 Physiological Functions of Vitamins and Related Compounds 61.4 Technical Functions of Vitamins and Related Compounds 81.5 Production and Application of Vitamins and Related Factors 81.6 Outlook 13References 13Part I Water-Soluble Vitamins 152 Industrial Production of Vitamin B2 by Microbial Fermentation 17José L. Revuelta, Rodrigo Ledesma-Amaro, and Alberto Jiménez2.1 Introduction and Historical Outline 172.2 Occurrence in Natural/Food Sources 172.3 Chemical and Physical Properties; Technical Functions 182.4 Assay Methods and Units 182.5 Biological Role of Flavins and Flavoproteins 192.6 Biotechnological Synthesis of Riboflavin 212.6.1 Riboflavin-Producing Microorganisms 212.6.2 Biosynthesis of Riboflavin 222.6.3 Regulation of the Biosynthesis of Riboflavin 252.7 Strain Development: Genetic Modifications, Molecular Genetics and Metabolic Engineering 262.8 Fermentation Process 312.9 Downstream Processing 322.10 Chemical Synthesis 332.11 Application and Economics 33References 333 Vitamin B3, Niacin 41Tek Chand Bhalla and Savitri3.1 Introduction 413.2 History 423.3 Occurrence in Nature/Food Sources 433.4 Chemical and Physical Properties 443.4.1 Chemical Properties 443.4.2 Physical Properties 443.5 Vitamin B3 Deficiency Disease (Pellagra) 453.6 Methods Used for Determination of Vitamin B3 463.6.1 Microbiological Methods 463.6.2 Chemical Methods 463.7 Synthesis 473.7.1 Chemical Process Used for Nicotinic Acid Production 473.7.2 Biosynthesis 493.7.2.1 Biological Processes Used for Nicotinic Acid Production 493.8 Downstream Processing of Nicotinic Acid 523.9 Reactive Extraction 533.10 Physiological Role of Vitamin B3 (Niacin) 533.10.1 Coenzyme in Metabolic Reactions 533.10.2 Therapeutic Molecule 563.10.2.1 Treatment of Pellagra 563.10.2.2 Treatment of Cardiovascular Diseases 573.10.2.3 Antihyperlipidemic Effect 573.10.2.4 Treatment of Hypercholesterolemia 573.10.2.5 Diabetes 583.10.2.6 Fibrinolysis 583.10.2.7 Treatment of Neurodegenerative Disorders 583.11 Safety of Niacin 593.12 Toxicity of Niacin 593.12.1 Hepatotoxicity 593.12.2 Vasodilation/Niacin Flush 593.12.3 Glucose Intolerance 603.13 Derivatives of Niacin 603.14 Application in Cosmetics, Food and Feed 613.15 Future Prospects 61References 614 Pantothenic Acid 67Jesus Gonzalez-Lopez, Luis Aliaga, Alejandro Gonzalez-Martinez, and Maria V. Martinez-Toledo4.1 Introduction and Historical Outline 674.2 Occurrence in Natural Food Sources and Requirements 714.3 Physiological Role as Vitamin or as Coenzyme 744.4 Chemical and Physical Properties 774.5 Assay Methods 794.6 Chemical and Biotechnological Synthesis 814.7 Application and Economics 92References 985 Folate: Relevance of Chemical and Microbial Production 103Maddalena Rossi, Stefano Raimondi, Luca Costantino, and Alberto Amaretti5.1 Introduction 1035.2 Folates: Chemical Properties and Occurrence in Food 1035.3 Biosynthesis 1055.4 Physiological Role 1065.5 Bioavailability and Dietary Supplements 1095.6 Chemical and Chemoenzymatic Synthesis of Folic Acid and Derivatives 1105.7 Intestinal Microbiota, Probiotics and Vitamins 1145.8 Folate Production by Lactic acid Bacteria 1155.9 Folate Production by Bifidobacteria 1175.10 Conclusions 120References 1246 Vitamin B12 – Physiology, Production and Application 129Janice Marie Sych, Christophe Lacroix, and Marc J.A. Stevens6.1 Introduction and Historical Outline 1296.2 Occurrence in Food and Other Natural Sources 1306.3 Physiological Role as a Vitamin or Coenzyme 1316.3.1 Absorption and Transport 1316.3.2 Metabolic Functions 1326.3.3 Main Causes and Prevalence of Deficiencies 1336.3.4 Diagnosis of Deficiencies 1346.4 Chemical and Physical Properties 1346.5 Assay Methods 1376.6 Biotechnological Synthesis 1406.6.1 Producing Microorganisms 1406.6.1.1 Propionibacteria (PAB) 1426.6.1.2 Pseudomonades 1436.6.2 Biosynthesis and Metabolic Regulation 1446.6.3 Engineering of B12 Production 1456.6.3.1 Propionibacteria 1456.6.3.2 Pseudomonades 1466.6.4 Fermentation Process 1466.6.4.1 Propionibacteria 1466.6.4.2 Pseudomonades 1486.7 Downstream Processing; Purification and Formulation 1496.8 Application and Economics 1506.9 Conclusions and Outlook 151References 1517 Industrial Fermentation of Vitamin C 161Weichao Yang and Hui Xu7.1 Introduction and Historical Outline 1617.2 Occurrence in Natural/Food Sources 1627.2.1 Occurrence of Asc in Foods 1627.2.2 Biosynthesis of Asc in Plants and Mammals 1647.3 Physiological Role of Asc 1647.4 Chemical and Physical Properties 1657.5 Assay Methods 1657.6 Industrial Fermentation of Asc 1667.6.1 The Reichstein Process:The Major Industrial Asc Process until the Late 1990s 1677.6.1.1 The Establishment of the Reichstein Process 1677.6.1.2 Bioconversion of D-Sorbitol to L-Sorbose by Gluconobacter 1677.6.1.3 The Key Enzyme of Gluconobacter for L-Sorbose Production 1687.6.1.4 Oxidation of L-Sorbose to 2-KLG and Rearrangement to Asc 1687.6.2 The Two-Step Fermentation Process for Asc Production 1687.6.2.1 The First Step of Fermentation: Conversion of D-Sorbitol to L-Sorbose 1697.6.2.2 The Second Step of Fermentation: Conversion of L-Sorbose to 2-Keto-L-Gulonic acid 1707.6.2.3 Strain Development: Genetic Modification, Molecular Genetics and Metabolic Engineering 1757.6.2.4 Fermentation Process 1777.6.2.5 Upstream and Downstream Processing 1817.7 Application and Economics 1827.8 Outlook 183References 1858 Direct Microbial Routes to Vitamin C Production 193Günter Pappenberger and Hans-Peter Hohmann8.1 Introduction and Scope 1938.2 Principles of Direct L-Ascorbic Acid Formation:The Major Challenges 1958.2.1 Stereochemistry of L-Ascorbic Acid 1958.2.2 Enzymes Producing L-Ascorbic Acid and Their By-Product Spectrum 1968.3 Direct L-Ascorbic Acid Formation via 1,4-Lactones 1978.3.1 L-Ascorbic Acid Forming Enzymes: 1,4-Lactone Oxidoreductases 1988.3.2 Direct L-Ascorbic Acid Formation in HeterotrophicMicroalgae 2008.3.3 Direct L-Ascorbic Acid Formation in Recombinant Yeast 2018.3.4 Direct L-Ascorbic Acid Formation from Orange Processing Waste in Recombinant Aspergillus niger 2038.3.5 Overall Conclusion on 1,4-Lactone Routes 2048.4 Direct L-Ascorbic Acid Formation via 2-Keto Aldoses 2068.4.1 L-Ascorbic Acid Forming Enzymes: L-Sorbosone Dehydrogenases 2088.4.1.1 Sndhak 2088.4.1.2 Sndhai 2118.4.1.3 Prevalence of L-Asc Forming Sorbosone Dehydrogenases in Nature 2118.4.2 L-Asc or 2-KGA from L-Sorbosone: One Substrate, Several Isomers, Two Products 2128.4.3 L-Sorbose Dehydrogenase, Accumulating L-Sorbosone 2158.4.3.1 Ssdh from K. vulgare 2158.4.3.2 Sorbose Dehydrogenase Sdh from G. oxydans 2178.4.4 Gluconobacter as Host for Direct L-Ascorbic Acid Formation 2178.5 Outlook 219Acknowledgement 220References 220Part II Fat Soluble Vitamins 2279 Synthesis of ��-Carotene and Other Important Carotenoids with Bacteria 229Christoph Albermann and Holger Beuttler9.1 Introduction 2299.2 Carotenoids: Chemical Properties, Nomenclature and Analytics 2309.2.1 Nomenclature 2319.2.2 Analysis of Carotenoids 2319.2.2.1 Handling Precautions 2319.2.2.2 Extraction 2329.2.2.3 Chromatography Methods for Analysis of Carotenoids 2339.3 Natural Occurrence in Bacteria 2349.4 Biosynthesis of Carotenoids in Bacteria 2369.5 Biotechnological Synthesis of Carotenoids by Carotenogenic and Non-Carotenogenic Bacteria 2399.5.1 Heterologous Expression of Carotenoid Biosynthesis Genes 2409.5.2 Increased Isoprenoid Precursor Supply 2439.5.3 Genome-Wide Modification of E. coli to Increase Carotenoid Formation 2449.5.4 Balancing Recombinant Enzyme Activities for an Improved Synthesis of Carotenoids by E. coli 2499.5.5 Production of Industrially Important Carotenoids by Other Recombinant Bacteria 2529.5.6 Culture Conditions of Improved Formation of Carotenoids by Recombinant Bacteria 2529.6 Conclusion 253References 25410 ��-Carotene and Other Carotenoids and Pigments from Microalgae 265Borhane Samir Grama, Antoine Delhaye, Spiros N. Agathos, and Clayton Jeffryes10.1 Introduction and Historical Outline 26510.2 Occurrence in Nature and Food Sources 26610.3 Physiological Role as a Vitamin or as a Coenzyme 26710.4 Chemical and Physical Properties; Technical Functions 26810.5 Assay Methods and Units 27010.6 Biotechnological Synthesis 27010.6.1 Producing Organisms 27010.6.2 Biosynthesis and Metabolic Regulation 27310.6.3 Strain Development: Genetic Modification, Molecular Genetics and Metabolic Engineering 27610.6.4 Downstream Processing, Purification and Formulation 27610.7 Chemical Synthesis or Extraction 27910.8 Process Economics 279References 28011 Microbial Production of Vitamin F and Other Polyunsaturated Fatty Acids 287Colin RatledgeLipid Nomenclature 28711.1 Introduction: Essential Fatty Acids 28811.2 General Principles for the Accumulation of Oils and Fats in Microorganisms 29411.3 Production of Microbial Oils 29711.3.1 Production of Gamma-Linolenic Acid (GLA; 18 : 3 n-6) 29711.3.2 Productions of Docosahexaenoic Acid (DHA) and Arachidonic Acid (ARA) 30011.3.3 Alternative Sources of DHA 30211.3.4 Production of Eicosapentaenoic Acid (EPA n-3) 30511.3.5 Prospects of Photosynthetic Microalgae for Production of PUFAs 30711.4 Safety Issues 31011.5 Future Prospects 312Acknowledgements 315References 31612 Vitamin Q10: Property, Production and Application 321Joong K. Kim, Eun J. Kim, and Hyun Y. Jung12.1 Background of Vitamin Q10 32112.1.1 Historical Aspects 32112.1.2 Definition 32112.1.3 Occurrence 32212.1.3.1 In Nature 32212.1.3.2 In Food Sources 32212.1.3.3 In Microorganisms 32612.1.4 Functions 32612.2 Chemical and Physical Properties of CoQ10 32612.2.1 Chemical Properties 32612.2.2 Physical Properties 32712.3 Biosynthesis and Metabolic Regulation of CoQ10 32712.3.1 Biosynthesis of CoQ10 32712.3.1.1 Microorganisms 32712.3.1.2 Biosynthetic Pathways 32912.3.2 Metabolic Regulation 33412.3.3 Strain Development 33512.3.3.1 Mutagenesis 33512.3.3.2 Genetic Modification 33512.3.3.3 Metabolic Engineering 33712.3.4 Fermentation Process 33912.3.5 Upstream and Downstream Processing 34012.3.5.1 Upstream Processing 34012.3.5.2 Downstream Processing 34312.4 Chemical Synthesis and Separation of CoQ10 34512.4.1 Chemical Synthesis 34512.4.2 Solvent Extraction 34612.4.3 Purification 35012.5 Applications and Economics of CoQ10 35112.5.1 Applications 35112.5.1.1 In Diseases 35112.5.1.2 In Cosmetics 35212.5.1.3 In Foods and Others 35312.5.2 Economics 354References 35513 Pyrroloquinoline Quinone (PQQ) 367Hirohide Toyama13.1 Introduction and Historical Outline 36713.2 Occurrence in Natural/Food Sources 36713.3 Physiological Role as Vitamin or as Bioactive Substance 36813.4 Physiological Role as a Cofactor 37313.5 Chemical and Physical Properties; Technical Functions 37613.6 Assay Methods 37713.7 Biotechnological Synthesis 37713.7.1 Producing Microorganisms 37713.7.2 Biosynthesis and Metabolic Regulation 37813.8 Strain Development: Genetic Modification, Molecular Genetics and Metabolic Engineering 37813.9 Up- and Down-stream Processing; Purification and Formulation 38013.10 Chemical Synthesis or Extraction Technology 38013.11 Application and Economics 380References 381Part III Other Growth Factors, Biopigments and Antioxidants 38914 L-Carnitine, the Vitamin BT: Uses and Production by the Secondary Metabolism of Bacteria 391Vicente Bernal, Paula Arense, and Manuel Cánovas14.1 Introduction and Historical Outline 39114.2 Occurrence in Natural/Food Sources 39214.3 Physiological Role as Vitamin or as Coenzyme 39314.3.1 Physiological Role of Carnitine in the Mitochondria 39314.3.2 Physiological Role of Carnitine in the Peroxisomes 39414.3.3 Other Functions of Carnitine 39414.4 Chemical and Physical Properties 39414.5 Assay Methods and Units 39514.5.1 Chromatographic Methods 39514.5.2 MS-Based Methods 39514.5.3 Enzymatic Methods 39814.5.4 Automated Methods 39914.6 Biotechnological Synthesis of L-Carnitine Microbial Metabolism of L-Carnitine and Its Regulation 39914.6.1 Biotechnological Methods for L-Carnitine Production 39914.6.1.1 De novo Biosynthesis of L-Carnitine 39914.6.1.2 Biological Resolution of Racemic Mixtures 39914.6.1.3 Biotransformation from Non-Chiral Substrates 40014.6.2 Roles of L-Carnitine in Microorganisms 40114.6.2.1 Protectant Agent 40114.6.2.2 Carbon and Nitrogen Source 40114.6.2.3 Electron Acceptor: Carnitine Respiration 40214.6.3 L-Carnitine Metabolism in Enterobacteria and Its Regulation 40314.6.3.1 Metabolism of L-Carnitine in E. coli 40314.6.3.2 Metabolism of L-Carnitine in Proteus sp. 40514.6.4 Expression of Metabolising Activities: Effect of Inducers, Oxygen and Substrates 40614.6.5 Biotransformation with D-Carnitine or Crotonobetaine as Substrates 40614.6.6 Transport Phenomena for L-Carnitine Production 40714.6.6.1 Membrane Permeabilisation 40714.6.6.2 Osmotic Stress Induction of Transporters 40814.6.6.3 Overexpression of the Transporter caiT 40814.6.7 Metabolic Engineering for High-Yielding L-Carnitine Producing Strains 40814.6.7.1 Link between Central and Secondary Metabolism during Biotransformation 40814.6.7.2 Metabolic Engineering for Strain Engineering: Feedback between Modelling and Experimental Analysis of Cell Metabolism 40914.7 Other Methods for L-Carnitine Production: Extraction from Natural Sources and Chemical Synthesis 41114.7.1 Isolation of L-Carnitine from Natural Sources 41114.7.2 Chemical Synthesis 411Acknowledgement 412References 41215 Application of Carnosine and Its Functionalised Derivatives 421Isabelle Chevalot, Elmira Arab-Tehrany, Eric Husson, and Christine Gerardin15.1 Introduction and Historical Outline 42115.2 Sources and Synthesis 42215.2.1 Occurrence in Natural/Food Sources 42215.2.2 Chemical Synthesis of Carnosine 42215.2.3 Enzymatic Synthesis of Carnosine 42315.3 Physico-Chemical and Biological Properties of Carnosine 42515.3.1 Physico-Chemical Properties 42515.3.2 Physiological Properties 42615.4 Biotechnological Synthesis of Carnosine Derivatives: Modification, Vectorisation and Functionalisation 42715.4.1 Chemical Functionalisation 42715.4.2 Enzymatic Functionalisation: Enzymatic N-Acylation of Carnosine 43015.4.2.1 Lipase-Catalysed N-Acylation of Carnosine in Non-Aqueous Medium 43115.4.2.2 Acyltransferase-Catalysed N-Acylation of Carnosine in Aqueous Medium 43215.4.2.3 Impact of Enzymatic Oleylation of Carnosine on Some Biological Properties 43415.4.3 Vectorisation 43415.5 Applications of Carnosine and Its Derivatives 43515.5.1 Nutraceutics and Food Supplementation 43515.5.2 Cosmetics 43615.5.3 Pharmaceuticals 436References 43816 Metabolism and Biotechnological Production of Gamma-Aminobutyric Acid (GABA) 445Feng Shi, Yalan Ni, and Nannan Wang16.1 Introduction 44516.2 Properties and Occurrence of GABA in Natural Sources 44616.3 Metabolism of GABA 44716.3.1 Biosynthesis and Export of GABA 45016.3.1.1 Biosynthesis of GABA 45016.3.1.2 Essential Enzyme for GABA Biosynthesis – GAD 45116.3.1.3 Export of GABA 45216.3.2 Uptake and Catabolism of GABA 45416.3.2.1 The Uptake System of GABA 45416.3.2.2 The Catabolism of GABA 45516.4 Regulation of GABA Biosynthesis 45616.5 Biotechnological Production of GABA 45716.5.1 Fermentative Production of GABA by LAB 45816.5.2 Production of GABA by Enzymatic Conversion 45916.5.2.1 Production of GABA by Immobilised GAD 45916.5.2.2 Improving GAD Activity by Rational and Irrational Designs 45916.5.3 Fermentation of GABA by Recombinant C. glutamicum 46016.6 Physiological Functions and Applications of GABA 46116.6.1 Physiological Functions of GABA 46116.6.2 Applications of GABA 46216.7 Conclusion 462Acknowledgement 462References 46317 Flavonoids: Functions,Metabolism and Biotechnology 469Celestino Santos-Buelga and Ana M. González-Paramás17.1 Introduction 46917.2 Structure and Occurrence in Food 47117.3 Activity and Metabolism 47617.4 Biosynthesis of Flavonoids in Plants 48117.5 Biotechnological Production 48417.5.1 Reconstruction of Flavonoid Pathways in Plant Systems 48517.5.2 Reconstruction of Flavonoid Pathways in Microbial Systems 48717.5.2.1 E. coli Platform 48717.5.2.2 Saccharomyces cerevisiae Platform 48917.6 Concluding Remarks 489References 49018 Monascus Pigments 497Yanli Feng, Yanchun Shao, Youxiang Zhou,Wanping Chen, and Fusheng Chen18.1 Introduction and History of Monascus Pigments 49718.2 Categories of MPs 49718.3 Physiological Functions of MPs 49818.3.1 Anti-Cancer Activities 49818.3.2 Antimicrobial Activities 50818.3.3 Anti-Obesity Activities 50918.3.4 Anti-Inflammation Activities 51018.3.5 Regulation of Cholesterol Levels 51018.3.6 Anti-Diabetes Activities 51118.4 Chemical and Physical Properties of MPs 51118.4.1 Solubility 51118.4.2 Stability 51118.4.2.1 Effects of Temperature, pH and Solvent on Stability of MPs 51118.4.2.2 Effect of Light on Stability of MPs 51218.4.2.3 Effect of Metal Ion on Stability of MPs 51318.4.3 Safety 51318.5 Assay Methods and Units of MPs 51318.5.1 Extraction and Detection of MPs 51318.5.2 Isolation and Purification of MPs Components 51418.5.2.1 CC and TLC 51418.5.2.2 HPLC 51518.5.2.3 CE and the Others 51518.5.3 Identification of MPs Components 51518.6 MPs Producer – Monascus spp. 52018.6.1 Brief Introduction of Monascus Species and Their Applications 52018.6.2 Producing Methods of MPs 52018.6.3 Progress of Monascus spp. at the Genetic Level 52118.6.3.1 DNA Transformation 52118.6.3.2 Citrinin Synthesis and Its Regulations 52118.6.3.3 MK Synthesis and Its Regulations 52218.6.3.4 MPs Synthesis and Its Regulation 52218.6.3.5 The Regulation of Secondary Metabolism in Monascus spp. 52318.6.4 Monascus Genomics 52418.7 Application and Economics of MPs 524Acknowledgements 524References 526Index 537