Handbook of Vanilla Science and Technology

Dr. Daphna Havkin-Frenkel, Department of Plant Biology, School of Environmental and Biological Sciences, Rutgers University, New Brunswick, New Jersey, USA. Dr. Faith C. Belanger, Department of Plant Biology, School of Environmental and Biological Sciences, Rutgers University, New Brunswick, New Jersey, USA.

List of Contributors xix

Preface xxiii

Part I Production of Vanilla – Agricultural Systems and Curing 1

1 Mexican Vanilla Production 3
Juan Hernandez‐Hernández

1.1 Introduction 3

1.1.1 The Mexican Vanilla Legend 4

1.2 Cultivation Methods 5

1.2.1 “Traditional”/Acahual 5

1.2.2 Intensive System (Monoculture) 6

1.2.3 Vanilla Cultivation in Existing Orange Groves 6

1.2.4 Shade Houses 7

1.3 Vanilla Propagation Techniques 8

1.3.1 Preparation and Disinfection of Cuttings 8

1.3.2 Establishing Cuttings – Timing 8

1.3.3 Establishing Cuttings – Planting 9

1.3.4 New Bud Formation and Root Growth 9

1.4 Irrigation 9

1.5 Nutrition 10

1.5.1 Mulch 10

1.5.2 Building Compost 10

1.6 Weed Control 11

1.7 Shade Management (Pruning of Support Trees) 11

1.8 Shoot Management – Looping 12

1.9 Shoot Management – Rooting 12

1.10 Main Vanilla Insect Pest 12

1.11 Main Vanilla Diseases 13

1.11.1 Anthracnose 14

1.11.2 Rust 14

1.11.3 Yellowing and Pre‐mature Fruit Drop 14

1.12 Flowering and Pollination 14

1.12.1 Percent of Flowering Plants 15

1.12.2 Natural Pollination 15

1.12.3 Hand Pollination 15

1.12.4 Quantity of Flowers to be Pollinated 17

1.12.5 Fruit Development 17

1.13 Harvesting 17

1.13.1 Harvesting Practices 18

1.13.2 Preventing Theft 18

1.14 Green Vanilla Commercialization 19

1.14.1 Prices 19

1.15 Curing 19

1.15.1 Yield Ratio of Green/Cured Vanilla 21

1.16 Grading 21

1.16.1 Packing 22

1.17 Buyers 23

1.18 Export Volume 23

1.19 Prices 23

1.20 Aromatic Profile 23

1.21 Summary 24

References 24

2 Vanilla Diseases 27
Juan Hernandez‐Hernández

2.1 Introduction 27

2.2 Root and Stem Rot (Fusarium oxysporum f. Sp. Vanillae) 27

2.2.1 Description 27

2.2.2 Damage 28

2.2.3 Control 28

2.3 Black Rot (Phytophtora Sp.) 29

2.3.1 Description 29

2.3.2 Damage 29

2.3.3 Control 29

2.4 Anthracnose (Colletotrichum Sp.) 30

2.4.1 Description 30

2.4.2 Damage 31

2.4.3 Control 31

2.5 Rust (Uromyces Sp.) 31

2.5.1 Description 31

2.5.2 Damage

2.5.3 Control 32

2.6 Rotting of Recently Planted Cuttings 32

2.6.1 Description 32

2.6.2 Damage 32

2.6.3 Control 33

2.7 Yellowing and Shedding of Young Fruits 33

2.7.1 Description 33

2.7.2 Damage 34

2.7.3 Control 34

2.8 Viral Diseases 35

2.8.1 Cymbidium Mosaic Virus (CYMV) 35

2.8.2 Vanilla Mosaic Virus (VMV) 35

2.8.3 Vanilla Necrosis Potyvirus (VNPV) 35

2.8.4 Odontoglossum Ringspot Virus (ORSV) 35

2.8.5 Prevention of Viral Diseases 36

2.9 Damage by Adverse Climatic Factors 36

2.9.1 Natural Pruning of the Apical Buds 36

2.9.1.1 Description 36

2.9.1.2 Damage 37

2.9.1.3 Control 37

2.10 Damage from Sunburn 37

2.10.1 Description 37

2.10.2 Damage 37

2.10.3 Control 38

2.11 Hurricanes 38

References 39

3 Vanilla Production in Costa Rica 41
Elida Varela Quirós

3.1 Introduction 41

3.2 History of Vanilla Production in Costa Rica 42

3.2.1 The First Phase of Large‐scale Cultivation in Costa Rica 42

3.2.2 The Second Phase of Vanilla Cultivation in Costa Rica 42

3.2.3 The Third Phase 43

3.3 Vanilla Production – The Traditional System 45

3.4 Vanilla Production – The Intensive System 47

3.5 Propagation 48

3.6 Diseases and Pests 49

3.7 Vanilla Bean Processing 50

3.8 Conclusions 50

References 51

4 Atypical Flowering of Vanilla planifolia in the Region of Junín, Peru 53
Juan Hernández-Hernández

4.1 Preparation of the “Mother” Plant (Cuttings) 54

4.2 Planting Method 54

4.2.1 Weed Control 55

4.2.2 Shoot Management – Looping 55

4.2.3 Shoot Management – Rooting 55

4.3 Nutrition 55

4.4 Irrigation 55

4.5 Pests, Disorders, and Diseases 57

4.5.1 Vanilla Pest 57

4.5.2 Diseases 57

4.5.3 Intense Solar Radiation 57

4.5.4 New Pest 57

4.5.5 New Disease 58

4.6 Flowering Period 59

4.6.1 Atypical Vanilla Bloom in Peru 59

4.7 Hand Pollination 60

4.8 Harvesting 61

4.9 Vanilla Curing 62

4.10 Final Comments 62

References 63

5 Vanilla Production in the Context of Culture, Economics, and Ecology of Belize 65
Nelle Gretzinger and Dawn Dean

5.1 Introduction 65

5.1.1 Toledo Agriculture and Socio‐demographics Today 66

5.1.2 Maya Mountain Research Farm 66

5.1.3 Agro‐ecological Systems 67

5.1.4 Maya Mountain Research Farm Vanilla Cultivation and Introduction Project 68

5.1.5 The Belize Organic Vanilla Association 69

5.1.6 OVA Description and Goals 69

5.1.7 Innovative Vanilla Plantation Establishment Method Pioneered by OVA Members Nicasio and Ophelia Chee Sanchez 71

5.1.8 Wild/Relic Vanilla Stands in Toledo District 72

5.1.9 Possibility of Wild Superior or Useful Genotypes/Species 74

5.1.10 Dr Pesach Lubinsky’s Research in Belize and Regarding Vanilla tahitensis 74

5.1.11 Manche Chol 76

5.2 Discussion 78

Acknowledgments 79

References 82

6 Conservation and Sustainable Use of Vanilla Crop Wild Relatives in Colombia 85
Nicola S. Flanagan, Paul Chavarriaga, and Ana Teresa Mosquera‐Espinosa

6.1 Introduction 85

6.1.1 Low Genetic Diversity in the Vanilla Crop 85

6.1.2 The Importance of Crop Wild Relatives for Agriculture 85

6.2 Vanilla Crop Wild Relatives 86

6.2.1 Phylogenetic Diversity Within the Genus Vanilla 86

6.2.2 The Secondary Gene Pool for Vanilla 86

6.2.3 Vanilla Diversity in Colombia 87

6.3 Vanilla Species in the Wild 89

6.3.1 Vanilla Species are Rare in the Wild 89

6.3.2 Reproductive Biology of Vanilla Wild Species 91

6.3.2.1 Pollinators 91

6.3.2.2 Autogamy 91

6.3.3 Mycorrhizal Interactions 92

6.3.4 Further Interactions with the Microbiome 93

6.3.5 Bioclimatic and Biophysical Adaptations 94

6.4 Conservation of Vanilla Crop Wild Relatives 95

6.4.1 Threats to Conservation 95

6.4.2 Conservation In situ 96

6.4.3 Conservation Ex situ 96

6.4.4 Conservation Ex situ of the Vanilla Microbiome 98

6.4.5 Conservation of Circa situm and Sustainable Use 98

6.5 Biotechnological Approaches for Vanilla Genetic Resource Conservation and Utilization 100

6.5.1 Characterization and Utilization of Genetic Diversity 100

6.5.1.1 DNA Barcoding 100

6.5.1.2 Genomic Characterization of Vanilla 100

6.5.2 Application of Microorganisms in Vanilla Cultivation 101

6.6 An Integrated Strategy for Conservation and Sustainable Use of Vanilla Crop Wild Relatives 101

6.6.1 A Colombian National Strategy for Vanilla CWR 101

6.6.2 International Strategy for Conservation of Vanilla CWR 102

References 102

7 The History of Vanilla in Puerto Rico: Diversity, Rise, Fall, and Future Prospects 111
Paul Bayman

7.1 Introduction 111

7.2 Diversity of Wild Vanilla in Puerto Rico 111

7.2.1 Species and Distributions 111

7.2.2 Flowering, Pollination, and Fruit Set 112

7.3 Rise and Fall: The History of Vanilla Cultivation in Puerto Rico 112

7.4 Socioeconomic Factors Contributing to the Decline of Vanilla 114

7.5 Diseases and Decline 114

7.5.1 Fusarium Root and Stem Rot (RSR) 115

7.5.1.1 The Pathogen 115

7.5.1.2 Symptoms of RSR 116

7.5.1.3 Other Fusarium Species 116

7.5.2 Other Diseases and Pests 116

7.5.3 Possible Solutions to RSR 116

7.5.3.1 Biological Control 116

7.5.3.2 Mycorrhiza 117

7.5.3.3 Chemical Control 117

7.5.3.4 Breeding 117

7.5.3.5 Cultural Control 117

7.6 Future Prospects 118

Acknowledgments 118

References 118

8 Origins and Patterns of Vanilla Cultivation in Tropical America (1500–1900): No Support for an Independent Domestication of Vanilla in South America 121
Pesach Lubinsky, Gustavo A. Romero‐González, Sylvia M. Heredia, and Stephanie Zabel

8.1 Introduction 121

8.1.1 I. Pre‐Cultivation, ca. 1500–1750s 127

8.1.2 II. Papantla Monopoly, 1760s–1840s 131

8.1.3 III. The Vanilla Revolution, 1850s–1900, “… and we’ve never looked back” 135

8.2 The Vanilla Necklace 136

8.3 Summary 138

Acknowledgments 139

References 139

9 Vanilla Production in Australia 147
Richard Exley

9.1 Introduction 147

9.2 History 147

9.3 Species 148

9.4 Climatic Regions of Australia Suitable for Vanilla 148

9.5 Climatic Conditions in the Vanilla Growing Regions 149

9.6 Soil and Nutrients 150

9.7 Watering 150

9.8 Fertilizing 150

9.9 Propagation 150

9.10 Support 151

9.11 Light/Shade 152

9.12 Spacing 153

9.13 Training 154

9.14 Flowering, Fruit Set, Growth, and Maturation 154

9.14.1 Flowering 154

9.14.2 Fruit Set (Pollination) 154

9.14.3 Growth and Maturation 155

9.15 Harvesting 155

9.16 Curing 155

9.16.1 Overview 155

References 156

10 Vanilla in Dutch Greenhouses: A Discovery – From Research to Production 157
Filip van Noort

10.1 Introduction 157

10.1.1 Start of Research 157

10.2 Review of Literature 157

10.3 Flowering 159

10.3.1 Greenhouse 160

10.3.2 Sustainability 160

10.4 Varieties 161

10.5 Propagation 161

10.5.1 Cultivation 161

10.5.2 Growing Systems 162

10.6 Feasibility and Conclusions 162

References 163

11 Establishing Vanilla Production and a Vanilla Breeding Program in the Southern United States 165
Alan H. Chambers

11.1 Introduction 165

11.2 Southern Florida Climate 165

11.2.1 Average Temperatures 166

11.2.2 Average Rainfall 166

11.2.3 Average Solar Radiation 166

11.2.4 Major Weather Events 168

11.3 Native and Naturalized Vanilla Species of South Florida 169

11.3.1 V. dilloniana 169

11.3.2 V. mexicana 169

11.3.3 V. barbellata 169

11.3.4 V. phaeantha 169

11.3.5 V. planifolia 171

11.4 Establishing Vanilla Production in Southern Florida 173

11.4.1 Shade House Cultivation 173

11.4.2 Tutor Tree Cultivation 173

11.4.3 Substrate Considerations 174

11.4.4 Local Economics and Niche Opportunities 174

11.5 Vanilla Breeding 175

11.5.1 Establishing a Vanilla Breeding Program in the United States 175

11.5.2 Acquiring Diverse Vanilla Accessions 176

11.5.3 Creating Diversity in Vanilla 176

11.5.4 Identifying the Primary Gene Pool 177

11.5.5 Target Traits 177

11.5.6 A Case for a Publically Available Vanilla Genome 178

11.6 Conclusions 178

References 178

12 In vitro Propagation of Vanilla 181
Rebeca Alicia Menchaca García

12.1 Methods 182

12.1.1 In vitro Germination 182

12.1.2 Tissue Culture 182

12.2 Results and Discussion 183

12.2.1 Germination 183

12.2.2 Seed Maturity 183

12.2.3 Time for Germination 183

12.2.4 Scarification 183

12.2.5 Tissue Culture 183

12.2.6 Hybridization 184

12.2.7 In vitro Germplasm Bank 185

12.2.8 Repatriation and Recovery of Mexican Species 185

12.2.9 Method of Ex vitro Adaptation 186

12.2.10 Greenhouse Collection 186

12.2.11 Social Linkage 186

12.2.12 Human Resource Training and International Interaction 187

12.3 Conclusions 187

References 188

13 Curing of Vanilla 191
Chaim Frenkel, Arvind S. Ranadive, Javier Tochihuitl Vázquez, and Daphna Havkin‐Frenkel

13.1 Introduction 191

13.2 Botany of the Vanilla Pod 192

13.2.1 Two Fruit Regions 192

13.2.2 Fruit Components 192

13.2.3 Fruit Anatomy 193

13.2.4 Pollination Initiates Ovary and Fruit Development 193

13.2.5 Mature Fruit 194

13.3 On‐the‐vine Curing Process in a Vanilla Pod 195

13.4 Off‐the‐vine Curing Process of Vanilla Beans 196

13.4.1 Purpose of Curing 198

13.4.2 Traditional Methods of Curing 199

13.4.2.1 Killing 199

13.4.2.2 Sweating 200

13.4.2.3 Drying and Conditioning 201

13.5 Activity of Hydrolytic Enzymes Occurring in a Curing Vanilla Pod 202

13.5.1 Protease Activity 202

13.5.2 Cell Wall Hydrolyzing Enzymes 204

13.5.3 Glycosyl Hydrolases 204

13.6 Activity of Oxidative Enzymes Occurring in a Curing Vanilla Pod 209

13.7 Vanilla Products 212

13.8 Summary and Conclusions 212

13.9 Addendum: Commercial Curing Methods of Green Vanilla Bean 213

13.9.1 Traditional Methods 213

13.9.1.1 Mexican Curing Method 213

13.9.1.2 The Bourbon Curing Method 214

13.9.1.3 The Tahitian Curing Method 214

13.9.1.4 Other Traditional Curing Methods 214

13.9.1.5 Indonesian Curing of Vanilla Bean 215

13.9.2 Refinement of Traditional Curing Methods 215

13.9.3 Novel Curing Methods 215

References 216

14 Fair Trade – The Future of Vanilla? 223
Richard J. Brownell Jr

14.1 The Crisis 223

14.2 The Farmer 224

14.3 Fast Forward 226

14.4 Fair Trade – Background 226

14.4.1 Fair Trade Principles 227

14.4.2 Vanilla and Fair Trade 228

14.5 Commodity Cycles 229

14.6 Issues 230

14.6.1 The Price Differential 230

14.6.2 Vanilla Quality 231

14.6.3 Limited Availability 231

14.6.4 Ensuring that Farmers are Paid the FT Price 232

14.6.5 Consumer Acceptance 232

14.7 Conclusions 233

14.7.1 Update 2017 – Fair Trade Vanilla: Today 233

14.7.2 Update 2017 – Fair Trade Vanilla: The Future 234

Part II Authentication and Flavor Analysis 237

15 Quality Control of Vanilla Beans and Extracts 239
Arvind S. Ranadive

15.1 Introduction 239

15.2 Quality Control of Vanilla Beans 239

15.2.1 Grading of Vanilla Beans 240

15.2.1.1 Vanilla Grading in Mexico 241

15.2.1.2 Vanilla Grading in Madagascar 241

15.2.1.3 Vanilla Grading in Indonesia 241

15.2.1.4 Vanilla Grading in Uganda 241

15.2.1.5 Vanilla Grading in Tahiti 242

15.2.2 Aroma of Vanilla Beans 243

15.2.3 Moisture Content of Vanilla Beans 246

15.2.4 Vanillin Content 246

15.2.4.1 Vanilla Bean Extraction 247

15.2.4.2 Vanillin Determination 247

15.2.4.3 Vanillin Determination in Vanilla Extracts and Other Vanilla Products 248

15.2.4.4 HPLC Method 248

15.2.5 Microbial Contaminant Limits 249

15.3 Quality Control of Commercial Vanilla Products 249

15.3.1 Definition of Vanilla Products 249

15.3.1.1 Vanilla Extracts 249

15.3.1.2 Vanilla Flavoring 250

15.3.1.3 Vanilla‐Vanillin Extract and Flavoring 250

15.3.1.4 Concentrated Vanilla Extract and Flavoring 250

15.3.1.5 Vanilla Oleoresin 250

15.3.1.6 Vanilla Absolute 250

15.3.1.7 Vanilla Powder And Vanilla‐Vanillin Powder 251

15.3.1.8 Vanilla Tincture for Perfumery 251

15.3.2 Vanilla Extract Quality Parameters 251

15.3.2.1 Appearance: Color and Clarity 251

15.3.2.2 Flavor 251

15.3.2.3 Soluble Solids Content 252

15.3.2.4 Vanillin Content 252

15.3.2.5 Organic Acids – (Wichmann) Lead Number 253

15.3.2.6 Resin Content 253

15.3.2.7 Microbial Limits 253

15.4 Determination of Authenticity of Vanilla Extracts 254

15.4.1 Guidelines for Determination of Authenticity 254

15.4.1.1 Evaluation of the Ratios Between Specific Components 255

15.4.1.2 Isotope‐ratios Mass Spectrometry 255

15.4.1.3 Site‐specific Quantitative Deuterium NmR 255

15.4.2 Other Methods to Determine Authenticity 256

15.4.2.1 Stable Isotope Ratio Analysis (SIRA) 256

15.4.2.2 SNIF‐NMR Technique 258

15.5 Summary 259

Acknowledgment 259

References 259

16 Flavor, Quality, and Authentication 261
Patrick G. Hoffman and Charles M. Zapf

16.1 Introduction 261

16.2 Vanilla Flavor Analyses 262

16.3 Biochemistry and Genetic Research on Vanilla 266

16.4 Vanilla Quality and Authentication Analyses 267

16.4.1 Liquid Chromatographic Methods 268

16.4.2 Isotopic Techniques 272

16.4.3 Radiometric and Stable Isotope Ratio Analysis 272

16.4.4 Nuclear Magnetic Resonance (NMR) 274

16.4.5 Isotopic Techniques Summary 274

16.4.6 Integrated and Miscellaneous Methodologies 275

16.5 Conclusion 277

References 279

17 Volatile Compounds in Vanilla 285
Stephen Toth, Keun Joong Lee, Daphna Havkin‐Frenkel, Faith C. Belanger, and Thomas G. Hartman

17.1 Lexicon of Vanilla Aroma/Flavor Descriptors 285

References 345

18 A Comprehensive Study of Composition and Evaluation of Vanilla Extracts in US Retail Stores 349
Daphna Havkin‐Frenkel, Faith C. Belanger, Debra Y.J. Booth, Kathryn E. Galasso, Francis P. Tangel, and Carlos Javier Hernández Gayosso

18.1 History 349

18.2 Uses of Vanilla in the Industry 349

18.2.1 Household Products 350

18.2.2 Dairy Products 350

18.2.3 Ice Cream (Frozen Dairy Products) 350

18.2.4 Yogurt 350

18.2.5 Puddings 351

18.2.6 Chocolate 351

18.2.7 Confections 351

18.2.8 Baked Goods 351

18.2.9 Beverages 351

18.2.10 Pet Products 352

18.2.11 Pharmaceutical Products 352

18.2.12 Oral Care 352

18.2.13 Perfume 352

18.2.14 Toys 352

18.3 Major US Vanilla Companies 353

18.4 Introduction to the Study 353

18.5 Materials and Methods 353

18.6 Results and Discussion 354

18.6.1 Labeling of Retail Vanilla Extracts 354

18.6.2 Flavor Components in the Retail Vanilla Extracts 359

18.6.3 Total Phenol Content of the Retail Vanilla Extracts 363

18.7 Conclusion and Recommendation 363

References 365

19 Vanilla in Perfumery and Beverage Flavors 367
Felix Buccellato

19.1 Earliest Recorded Use of Vanilla 367

Reference 373

Part III Biology of Vanilla 375

20 Vanilla Phylogeny and Classification 377
Kenneth M. Cameron

20.1 Vanilloideae Among Orchids 381

20.2 Diversity Within Vanilloideae 381

20.2.1 Tribe Pogonieae 382

20.2.2 Tribe Vanilleae 383

20.3 Origins and Age of Vanilloideae 384

20.4 Diversity Within Vanilla 385

20.5 Systematic Conclusions and Implications 388

References 389

21 Molecular Analysis of a Vanilla Hybrid Cultivated in Costa Rica 391
Faith C. Belanger and Daphna Havkin‐Frenkel

21.1 Methods 392

21.1.1 PCR Amplification, Cloning, and DNA Sequencing 392

21.1.2 Phylogenetic Analysis 393

21.1.3 Preparation of Vanilla Extracts 393

21.2 Results and Discussion 393

References 399

22 Root Cause: Mycorrhizal Fungi of Vanilla and Prospects for Biological Control of Root Rots 403
Paul Bayman, María del Carmen A. Gonzalez‐Chávez, Ana T. Mosquera‐Espinosa, and Andrea Porras‐Alfaro

22.1 Introduction 403

22.1.1 Orchids and Their Mycorrhiza 403

22.1.2 The Fungi: Rhizoctonia and Related Taxa 404

22.2 Phylogenetic Diversity of Mycorrhizal Fungi of Vanilla 406

22.2.1 Methods 406

22.2.2 Diversity of Mycorrhizal Fungi 408

22.2.3 Fusarium 409

22.2.4 Distribution of Mycorrhiza and Colonization of Roots 409

22.2.5 Roots in Soil vs. Roots on Bark 410

22.2.6 Differences in Mycorrhiza Among Agrosystems 410

22.2.7 Limitations of Methods and Sources of Bias 410

22.3 Mycorrhizal Fungi of Vanilla Stimulate Seed Germination and Seedling Growth 411

22.3.1 Seedling Germination Experiments 411

22.3.2 Seedling Growth and Survival Experiments 411

22.4 Can Mycorrhizal Fungi Protect Vanilla Plants from Pathogens? 414

22.4.1 Biocontrol of Plant Diseases Using Arbuscular Mycorrhizal Fungi 414

22.4.2 Biocontrol of Plant Diseases Using Ceratobasidium 415

22.4.3 Are Rhizoctonia Strains Used for Biocontrol also Potential Pathogens? 416

22.4.4 Fusarium Species as Potential Biocontrol Agents to Protect Vanilla from Fusarium oxysporum Root Rots 417

22.5 Conclusions 417

References 418

23 Enzymes Characterized From Vanilla 423
Andrzej Podstolski

23.1 L‐Phenylalanine Ammonia‐Lyse (Pal) and Cinnamate‐4‐Hydroxylase (C4h) 423

23.2 Chain-shortening Enzymes 424

23.3 4‐Coumaric Acid 3‐Hydroxylase (C3H) 427

23.4 O‐Methyltransferase (OMT) 428

23.5 Benzyl Alcohol Dehydrogenase (Bad) 428

23.6 Glycosyltransferases (GTS) 429

23.7 β‐Glycosyl Hydrolases and Curing 430

References 431

24 Vanillin Biosynthesis – Still not as Simple as it Seems? 435
Richard A. Dixon

24.1 Introduction 435

24.2 Multiple Pathways to Vanillin Based on Biochemistry? 438

24.3 Elucidation of Vanillin Biosynthesis via Molecular Biology? 440

References 442

25 Vanilla planifolia – The Source of the Unexpected Discovery of a New Lignin 447
Fang Chen and Richard A. Dixon

25.1 Introduction 447

25.2 Identification of C‐lignin in V. planifolia 449

25.3 Identification of Genes Potentially Involved in Lignin and Vanillin Biosynthesis 451

25.4 C‐Lignin Biosynthesis in Other Plants 452

25.5 Commercial Value of C‐Lignin as a Novel Natural Polymer 453

References 454

Part IV Biotechnological Production of Vanillin 457

26 Biotechnology of Vanillin: Vanillin from Microbial Sources 459
Ivica Labuda

26.1 Introduction 459

26.1.1 Why? 459

26.1.2 How? 460

26.2 Substrates 460

26.2.1 Ferulic Acid (4‐Hydroxy 3‐Methoxy Cinnamic Acid) 460

26.2.1.1 Non‐β‐oxidative Deacetylation (CoA‐dependent) 462

26.2.1.2 β‐Oxidative Deacetylation (CoA‐Dependent) 463

26.2.1.3 Non‐Oxidative Decarboxylation 464

26.2.1.4 CoA‐Independent Deacetylation 465

26.2.1.5 Side‐Chain Reductive Pathway 466

26.2.2 Eugenol and Isoeugenol 467

26.2.3 Lignin 468

26.2.4 Sugars 469

26.3 Microorganisms 470

26.3.1 Bacteria 470

26.3.1.1 Pseudomonas 470

26.3.1.2 Streptomyces 470

26.3.1.3 Bacillus 471

26.3.1.4 Corynebacterium 472

26.3.1.5 Escherichia coli 472

26.3.1.6 Amycolatopsis sp. 473

26.3.1.7 Lactic Acid Bacteria (LAB) 473

26.3.1.8 Clostridium 474

26.3.2 Fungi and Yeasts 474

26.4 Processes 477

26.4.1 Direct Bioconversion Process 477

26.4.2 Bi‐Phasic Fermentation 480

26.4.3 Mixed Culture Fermentation 480

26.4.4 Continuous Fermentation with Immobilized Cells 481

26.4.5 Enzymes 481

26.4.6 Cofactors 482

26.5 Downstream Processing and Recovery 482

26.6 Conclusions 482

References 483

Index 489