Biosurfactants (eBook)

Ramkrishna Sen is an Assistant Professor (Bioprocess Engineering) in the Department of Biotechnology, Indian Institute of Technology (IIT) Kharagpur, India. Before joining IIT Kharagpur, Dr. Sen served BITS, Pilani as an assistant professor and Cadila Pharmaceuticals Ltd., Ahmedabad as the Manager (R&D–Biotech). He successfully completed some industrial projects and launched modern biotechnology products. In IIT Kharagpur, he currently heads the ‘Bioprocess and Bioproduct Development’ group, consisting of 11 research scholars, 2 MTech and 2 BTech project students, who are actively involved in developing, optimizing, modeling and scaling up bioprocesses for the production and applications of marine biosurfactants, probiotics based nutraceuticals, water-repellant durable jute geotextiles and biofuels. Dr. Sen, being a biochemical engineer with industrial R&D experience had set his research priorities in broader areas of biotherapy and bioenergy. His Biosurfactant group is engaged in characterizing marine microbial surfactants for their potential commercial, healthcare (antimicrobial and anticancer) and environmental (including bioremediation and MEOR) applications. His Nutraceutical group is developing probiotic based nutraceuticals and also working on probioactive molecules like bacteriocin and antihyperglycemic EPS molecule with significant antioxidant activities (Patent Application No.: 594/KOL/2009) and industrial enzymes. Dr. Sen was also involved in developing a biofuel additive for diesel engine, which showed superior fuel properties and pollution characteristics (Patent Appl. No.: KOL /1373/2006). His group is recently involved in developing hydrophobic geotextiles, continuous processes for biodiesel production (sponsored by PfP Technology LLC., Houston, USA) and process integration for bio-ethanol (in collaboration with NEERI, Nagpur) production. Dr. Sen has international research collaborations with some foreign universities and has visited many foreign countries including USA, UK , Brazil, Portugal, Czech Republic, Malaysia, Australia, etc. He has a number of sponsored research and consultancy projects and several research/review articles and book chapters in high impact international journals and highly rated books in the field of biotechnology and biochemical engineering. He serves as a reviewer of 17 peer reviewed international journals and has edited this book being published by Landes Biosciences and Springer Science+Business Media, LLC. Dr. Sen was recently invited as one of the founding members of the recently launched Global Biorenewables (BioEnergy) Research Society (GBR Society) in Lisbon. His biography has been published in Who’s Who in Science & Engineering (2007) and Who’s Who in the World (2008).

Title Page 3
Copyright Page 4
DEDICATION 5
FOREWORD 6
PREFACE 9
ABOUT THE EDITOR... 11
PARTICIPANTS 12
Table of Contents 18
Chapter 1 Screening Concepts for the Isolation of Biosurfactant Producing Microorganisms 26
Introduction 26
Sampling 27
Isolation 27
Screening Methods 28
Surface/Interfacial Activity 28
Direct Surface/Interfacial Tension Measurements 28
Du-Nouy-Ring Method 29
Stalagmometric Method 29
Pendant Drop Shape Technique 29
Axisymmetric Drop Shape Analysis by Profile 30
Measurements Based on Surface/Interfacial Tension 30
Drop Collapse Assay 31
Microplate Assay 31
Penetration Assay 31
Oil Spreading Assay 32
Emulsification Capacity Assay 32
Solubilization of Crystalline Anthracene 33
Cell Surface Hydrophobicity 33
Bacterial Adhesion to Hydrocarbons Assay (BATH) 33
Hydrophobic Interaction Chromatography (HIC) 33
Replica Plate Assay 34
Salt Aggregation Assay 34
Specialities 34
CTAB Agar Plate 34
Hemolysis 35
High Throughput Screening 36
Conclusion and Perspectives 36
References 37
Chapter 2 Molecular Genetics of Biosurfactant Synthesis in Microorganisms 39
Introduction 39
Important Aspects Pertaining to Biosurfactant Production in Microorganisms 40
Molecular Genetics of Biosurfactant Production in Bacteria 40
Acinetobacter Species 40
Emulsan 41
Apoemulsan 42
Alasan 44
Biodispersan 44
Exopolysaccharide (EPS) 44
Pseudomonas Species 44
Bacillus Species 48
Serratia Species 50
Molecular Genetics of Glycolipid Synthesis in Fungi and Yeast 54
Candida 54
Mycobacterium, Corynebacteria, Rhodococcus 55
Pseudozyma, Ustilago maydis 55
Exploitation of Biosurfactant Molecular Genetics in Biotechnological Applications 56
Conclusion 58
Future Prospects 58
Acknowledgements 58
References 59
Chapter 3 Interaction of Dirhamnolipid Biosurfactants with Phospholipid Membranes: A Molecular Level Study 67
Introduction 67
Critical Micellar Concentration of diRL 68
Partitioning of DiRL into Phospholipid Membranes 70
Effect of Membrane Lipid Composition on Membrane Partitioning 70
Modulation of the Thermotropic Behavior of Phospholipids by diRL 71
Effect of diRL on Phospholipid Polymorphism 74
DiRL Affects Phospholipid Acyl Chain Mobility 75
Conclusion 76
References 77
Chapter 4 Microbial Surfactants and Their Potential Applications:An Overview 79
Introduction 79
Classification of Biosurfactants 79
Glycolipids 79
Trehalose Lipids 80
Rhamnolipids 80
Sophorolipids 80
Mannosylerythritol Lipids 80
Lipopeptides 80
Surfactin 80
Iturin 81
Fengycin 81
Lichenysin 81
Fatty Acid Biosurfactant 81
Polymeric Biosurfactants 81
Emulsan 81
Biodispersan 81
Alasan 82
Liposan 82
Emulsifying Biopolymer from Fungus 82
Emulsifying Protein 82
Particulate Biosurfactant 82
Potential Applications of Biosurfactant 82
Role of Microbial Surfactants in Bioremediation of Oil Pollutants 82
Application of Biosurfactant in Petroleum Industry 83
Biosurfactant in Oil Clean Up of Storage Tanks 83
Microbial Surfactants in Microbial Enhanced Oil Recovery (MEOR) 83
Use of Biosurfactants in Food Industries 84
Use of Biosurfactants in Agricultural Sectors 84
Application of Biosurfactant as a Substitute of Synthetic Chemical Surfactant in Commercial Laundry Detergents 84
Biosurfactant as Biopesticide 84
Use of Biosurfactants in Pharmaceutical Sectors and Molecular Biology Research 85
Conclusion 85
References 85
Chapter 5 Microbial Biosurfactants and Biodegradation 90
Introduction 90
Accession of Hydrophobic Contaminants in Aqueous Media 91
Impact of Micellization on Access 93
Accession of Hydrophobic Contaminants in Soil 94
Physiological and Morphological Changes Due to Surfactant Activity 95
Biofilm Formation and Detachment 96
Conclusion 97
References 97
Chapter 6 Biomedical and Therapeutic Applications of Biosurfactants 100
Introduction 100
Biomedical and Therapeutic Applications of Biosurfactants 101
Biological Activity 101
Glycolipids 101
Lipopeptides 104
Other Biosurfactants 106
Anti-Adhesive Activity 107
Antimicrobial Activity 107
Conclusion 108
References 108
Chapter 7 Microbial Surfactants of Marine Origin: Potentials and Prospects 113
Introduction 113
Marine Biosurfactants and Bioemulsifiers 114
Exopolysaccharide Biosurfactants 114
Glycolipopeptides and Carbohydrate-Lipid-Protein Complexes 117
Glycolipids 117
Lipopeptides 119
Environmental and Industrial Potentials 120
Biological Action of the Marine Biosurfactants 123
Conclusion 125
References 125
Chapter 8 Biomimetic Amphiphiles: Properties and Potential Use 127
Introduction 127
Scope 129
Surfactant Basis 129
Specific Classes of Surfactants 130
Cationics 131
Non-Ionics 131
Zwitterionics 131
Surface Active Compounds are Ample in Nature 132
Self-Assembly Processes 133
Association Colloids 133
Micelles 134
Classical Theories of Micelle Formation 135
Micelles and Ahead 135
Emulsification 135
Biosurfactants and Their Potential Uses 136
Activity of Biosurfactants 136
Classification of Biosurfactants 137
Glycolipids 137
Lipopeptides and Lipoprotiens 138
Fatty Acids, Neutral Lipids, Phospholipids 139
Polymeric Surfactants 139
Particulate Biosurfactants 139
Properties of Biosurfactants 140
Surface and Interfacial Activity 140
Temperature, pH and Ionic Strength Tolerance 140
Biodegradability 140
Emulsion Forming and Emulsion Breaking 140
Chemical Diversity 141
Low Toxicity 141
Potential Applications of Biosurfactants 141
Microbial Enhanced Oil Recovery (MEOR) 141
Hydrocarbon Degradation 141
Hydrocarbon Degradation in the Soil Environment 141
Hydrocarbon Degradation in Aquatic Environment 142
Biosurfactant and HCH Degradation 142
Association Properties of Biosurfactants 142
Toxological and Ecological Aspects of Surfactants 143
Dermatological Aspects 143
Aquatic Toxicity 143
Bioaccumulation 143
Biodegradability 144
Conclusion 144
References 144
Chapter 9 Applications of Biological Surface Active Compounds in Remediation Technologies 146
Microbial Surface Active Compounds 146
Structures and Properties 146
Novel Microbial Surface Active Compounds 147
The Roles of SACs in Hydrocarbon Metabolism 148
Microbial Access to Hydrocarbons 148
Altering Access Mode 149
Remediation Technologies 149
Bioremediation 151
Emulsification 151
Micellarization 151
Regulation of Adhesion-Deadhesion of Microorganisms to Hydrocarbons 151
Desorption of Contaminants 155
Soil Washing 155
Hydrocarbon Contaminated Soils 155
Metal Contaminated Soils 156
Conclusion and Prospects 156
References 156
Chapter 10 Possibilities and Challenges for Biosurfactants Use in Petroleum Industry 160
Introduction 160
Surfactants and Biosurfactants in Petroleum Industry 161
Microbial Enhanced Oil Recovery 161
Injection of Ex Situ Produced Biosurfactants into Oil Reservoirs 162
Injection of Laboratory-Selected Biosurfactant-Producing Microorganisms into Oil Reservoirs 162
Stimulation of Indigenous Biosurfactant-Producing Microorganisms within Oil Reservoirs 163
MEOR Field Trials 163
Crude Oil Transportation in Pipeline 164
Clean-Up of Oil Co U ntainers/Storage Tanks 165
Formulation of Petrochemicals 166
Conclusion and Future Perspectives 168
References 169
Chapter 11 Bacterial Biosurfactants, and Their Role in Microbial Enhanced Oil Recovery (MEOR) 171
Introduction 171
Biosurfactant Producing Bacteria 172
Selection of Biosurfactant Producer 172
Factors Affecting Production of Biosurfactants 173
Factors Affecting Biosurfactant Production 173
Biosurfactant Production by Extremophiles 174
Recovery of Biosurfactant 175
Biosurfactant Production by Biotransformation 176
Improved Strains for Biosurfactant Production 176
Biosurfactants and Microbial Enhance Oil Recovery (MEOR) 177
Types of MEOR 177
The Science of MEOR 177
Conclusion and Future Perspectives 178
References 179
Chapter 12 Molecular Engineering Aspects for the Production of New and Modified Biosurfactants 183
Introduction 183
Lipopetides as Targets for Engineering 184
Common Strategies for the Engineering of Biosurfactants 186
Surfactin and Daptomycin as Case Studies for Applied Lipopetide Engineering 188
Problems and Considerations for Biosurfactant Engineering 190
Future Aspects for Lipopeptide Engineering as Revealed by Recent Structural Details 191
Conclusion 192
References 193
Chapter 13 Rhamnolipid Surfactants: Alternative Substrates, New Strategies 195
Introduction 195
Substrates 196
Integrated Systems 198
Physicochemical Properties 199
Rhamnolipid Solutions 199
Emulsions and Microemulsions 200
Wetting Properties 201
Effect of Electrolytes 201
Effect of pH on Aggregation Morphology 202
Foam Film 203
Applications 203
Biosurfactants and Petroleum 203
BioSurfactants and Heavy Metals 204
Rhamnolipids and Antimicrobial Activity 204
Biosurfactants in Food, Cosmetics and Pharmaceuticals 205
Conclusion 206
References 207
Chapter 14 Selected Microbial Glycolipids: Production, Modification and Characterization 210
Introduction 210
Glycoglycerolipids 210
General Information 210
Glycoglycerolipids from Eukaryotic Cells 211
Glycoglycerolipids from Prokaryotic Cells 212
Glycoglycerolipids from Synthetical Route 212
Selected Glycoglycerolipids from Prokaryotes 213
Molecular Structures 213
Production, Downstream Processing and Analysis 213
Glucosylmannosyl-Glycerolipid fromMicrobacterium Spec. DSM 12583 213
Dimannosyl-Glycerolipid fromMicrococcus Luteus (Hel 12/2) 216
Diglucosyl-Glycerolipid from Bacillus pumilus Strain AAS3 216
Chemo-Enzymatic Modification of Glycoglycerolipids 217
Oligosaccharide Lipids 218
General Information 218
Selected Oligosaccharide Lipids 220
Molecular Structures 220
Production, Downstream Processing and Analysis 220
Oligosaccharide Lipids from Tsukamurella Spec. DSM 44370 220
Pentasaccharide Lipids from Nocardia corynebacteroides SM1 222
Chemo-Enzymatic Modification of Oligosaccharide Lipids from Tsukamurella spec 223
Physico-Chemical and Bioactive Properties 223
Conclusion 225
References 225
Chapter 15 Production of Microbial Biosurfactants by Solid-State Cultivation 228
Introduction 228
Microbial Biosurfactants That It Would Be Interesting to Produce at Large Scale 229
Production of Biosurfactants by Classical Submerged Cultivation Is Problematic 229
Solid-State Cultivation as an Alternative Cultivation Technique with Potential for Biosurfactant Production 230
What Is the State of the Art of Biosurfactant Production in Solid-State Cultivation? 230
What Challenges Do We Face in the Production of Biosurfactants by Solid-State Cultivation? 232
Bioreactor Selection 232
What Will Be the Best Substrate to Use? 233
Downstream Processing 233
Monitoring of the Cultivation Process 234
Conclusion 234
References 234
Chapter 16 Rhamnolipid Biosurfactants: Production andTheir Potential in Environmental Biotechnology 236
Introduction 236
Chemical Structures and Properties of Rhamnolipid Biosurfactants 237
Biosynthesis of Rhamnolipid Biosurfactants 239
Production of Rhamnolipid Biosurfactants 239
Potential Applications of Rhamnolipid Biosurfactants 242
Conclusion and Future Perspectives 243
References 243
Chapter 17 Biosurfactant’s Role in Bioremediation of NAPL and Fermentative Production 247
Introduction 247
Bioremediation 248
What Are Non-Aqueous Phase Liquids (NAPL)? 248
Chemical Surfactants and Bioremediation 249
Biosurfactants and Bioremediation 250
Fermentative Production and Recovery of Biosurfactants 250
Production of Biosurfactants 251
Recovery of Biosurfactants 252
Economical Commercial Production 252
The Use of Cheaper Waste Substrates 253
Development of Efficient Fermentation Processes 254
Media and Process Optimization 254
Recovery Processes 256
Conclusion 256
References 257
Chapter 18 Biosurfactants from Yeasts: Characteristics, Production and Application 261
Introduction 261
Biosurfactant Classification and Characteristics 262
Production Processes 263
The Influence of the Culture Medium Composition 263
Carbon Source 264
Carbon Source from Renewable Resources 265
Nitrogen Source 266
The Environmental Factors Affecting the Production 267
pH 267
Temperature 267
Aeration and Agitation 270
Kinetics and Operation of Biosurfactant Production Process 270
Potential Commercial Applications 271
Conclusion 272
References 272
Chapter 19 Environmentally Friendly Biosurfactants Produced by Yeasts 275
Introduction 275
Culture Conditions for the Production of Biosurfactants 277
Substrates Used for the Production of Candida Biosurfactant 277
Emulsifying Activities, Surface Tension and Critical Micellar Concentration (CMC) of Candida Biosurfactants 277
Biosurfactant: Isolation Methodology and Yields 278
Biochemical Composition and Application of Biosurfactant 279
Trends and Future Challenges of Biosurfactants 280
Computer-Based Tools for Optimization and Cost Reduction 280
Experimental Design and Surface Response Methodology 280
Novel Tools in Biosurfactant Production, Control and Optimization Processes: Application of an Artificial Neural Network 281
Conclusion 281
References 282
Chapter 20 Synthesis of Biosurfactants and Their Advantages to Microorganisms and Mankind 286
Introduction 286
Surfactants and Biosurfactants 287
Significance and Role of Biosurfactants to Microbes 290
Adhesion 290
Emulsification 291
Bioavailability and Desorption 292
Defense Strategy 292
Advantages of Biosurfactants 293
Biodegradability and Controlled Inactivation of Microbial Surfactants 293
Selectivity for Specific Interfaces 293
Surface Modification 293
Diversity of Microbial Surfactants 293
Toxicity 293
Biosurfactants Types and Producing Organisms 293
Applications of Biosurfactants 294
Biosurfactant and Environment 294
Biosurfactants and Medicine 299
Biosurfactants and Miscellaneous Applications 299
Conclusion 300
References 300
Chapter 21 Enrichment and Purification of Lipopeptide Biosurfactants 306
Introduction 306
Properties of Biosurfactants Useful in Separation 306
Separation by Precipitation 307
Liquid Partitioning 307
Direct from Cell Culture 307
Solvent Extraction as a Means of Purification 307
Membrane Filtration 308
Cross Flow Ultrafiltration 308
Liquid Membranes 308
Foam Fractionation 308
Surface Skimming 310
Surface Enrichment 311
Adsorption to Solids 311
High Performance Liquid Chromatography 311
Conclusion 311
References 312
Chapter 22 Production of Surface Active Compounds by Biocatalyst Technology 314
Introduction 314
Biocatalysis 315
Enzymatic Synthesis of Monoglycerides 316
Synthesis of Sugar Esters 318
Synthesis of Fatty Acid Amides 320
Enzymatic Synthesis of Alkyl Glycosides 321
Yeast and Fungal Glucosidases 322
Plant Derived Glucosidases 323
Glucosidases Derived from Bacteria 323
Enzymatic Synthesis of Phospholipids 323
Enzymatic Production of Lysophospholipids 324
Enzymatic Production of Modified or Structured Phospholipids 325
Conclusion 326
References 326
Chapter 23 Structural and Molecular Characteristics of Lichenysin and Its Relationship with Surface Activity 329
Introduction 329
Surfactin 330
Nonribosomal Peptide Synthesis 332
Lichenysin Structure 332
Lichenysin Operon 335
Lichenysin Synthetase 336
Structure-Activity Relationship 337
Conclusion 338
References 338
Chapter 24 Surfactin: Biosynthesis, Genetics and Potential Applications 341
Introduction 341
Biosynthesis of Surfactin 342
Biochemistry and Mechanisms 342
Role of Genetic Regulations in Surfactin Biosynthesis 343
Potential Commercial Applications 343
Health-Care and Bio-Control Applications 343
Environmental Applications 345
Conclusion 345
References 345
Index 349