Handbook of Microalgal Culture: Applied Phycology and Biotechnology

Description

Algae are some of the fastest growing organisms in the world, with up to 90% of their weight made up from carbohydrate, protein and oil. As well as these macromolecules, microalgae are also rich in other high-value compounds, such as vitamins, pigments, and biologically active compounds, All these compounds can be extracted for use by the cosmetics, pharmaceutical, nutraceutical, and food industries, and the algae itself can be used for feeding of livestock, in particular fish, where on-going research is dedicated to increasing the percentage of fish and shellfish feed not derived from fish meal. Microalgae are also applied to wastewater bioremediation and carbon capture from industrial flue gases, and can be used as organic fertilizer. So far, only a few species of microalgae, including cyanobacteria, are under mass cultivation. The potential for expansion is enormous, considering the existing hundreds of thousands of species and subspecies, in which a large gene-pool offers a significant potential for many new producers. Completely revised, updated and expanded, and with the inclusion of new Editor, Qiang Hu of Arizona State University, the second edition of this extremely important book contains 37 chapters. Nineteen of these chapters are written by new authors, introducing many advanced and emerging technologies and applications such as novel photobioreactors, mass cultivation of oil-bearing microalgae for biofuels, exploration of naturally occurring and genetically engineered microalgae as cell factories for high-value chemicals, and techno-economic analysis of microalgal mass culture. This excellent new edition also contains details of the biology and large-scale culture of several economically important and newly-exploited microalgae, including Botryococcus, Chlamydomonas, Nannochloropsis, Nostoc, Chlorella, Spirulina, Haematococcus, and Dunaniella species/strains. Handbook of Microalgal Culture Applied Phycology and Biotechnology Contents List of Contributors Acknowledgments Introduction Part 1 The Microalgal Cell with Reference to Mass Cultures 1 The Microalgal Cell 1.1 INTRODUCTION 1.2 GROSS MORPHOLOGY 1.3 SEXUAL REPRODUCTION 1.4 ULTRASTRUCTURE 1.4.1 Chloroplast 1.4.2 Mitochondrion 1.4.3 Nucleus and mitosis 1.4.4 Golgi body and endoplasmic reticulum 1.4.5 Vacuoles 1.4.6 Flagella and eyespots 1.4.7 Cell walls and coverings 1.5 BIOCHEMICAL ASPECTS 1.5.1 Carbohydrates 1.5.2 Lipids 1.5.3 Proteins 1.6 BIODIVERSITY 1.7 EVOLUTION AND SYSTEMATIC BIOLOGY 1.7.1 Evolutionary origins 1.7.2 Cyanobacteria 1.7.3 Eukaryotic super groups 1.7.4 Glaucophyte algae 1.7.5 Green algae 1.7.6 Red algae 1.7.7 Heterokont algae 1.7.8 Dinoflagellates 1.7.9 Haptophytes 1.7.10 Cryptophytes 1.7.11 Euglenoids 1.7.12 Chlorarachniophytes 1.7.13 Other photosynthetic alga-like organisms 1.8 ECOLOGY ACKNOWLEDGMENT REFERENCES 2 Photosynthesis in Microalgae 2.1 THE PROCESS OF PHOTOSYNTHESIS 2.2 THE NATURE OF LIGHT 2.3 PHOTOSYNTHETIC PIGMENTS 2.4 THE LIGHT REACTIONS OF PHOTOSYNTHESIS 2.4.1 The photosynthetic membranes 2.4.2 Photosynthetic electron transport and phosphorylation 2.4.3 The outer light-harvesting antennae 2.4.4 Photosystem II 2.4.5 Plastoquinone, the cytochrome b6/f complex, and plastocyanin 2.4.6 Photosystem I 2.4.7 ATP synthase/ATPase 2.5 THE DARK REACTIONS OF PHOTOSYNTHESIS 2.5.1 Carbon assimilation 2.5.2 Photorespiration 2.6 LIGHT ACCLIMATION 2.7 SELECTED MONITORING TECHNIQUES USED IN MICROALGAL BIOTECHNOLOGY 2.7.1 Measurement of photosynthetic oxygen evolution 2.7.2 Measurement of photosynthetic carbon fixation 2.7.3 Chlorophyll fluorescence 2.8 THEORETICAL LIMITS OF MICROALGAL PRODUCTIVITY ACKNOWLEDGEMENT REFERENCES 3 Basic Culturing and Analytical Measurement Techniques 3.1 ISOLATION OF MICROALGAE 3.1.1 Selection of sources of microalgae 3.1.2 Enrichment of a culture 3.1.3 Direct isolation 3.1.4 Producing axenic cultures 3.2 SCREENING OF MICROALGAE FOR BIOACTIVE MOLECULES 3.2.1 Direct assays 3.2.2 Indirect assays 3.3 MAINTENANCE AND PRESERVATION OF MICROALGAL STRAINS 3.4 MEASUREMENT OF GROWTH PARAMETERS 3.4.1 Cell count 3.4.2 Optical density method for determination of microalgal biomass 3.4.3 Dry and wet mass 3.4.4 Moisture content and ash content 3.4.5 Chlorophyll determination 3.4.6 Total organic carbon (TOC) measurement 3.4.7 Doubling time, specific growth rate, and output rate 3.4.8 Growth yield 3.4.9 Maintenance energy requirement 3.5 MODES OF CULTURE 3.5.1 Batch culture 3.5.2 Continuous cultures 3.5.3 Immobilized cultures 3.6 ADVANCED BIOCHEMICAL ANALYSIS 3.6.1 Carbohydrates 3.6.2 Proteins 3.6.3 Lipids 3.6.4 Fatty acid composition analysis 3.6.5 Lipid determination using fluorescence spectroscopy and microscopy ACKNOWLEDGMENT REFERENCES 4 Strategies for Bioprospecting Microalgae for Potential Commercial Applications 4.1 INTRODUCTION 4.2 UNIVARIATE APPROACHES TO DESIGN AND IMPLEMENTATION OF STRAIN COLLECTION STRATEGIES 4.2.1 Collection strategies focused on broad collection of all algae in general 4.2.2 Collection strategies focused on specific habitat types 4.2.3 Collection strategies focused on a targeted chemical composition or product 4.2.4 Collection strategies centered on physiological or chemical attributes of the strains 4.3 MULTIVARIATE APPROACHES TO DESIGN AND IMPLEMENTATION OF STRAIN COLLECTION STRATEGIES 4.3.1 Sample collection and enrichment for targeted strain capabilities 4.3.2 Simultaneous collection and screening for two or more targeted capabilities REFERENCES 5 Maintenance of Microalgae in Culture Collections 5.1 INTRODUCTION 5.2 THE DIVERSITY OF MICROALGAE IN CULTURE COLLECTIONS 5.3 THE CONCEPT OF STRAINS VERSUS SPECIES OF MICROALGAE 5.4 MAINTENANCE OF ACTIVELY GROWING CULTURES 5.4.1 Purity of cultures 5.4.2 Quality control and financial considerations 5.4.3 Cryopreservation 5.4.4 Identifying and authenticating strains of microalgae in culture collections 5.4.5 The future of culture collections ACKNOWLEDGMENT REFERENCES 6 Environmental Stress Physiology with Reference to Mass Cultures 6.1 INTRODUCTION 6.2 LIGHT AND PHOTOSYNTHESIS RATE 6.2.1 P versus I curve 6.2.2 Photoacclimation 6.2.3 Photoinhibition 6.2.4 Photoinhibition in outdoor cultures 6.2.5 Some practical considerations 6.3 SALINITY STRESS 6.4 CONCLUDING REMARKS 6.5 SUMMARY ACKNOWLEDGMENT REFERENCES 7 Environmental Effects on Cell Composition 7.1 INTRODUCTION 7.2 ENVIRONMENTAL FACTORS 7.2.1 Light 7.2.2 Temperature 7.3 NUTRITIONAL FACTORS 7.3.1 Nitrogen 7.3.2 Phosphorus 7.3.3 Iron 7.4 SALINITY 7.5 SYNERGISTIC EFFECTS OF COMBINATIONS OF CHEMICAL AND PHYSICAL FACTORS ON CELL COMPOSITION 7.6 BIOTECHNOLOGICAL APPROACHES TO CONTROL CELL COMPOSITION REFERENCES 8 Inorganic Algal Nutrition 8.1 NUTRITIONAL MODES 8.2 NUTRIENT REQUIREMENTS 8.3 CARBON 8.4 NITROGEN 8.5 PHOSPHORUS 8.6 OTHER MACRO- AND MICRONUTRIENTS, CHELATES, AND WATER 8.7 RECIPES FOR ALGAL GROWTH NUTRIENT MEDIA 8.8 UPTAKE OF N AND P 8.8.1 Competition for limiting resources (nutrients) 8.9 NUTRIENT RATIOS 8.10 PHYSICAL FACTORS INFLUENCING NUTRIENT UPTAKE 8.10.1 Bioremediation REFERENCES 9 Commercial Production of Microalgae via Fermentation 9.1 INTRODUCTION 9.2 WHY HETEROTROPHIC PRODUCTION OF ALGAE? 9.3 THE HETEROTROPHIC CAPACITY OF MICROALGAE 9.4 EARLY HISTORY OF THE PRODUCTION OF MICROALGAE IN COMMERCIAL-SCALE FERMENTORS 9.5 COMMERCIAL SUCCESS: CHLORELLA FERMENTATION FOR NUTRITIONAL SUPPLEMENTS 9.6 COMMERCIAL SUCCESS: CRYPTHECODINIUM FERMENTATION FOR DHA 9.7 COMMERCIAL SUCCESS: SCHIZOCHYTRIUM FERMENTATION FOR DHA 9.8 HETEROTROPHIC CHLORELLA: FUTURE DIRECTIONS 9.8.1 Biodiesel from heterotrophic Chlorella 9.8.2 Food ingredients from heterotrophic Chlorella 9.8.3 Renewable chemicals from heterotrophic Chlorella 9.9 CRYPTHECODINIUM: FUTURE DIRECTIONS 9.10 SCHIZOCHYTRIUM: FUTURE DIRECTIONS 9.10.1 Biodiesel from Schizochytrium 9.10.2 New food ingredients from Schizochytrium 9.11 OTHER HETEROTROPHIC MICROALGAE: FUTURE DIRECTIONS REFERENCES 10 Molecular Genetic Manipulation of Microalgae: Principles and Applications 10.1 GENE STRUCTURE AND CONTROL OVER EXPRESSION 10.1.1 Control elements affecting mRNA levels 10.2 SELECTION MARKERS 10.3 TRANSFORMATION METHODS 10.3.1 Comparison of transformation efficiencies 10.4 GENE TARGETING AND KNOCKDOWNS 10.4.1 Insertional mutagenesis 10.4.2 Homologous recombination 10.4.3 RNAi- and antisense RNA expressionmediated gene knockdown 10.5 PROBLEMS IN ALGAL TRANSGENICS 10.5.1 Gene silencing 10.5.2 Codon usage 10.5.3 Introns and other elements 10.6 NUCLEAR VERSUS CHLOROPLAST TRANSFORMATION 10.7 METABOLIC ENGINEERING 10.7.1 Selection of gene targets for metabolic engineering 10.8 MICROALGAE AS PROTEIN EXPRESSION SYSTEMS 10.9 BREEDING, MUTAGENESIS, AND SELECTION 10.10 SUMMARY AND FUTURE DIRECTIONS ACKNOWLEDGMENT REFERENCES Part 2 Mass Cultivation and Processing of Microalgae 11 Biological Principles of Mass Cultivation of Photoautotrophic Microalgae 11.1 LIGHT: THE MAJOR FACTOR IN GROWTH AND PRODUCTIVITY 11.2 CELL CONCENTRATION: A PROMINENT FACTOR OF THE LIGHT REGIME OF CELLS IN THE CULTURE 11.2.1 Areal and population densities 11.2.2 Light penetration depth 11.2.3 Effect of cell density on cellular ultrastructure and composition 11.3 MIXING PHOTOAUTOTROPHIC CULTURES 11.4 LIGHT–DARK CYCLE FREQUENCIES 11.5 THE OPTICAL PATH, A DECISIVE PARAMETER IN GROWTH AND PRODUCTIVITY OF PHOTOAUTOTROPHIC CULTURES 11.6 ULTRAHIGH CELL DENSITY CULTURES 11.6.1 Growth inhibitory substances and conditions 11.6.2 Areal density in relation to the optical path 11.7 REACTION TIMESCALES IN PHOTOSYNTHESIS, IN RELATION TO THE EFFECT OF THE OPTICAL PATH ON CULTURE PRODUCTIVITY 11.7.1 Reaction timescales in photosynthesis 11.7.2 Cell travel times between the lit and dark volumes in the reactor 11.7.3 Long optical paths 11.7.4 Short optical paths 11.7.5 Radiation dependence of the photosynthetic reaction center – turnover time 11.8 THE AVERAGE RADIATION INTENSITY 11.9 EFFECTIVE USE OF SUNLIGHT AND HIGH IRRADIANCE FOR PHOTOSYNTHETIC PRODUCTIVITY 11.9.1 Response to changing irradiance outdoors 11.9.2 Tilting reactor surfaces in adjustment to the solar angle 11.10 PHOTOSYNTHETIC EFFICIENCY IN MASS CULTURES (SEE CHAPTER 2) 11.10.1 Appraising algal productivity and assessment of reactor efficiency (see also Chapter 12) 11.11 MAINTENANCE OF MASS CULTURES 11.11.1 Online monitoring of photosynthetic activity 11.11.2 Measurement of cell growth and culture productivity 11.11.3 Night biomass loss 11.11.4 Maintaining OPD 11.11.5 Preventing nutritional deficiencies 11.11.6 Maintenance of monoalgal cultures and combating contamination REFERENCES 12 Theoretical Analysis of Culture Growth in Flat-Plate Bioreactors: The Essential Role of Timescales 12.1 INTRODUCTION 12.1.1 The issues addressed in this chapter 12.1.2 Conventional versus thin flat-plate bioreactors 12.1.3 Some relevant timescales 12.2 FLAT-PLATE BIOREACTORS – QUALITATIVE EXPECTATIONS 12.2.1 Reactor geometry and random cell motion 12.2.2 Cell random motion and light regime 12.2.3 Volumetric production rate – effects of culture density and cell random motion 12.2.4 OP length and productivity 12.2.5 Extreme culture densities 12.2.6 Increasing light intensity and photoproductivity 12.3 MODELS FOR PHOTOSYNTHESIS BY RANDOMLY MOVING CELLS 12.3.1 Model I – sequential collection reaction 12.3.2 Model II – ensemble-averaged kinetics 12.3.3 One-step versus four-step model 12.3.4 Parameter values for Model II 12.4 THE IMPORTANCE OF TIMESCALES 12.4.1 Productivity under a flashing light regime 12.4.2 Implications for reactor performance 12.5 RESULTS OF MODEL CALCULATIONS 12.5.1 Extremely high sensitivity to parameter values 12.5.2 No obvious signs of significant photoinhibition as light intensity is raised 12.5.3 Irradiation on one side versus two sides 12.5.4 Results of Model I 12.6 OPEN QUESTIONS 12.6.1 Physical aspects 12.6.2 Dynamical aspects of photosynthesis REFERENCES 13 Photobioreactors for Mass Production of Microalgae 13.1 INTRODUCTION 13.2 OPEN SYSTEMS 13.3 PHOTOBIOREACTORS 13.3.1 Photobioreactors: definition and classification 13.3.2 Vertical columns and sleeves 13.3.3 Tubular photobioreactors 13.3.4 Vertical and inclined flat photobioreactors 13.3.5 Flexible film panels 13.3.6 Hybrid systems 13.3.7 Floating systems 13.3.8 Photobioreactors for attached microalgae 13.3.9 Photobioreactors for special applications 13.3.10 Innovations in illumination devices 13.4 COMBINED PRODUCTION PROCESSES: COUPLING PONDS AND PHOTOBIOREACTORS 13.5 PRODUCTION CHARACTERISTICS AND COST OF RECENTLY DEVELOPED CULTIVATION SYSTEMS 13.6 CONSIDERATIONS ON PRODUCTIVITY AND PHOTOSYNTHETIC EFFICIENCY EVALUATION OF OUTDOOR MICROALGAL CULTURES 13.7 CONCLUSIONS REFERENCES WEB REFERENCES 14 Downstream Processing of Cell Mass and Products 14.1 INTRODUCTION 14.2 MICROALGAE HARVESTING 14.2.1 Coagulation–flocculation 14.2.2 Gravitational separation: sedimentation and flotation 14.2.3 Filtration 14.2.4 Centrifugal recovery 14.2.5 New technologies for harvesting 14.2.6 Criteria for selecting the harvesting method 14.3 DEHYDRATING 14.3.1 Spray drying 14.3.2 Sun drying 14.3.3 Solar drying 14.3.4 Convective hot air drying 14.3.5 Lyophilization 14.4 CELL DISRUPTION 14.4.1 Cell disruption by chemical methods 14.4.2 Cell disruption by mechanical methods 14.5 PRODUCT ISOLATION 14.5.1 Extraction procedures 14.5.2 Expanded bed adsorption 14.6 PRODUCT PURIFICATION 14.7 COST-EFFECTIVENESS: CASE-STUDY ANALYSIS OF MICROALGAL BIOFUEL 14.7.1 The microalgal-to-biodiesel production process 14.7.2 Direct transesterification of wet biomass and recovery of FAMEs 14.7.3 Economic assessment of the entire microalgal-to-biodiesel process 14.8 CONCLUDING REMARKS REFERENCES 15 First Principles of Techno-Economic Analysis of Algal Mass Culture 15.1 BRIEF INTRODUCTION OF KEY CONCEPTS 15.2 PRINCIPLES OF TECHNO-ECONOMIC ANALYSIS 15.3 CONSTRUCTING THE SPREADSHEET TEMPLATE 15.3.1 Model assumptions and unit conversion methods 15.3.2 Key techno-economic analysis output metrics 15.3.3 Data sources 15.3.4 Model inputs: translating a process flow diagram into a techno-economic analysis 15.3.6 Completed TEA 15.4 CONCLUSION REFERENCES Part 3 Commercial Species of Industrial Production 16 Chlorella: Industrial Production of Cell Mass and Chemicals 16.1 INTRODUCTION 16.2 MORPHOLOGY, ULTRASTRUCTURE, AND TAXONOMY 16.3 PHOTOSYNTHESIS, NUTRIENT REQUIREMENTS, AND GROWTH PHYSIOLOGY 16.4 BIOCHEMICAL COMPOSITION 16.5 MASS CULTIVATION 16.5.1 Open culture systems 16.5.2 Closed PBRs 16.5.3 Fermentor 16.6 CHLORELLA HARVESTING AND DRYING 16.7 POTENTIAL PRODUCTS AND APPLICATIONS 16.7.1 Human food and animal feed 16.7.2 Source of carotenoids 16.7.3 Biofuels explorations 16.7.4 CO2 biomitigation 16.7.5 Bioremediation of wastewater 16.7.6 Recombinant proteins 16.8 FUTURE PERSPECTIVE REFERENCES 17 Biology and Industrial Production of Arthrospira (Spirulina) 17.1 TAXONOMY 17.2 MORPHOLOGY AND ULTRASTRUCTURE 17.2.1 Morphology 17.2.2 Ultrastructure 17.3 ECOLOGY 17.4 PHYSIOLOGY 17.4.1 The effect of light on photosynthesis and growth 17.4.2 The effect of temperature on photosynthesis and growth 17.4.3 Salt stress and osmoregulation 17.4.4 Effect of oxygen concentration on photosynthesis, photorespiration, and growth 17.4.5 Nutrient physiology 17.4.6 Heterotrophy and mixotrophy in Arthrospira 17.5 INDUSTRIAL PRODUCTION OF ARTHROSPIRA 17.5.1 History and current production of Arthrospira 17.5.2 The environmental setting for outdoor biomass production 17.5.3 The production process 17.5.4 Harvesting the biomass 17.5.5 Drying and packaging the biomass 17.5.6 Product quality and safety 17.5.7 Regulatory status of Spirulina 17.5.8 Current status and future prospects for industrial production of Spirulina ACKNOWLEDGMENT REFERENCES 18 Dunaliella: Biology, Production, and Markets 18.1 INTRODUCTION 18.2 THE ALGA 18.3 COMMERCIAL-SCALE PRODUCTION 18.4 BIOACTIVITY OF DUNALIELLA AND ITS CONSTITUENTS 18.5 PRODUCERS AND MARKETS REFERENCES 19 Biology and Industrial Potential of Botryococcus braunii 19.1 INTRODUCTION 19.2 PHYLOGENY AND TAXONOMY 19.3 MECHANISM FOR BIOSYNTHESIS OF HYDROCARBONS PRODUCED BY RACES A AND B 19.3.1 Hydrocarbons synthesized by race A 19.3.2 Hydrocarbons synthesized by race B 19.4 CULTURE MEDIA 19.5 EFFECTS OF CULTURE CONDITIONS ON GROWTH AND HYDROCARBON PRODUCTION: TEMPERATURE, SALINITY, PH, CO2 CONCENTRATION, AND NUTRIENTS (N, P, ETC.) 19.5.1 Temperature 19.5.2 Light 19.5.3 Salinity 19.5.4 pH and CO2 concentration 19.5.5 Nutrients (N, P, etc.) 19.6 MIXOTROPHY 19.7 UTILIZATION OF WASTEWATER FOR BOTRYOCOCCUS CULTIVATION 19.8 OUTDOOR CULTIVATION 19.9 BUSINESS EVALUATION ACKNOWLEDGMENT REFERENCES 20 Biology and Commercial Aspects of Haematococcus pluvialis 20.1 INTRODUCTION 20.2 CELL BIOLOGY OF H. PLUVIALIS 20.2.1 Morphology and cell cycle 20.2.2 Ultrastructural changes during carotenogenesis 20.3 CHANGES IN PIGMENT COMPOSITION AND PHOTOSYNTHESIS UNDER OXIDATIVE STRESS 20.4 BIOSYNTHESIS OF ASTAXANTHIN IN H. PLUVIALIS 20.4.1 Enzymes and pathways for astaxanthin biosynthesis 20.4.2 Regulation of carotenogenesis 20.4.3 Crosstalk between astaxanthin and photosynthetic electron transfer chain 20.4.4 Crosstalk between astaxanthin and fatty acid biosynthesis 20.4.5 Genetic improvement of H. pluvialis for enhanced astaxanthin production 20.5 PHYSIOLOGICAL ROLE OF ASTAXANTHIN BIOSYNTHESIS 20.6 EFFECTS OF ENVIRONMENTAL FACTORS ON GROWTH AND PRODUCTIVITY 20.7 MASS CULTIVATION OF H. PLUVIALIS 20.7.1 Photoautotrophic culture 20.7.2 Heterotrophic and mixotrophic culture modes 20.7.3 Microbial contamination and crop protection 20.8 DOWNSTREAM PROCESSES AND PRODUCT FORMATION 20.9 CONCLUSION AND PERSPECTIVES REFERENCES 21 Novel Sulfated Polysaccharides of Red Microalgae: Basics and Applications 21.1 PRODUCTS OF RED MICROALGAE: MAINLY SULFATED POLYSACCHARIDES 21.2 SULFATED POLYSACCHARIDE CHARACTERISTICS AND FUNCTION 21.3 SULFATED POLYSACCHARIDE FORMATION: ENVIRONMENTAL, BIOCHEMICAL, AND GENETIC ASPECTS 21.3.1 Effects of the ambient environment on polysaccharide production 21.3.2 Biochemical pathways 21.3.3 Genetics and genetic engineering 21.4 MASS MICROALGAL CULTIVATION 21.5 APPLICATIONS OF SULFATED POLYSACCHARIDES OF RED MICROALGAE 21.6 THE FUTURE REFERENCES 22 Hydrogen Production by Chlamydomonas reinhardtii 22.1 INTRODUCTION 22.2 ALGAE AS A PLATFORM FOR PRODUCING HYDROGEN 22.2.1 Challenges for developing an algal biohydrogen system 22.3 ALGAL METABOLIC PROCESSES INVOLVED IN HYDROGEN SYNTHESIS 22.3.1 Photosynthesis 22.3.2 Hydrogen-evolving enzymes 22.4 THEORETICAL LIMIT TO PHOTOBIOLOGICAL HYDROGEN PRODUCTION 22.4.1 Scientific barriers to reaching the theoretical efficiency limit 22.4.2 Technical barriers to reaching the theoretical efficiency limit 22.5 BIOTECHNOLOGY OF HYDROGEN PRODUCTION 22.5.1 Hydrogen production in laboratory-scale photobioreactors 22.5.2 Hydrogen production in outdoor photobioreactors 22.5.3 Lessons learned 22.6 MATERIALS FOR HYDROGEN PHOTOBIOREACTOR CONSTRUCTION 22.7 NET ENERGY RATIO FOR PHOTOBIOLOGICAL HYDROGEN PRODUCTION 22.8 CONTAMINATION 22.9 FUTURE PROSPECTS FOR COMMERCIAL APPLICATIONS ACKNOWLEDGMENT REFERENCES 23 Biology and Biotechnology of Edible Nostoc 23.1 INTRODUCTION 23.2 MORPHOLOGICAL FEATURES AND COLONIAL DEVELOPMENT IN EDIBLE STRAINS 23.3 SPECIES IDENTIFICATION BY USING MOLECULAR MARKERS 23.4 NUTRITIONAL AND PHARMACEUTICAL VALUE OF NOSTOC 23.4.1 Nutritional value 23.4.2 Bioactivities of Nostoc polysaccharides 23.4.3 Other Nostoc bioactive compounds 23.5 GROWTH AND PHYSIOLOGY 23.6 MASS CULTIVATION REFERENCES 24 IGV GmbH Experience Report, Industrial Production of Microalgae Under Controlled Conditions: Innovative Prospects 24.1 INTRODUCTION 24.2 CONTROLLING CLOSED SYSTEMS FOR MICROALGAE PRODUCTION 24.3 PBR SCALE-UP AND INNOVATIONS AT IGV 24.3.1 Timeline and developmental steps 24.3.2 Industrial production PBRs in case descriptions 24.3.3 Main conclusions from experience to date 24.4 INNOVATIVE PROSPECTS 24.4.1 Light supply/layer thickness 24.4.2 O2/CO2 – concentration levels 24.4.3 Scale and economics ACKNOWLEDGMENT REFERENCES 25 Microalgae for Human and Animal Nutrition 25.1 INTRODUCTION 25.2 CHEMICAL COMPOSITION OF ALGAE 25.2.1 Protein 25.2.2 Amino acids 25.2.3 Carbohydrates 25.2.4 Lipids 25.2.5 Vitamins 25.2.6 Pigments 25.3 TOXICOLOGICAL ASPECTS 25.3.1 Biogenic toxins 25.3.2 Non-biogenic toxins 25.4 TOXICOLOGY 25.4.1 Toxicological studies with animals 25.4.2 Toxicological studies with humans 25.5 UTILIZATION OF ALGAL BIOMASS AND ALGAL CONSTITUENTS 25.5.1 Algae as animal feed 25.5.2 Therapeutic applications of algae 25.6 NUTRITIONAL QUALITY STANDARD AND PROCESS CONTROL REFERENCES 26 Bioactive and Novel Chemicals from Microalgae 26.1 INTRODUCTION 26.2 PRIMARY COPRODUCTS WITH POTENTIAL COMMERCIAL VALUE 26.2.1 β-carotene 26.2.2 Lycopene 26.2.3 Astaxanthin 26.2.4 Phycobiliproteins 26.3 SECONDARY METABOLITES WITH POTENTIAL COMMERCIAL VALUE 26.3.1 Antiviral activity 26.3.2 Ion channel modulatory activity 26.3.3 Immunomodulatory activity 26.3.4 Anticancer activity 26.4 NEW COMPOUND DISCOVERY 26.5 BIOSYNTHETIC INVESTIGATIONS 26.6 UNIQUELY MODIFIED LIPID PRODUCTS 26.6.1 Mono- and polyunsaturated fatty acids 26.6.2 Oxylipins 26.6.3 Hydrocarbons 26.6.4 Terpene-based products 26.7 BIOFUEL APPLICATIONS 26.7.1 Biosynthetic manipulations for biofuels 26.8 BIOPLASTICS 26.9 CONCLUSION ACKNOWLEDGMENT REFERENCES 27 High-value Recombinant Protein Production in Microalgae 27.1 INTRODUCTION 27.2 HIGH-VALUE RECOMBINANT PROTEINS PRODUCED IN MICROALGAE 27.3 GENETIC TRANSFORMATION OF MICROALGAE 27.4 STRATEGIES FOR MAINTAINING ACCUMULATION OF RECOMBINANT PROTEINS 27.5 CONCLUSIONS ACKNOWLEDGMENTS REFERENCES 28 Molecular and Cellular Mechanisms for Lipid Synthesis and Accumulation in Microalgae: Biotechnological Implications 28.1 INTRODUCTION TO LIPID BIOSYNTHESIS 28.1.1 Fatty acid biosynthesis 28.1.2 The Kennedy pathway 28.1.3 Galactolipid:galactolipid galactosyltransferase 28.1.4 Phospholipid:diacylglycerol acyltransferase 28.1.5 Conversion of starch to TAG 28.2 TAG SEQUESTRATION AND LB BIOGENESIS 28.2.1 Composition and structure of LBs 28.2.2 Biogenesis of LBs in eukaryotic microalgae 28.2.3 Functions of algal LBs 28.2.4 Interaction of LBs with other organelles 28.3 PHYSIOLOGICAL ROLE OF TAG SYNTHESIS AND LB BIOGENESIS 28.4 BIOTECHNOLOGICAL IMPLICATIONS 28.4.1 Enhanced TAG production through metabolic engineering 28.4.2 Enhanced biomass production 28.4.3 Lipase-based biochemical conversion 28.5 PERSPECTIVE REFERENCES 29 Biofuels from Microalgae 29.1 INTRODUCTION 29.2 EFFICIENT USE OF SUNLIGHT 29.3 REDUCTION OF ENERGY INPUT 29.4 SCARCITY OF RAW MATERIALS 29.5 LIPID ACCUMULATION 29.6 STRAIN IMPROVEMENT 29.7 SCALE-UP 29.8 BIOREFINERY 29.9 DESIGN SCENARIOS AND LIFE CYCLE ASSESSMENT 29.10 CONCLUSION ACKNOWLEDGMENTS REFERENCES Part 4 Water Pollution and Bioremediation by Microalgae 30 Eutrophication and Water Poisons 30.1 PARALYTIC SHELLFISH POISONING 30.2 OTHER NEUROTOXINS CAUSING SHELLFISH POISONING 30.3 DIARRHETIC SHELLFISH POISONING 30.4 AMNESIC SHELLFISH POISONING 30.5 CYANOTOXINS 30.6 REACTIVE OXYGEN SPECIES AND HAEMOLYTIC POISONINGS 30.7 OTHER POISONING 30.8 INTERACTIONS BETWEEN SYSTEM PHYSICS, BIOLOGY AND EUTROPHICATION 30.9 POISONINGS AND HEALTH 30.10 MANAGEMENT AND OPPORTUNITIES REFERENCES 31 Water Purification: Algae in Wastewater Oxidation Ponds 31.1 INTRODUCTION 31.2 PRINCIPLES OF OPERATING STABILIZATION PONDS 31.2.1 Biological equilibrium 31.2.2 Health considerations 31.3 INDUSTRIAL WASTEWATER 31.4 BIODIESEL 31.5 SUMMARY REFERENCES 32 Absorption and Adsorption of Heavy Metals by Microalgae 32.1 INTRODUCTION 32.2 HEAVY METALS IN THE AQUATIC SYSTEM 32.3 MICROALGAL RESPONSE TO HEAVY METALS STRESS 32.4 CURRENT TECHNIQUES FOR METALS DETOXIFICATION 32.4.1 Physicochemical approaches 32.4.2 Biological approaches 32.5 POTENTIAL APPLICATIONS OF MICROALGAE IN HEAVY METAL BIOREMEDIATION REFERENCES Part 5 Microalgae for Aquaculture 33 Microalgae for Aquaculture: The Current Global Situation and Future Trends 33.1 INTRODUCTION: AQUACULTURE – A RAPIDLY DEVELOPING ENTERPRISE 33.2 GREEN WATER AQUACULTURE 33.3 CLEAR WATER AQUACULTURE 33.3.1 Filtering mollusks 33.3.2 Shrimp 33.3.3 Fish 33.4 REFINING THE PRODUCTS OF AQUACULTURE 33.5 FORESEEABLE DEVELOPMENTS 33.6 FUTURE MICROALGAE PRODUCTIONS REFERENCES 34 Microalga for Aquaculture: Practical Implications 34.1 INTRODUCTION 34.2 CULTURING METHODS AT THE HATCHERY 34.2.1 Production cost in hatcheries 34.2.2 Outsourced algae 34.3 COMMERCIAL ALGAE 34.3.1 Commercial algae production methods 34.4 CONTAMINATION IN ALGAE CULTURES 34.4.1 Controlling contaminants 34.5 GREEN WATER TECHNIQUE 34.5.1 Non-algal substitutes for green water 34.6 FINFISH HATCHERIES 34.7 CRUSTACEAN HATCHERIES 34.7.1 Crabs 34.7.2 Shrimp 34.8 BIVALVE HATCHERIES 34.8.1 Oysters 34.8.2 Manila clams 34.8.3 Mercenaria clams 34.8.4 Algal production for bivalves 34.8.5 Commercial algal concentrates and bivalves 34.9 UNIVALVE HATCHERIES – ABALONE 34.10 SEA CUCUMBER HATCHERIES 34.10.1 Sea cucumber culture in China’s Shandong Province 34.11 ZOOPLANKTON CULTURE 34.11.1 Rotifers 34.11.2 Artemia 34.12 INTEGRATION OF AQUACULTURE WITH ALGACULTURE 34.13 MICROALGAE IN EXTENSIVE FRESHWATER PONDS 34.14 ALGAE CULTURE IN CHINA’S SHANDONG PROVINCE 34.14.1 Hatchery-produced algae 34.14.2 Commercially produced algae REFERENCES 35 Transgenic Marine Microalgae: A Value-Enhanced Fishmeal and Fish Oil Replacement 35.1 INTRODUCTION – THE PRESENT AND POTENTIAL NEEDS 35.1.1 Why transgenic domestication? 35.2 LIMITATIONS TO HAVING ALGAL MEAL AS A REPLACEMENT 35.2.1 Production engineering – culture and harvest 35.2.2 Problems in genetically engineering the domestication of marine algae 35.2.3 Need to integrate production systems engineering and genetic engineering 35.3 CHOICE OF ALGAE 35.4 GENETICALLY ENGINEERING ALGAL “PLATFORMS” 35.4.1 Resistance to contamination 35.4.2 Quorum sensing 35.4.3 Maximum growth rate – photosynthetic efficiency 35.4.4 Heat tolerance 35.4.5 An inability to grow in nature 35.5 ENHANCING THE VALUE/QUALITY OF ALGAE AS FISHMEAL/FISH OIL REPLACEMENTS 35.5.1 Enhancing digestibility of algae 35.5.2 Enhancing macronutrients 35.5.3 Replacing expensive or banned components in feed 35.5.4 Other therapeutics and growth regulators 35.6 CONCLUDING REMARKS ACKNOWLEDGMENTS REFERENCES 36 Microalgae for Aquaculture: Nutritional Aspects 36.1 INTRODUCTION 36.2 NUTRITIONAL FEATURES OF MICROALGAE FOR AQUACULTURE 36.2.1 Gross composition 36.2.2 Protein 36.2.3 Amino acid composition 36.2.4 Lipids 36.2.5 Vitamins 36.3 CAROTENOIDS IN AQUACULTURE 36.3.1 Astaxanthin 36.3.2 Utilization of astaxanthin in ornamental fish 36.3.3 Lutein 36.4 PRESERVED ALGAE 36.5 USAGE AND POTENTIAL OF SPIRULINA (ARTHROSPIRA) IN AQUACULTURE REFERENCES 37 The Enhancement of Marine Productivity for Climate Stabilization and Food Security 37.1 BACKGROUND 37.1.1 Introduction 37.1.2 Ocean carbon cycle 37.1.3 Food from the ocean 37.1.4 Climate stabilization 37.1.5 Acidification 37.1.6 The photic zone carbon stoichiometery 37.2 THE TECHNOLOGY 37.2.1 Ocean fertilization 37.2.2 Micro- and macronutrient fertilization 37.2.3 The manufacture of reactive nitrogen 37.2.4 Injection by fixed pipeline 37.2.5 Injection by ship 37.2.6 Carbon sequestration 37.2.7 Numerical models 37.2.8 Costs of fixing and delivering nitrogen 37.3 LEGAL AND SOCIETAL ISSUES 37.3.1 Legal constraints 37.3.2 Societal issues 37.3.3 Public outreach 37.4 ENVIRONMENTAL CONSIDERATIONS 37.4.1 Identified risk 37.4.2 Unidentified risk 37.5 CONCLUSION REFERENCES Index