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Green Energy and Technology Jianlong Wang Yanan Yin Biohydrogen Production from Organic Wastes Green Energy and Technology More information about this series at http://www.springer.com/series/8059 Jianlong Wang Yanan Yin (cid:129) Biohydrogen Production from Organic Wastes 123 Jianlong Wang Yanan Yin Institute of Nuclear andNewEnergy Institute of Nuclear andNewEnergy Technology Technology TsinghuaUniversity TsinghuaUniversity Beijing Beijing China China ISSN 1865-3529 ISSN 1865-3537 (electronic) Green Energy andTechnology ISBN978-981-10-4674-2 ISBN978-981-10-4675-9 (eBook) DOI 10.1007/978-981-10-4675-9 LibraryofCongressControlNumber:2017939533 ©SpringerNatureSingaporePteLtd.2017 Thisworkissubjecttocopyright.AllrightsarereservedbythePublisher,whetherthewholeorpart of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission orinformationstorageandretrieval,electronicadaptation,computersoftware,orbysimilarordissimilar methodologynowknownorhereafterdeveloped. The use of general descriptive names, registered names, trademarks, service marks, etc. in this publicationdoesnotimply,evenintheabsenceofaspecificstatement,thatsuchnamesareexemptfrom therelevantprotectivelawsandregulationsandthereforefreeforgeneraluse. The publisher, the authors and the editors are safe to assume that the advice and information in this book are believed to be true and accurate at the date of publication. Neither the publisher nor the authorsortheeditorsgiveawarranty,expressorimplied,withrespecttothematerialcontainedhereinor for any errors or omissions that may have been made. The publisher remains neutral with regard to jurisdictionalclaimsinpublishedmapsandinstitutionalaffiliations. Printedonacid-freepaper ThisSpringerimprintispublishedbySpringerNature TheregisteredcompanyisSpringerNatureSingaporePteLtd. Theregisteredcompanyaddressis:152BeachRoad,#21-01/04GatewayEast,Singapore189721,Singapore Preface The environment pollution and energy crisis are two vital issues for sustainable developmentworldwide.Theuseoffossilfuels(petroleum,naturalgas,andcoal)is considered as the cause of serious environmental problems. With the increasing demand of energy and depleting reserves of conventional fossilfuels,therehasbeengrowingglobalinterestindevelopingalternativesources of energy. Although hydrogen (H ) is not a primary energy source, it has been 2 considered a promising candidate as a substitute for fossil fuels because it has the potential to eliminate most of the problems caused by the fossil fuels. Fromtheperspectiveofenergysource,hydrogencanbeproducedfromorganic wastes (agricultural wastes, municipal wastes, algal biomass, etc.), renewable energy(solar,wind,etc.),fossilfuel(coal,oil,naturalgas,etc.),andnuclearenergy. Different technologies have been used for hydrogen production, such as direct thermal, thermochemical (such as hydrocarbon reforming and coal gasification), electrochemical, and biological. Among the hydrogen production methods, hydrogenproductionusingbiologicalprocessesisnew,innovative,andpotentially more efficient, which has been broadly studied for its mild reaction condition and high potential environmental benefits. Biohydrogen production is performed by hydrogen-producing microorganisms at ambient temperature and pressure. Microorganisms can recover and concentrate theenergyfromaqueousorganicwastes,suchasindustrialwastewater andsludge. Biohydrogen can be produced from the biophotolysis of water using algae and cyanobacteria, the photo-decomposition of organic compounds by photosynthetic bacteria, and the dark fermentation from organic compounds with anaerobic bac- teria. Among these biological processes, dark fermentation is more favorable than photo-dependenthydrogenproductionforitsindependencyoflight,generallyhigh rate of hydrogen generation, simple reactor as well as easy control. In particular considering the wide range of substrates, dual benefits of clean energy generation andorganicwastesmanagementcanbeachieved,sincehydrogenisproducedfrom various organic wastes and wastewater enriched with carbohydrates as the sub- strate, decreasing the cost for hydrogen production. Thus, fermentative hydrogen v vi Preface productioniswidelyacceptedasamorefeasiblebiohydrogenproductionway,and gained widespread interest and attention. Biological hydrogen production processes offer a technique through which renewable energy sources like biomass can be utilized for the generation of the cleanestenergycarrier.Hydrogen-intensiveresearchworkhasalreadybeencarried outontheadvancementoftheseprocesses,suchasthedevelopment ofgenetically modified microorganism, metabolic engineering, improvement of the reactor designs, use of different solid matrices for the immobilization of whole cells, biochemical-assistedbioreactor,anddevelopmentoftwo-stageprocessesforhigher hydrogen production rate. The present book provides the state-of-the-art information on the status of the biohydrogenproductionfromvariousorganicwastes.Thisbookhaseightchapters, including the microbiology, biochemistry and enzymology of biohydrogen pro- duction, the enrichment of hydrogen-producing microorganisms, the pretreatment of various organic wastes for hydrogen production, the influence of different physicochemicalfactorsonhydrogenproduction,thekineticmodelsandsimulation of biological process of fermentative hydrogen production, the optimization of biologicalhydrogenproductionprocess,andthefermentativehydrogenproduction from sewage sludge. The text in all the chapters is supported by numerous clear, informative figures andtables.Toourknowledge,thisbookisafirstattempttodescribethebiological hydrogen production from various organic wastes, which is aimed at a wide range of readers, mainly including undergraduates, postgraduates, energy researchers engineers, and others who are interested in hydrogen production in general and biological hydrogen production in particular, as well as to industrial concerns that are looking for inexpensive hydrogen production technologies. We warmly thank Dr. Mengchu Huang and Dr. Sivajothi Ganesarathinam, and theteamofSpringerNaturefortheircooperationandeffortsinproducingthisbook. Wehopereaderswillfindthisbookinterestingandinformativefortheirresearch pursuits. Beijing, China Jianlong Wang Yanan Yin Contents 1 Introduction . .... .... .... ..... .... .... .... .... .... ..... .. 1 1.1 Global Energy Development . .... .... .... .... .... ..... .. 1 1.2 Hydrogen Energy. .... ..... .... .... .... .... .... ..... .. 3 1.3 Hydrogen Production.. ..... .... .... .... .... .... ..... .. 4 1.3.1 Steam Reforming ... .... .... .... .... .... ..... .. 5 1.3.2 Electrolysis... ..... .... .... .... .... .... ..... .. 5 1.3.3 Thermal Chemical... .... .... .... .... .... ..... .. 6 1.3.4 Biological.... ..... .... .... .... .... .... ..... .. 6 1.4 Overview of This Book..... .... .... .... .... .... ..... .. 15 References ... .... .... .... ..... .... .... .... .... .... ..... .. 16 2 Microbiology and Enzymology ... .... .... .... .... .... ..... .. 19 2.1 Microorganisms in Hydrogen-Producing System.. .... ..... .. 19 2.1.1 Overview .... ..... .... .... .... .... .... ..... .. 19 2.1.2 Microbial Diversity in Hydrogen-Producing System.. .. 20 2.2 Inocula for Dark Fermentation.... .... .... .... .... ..... .. 20 2.2.1 Mixed Culture. ..... .... .... .... .... .... ..... .. 21 2.2.2 Pure Culture.. ..... .... .... .... .... .... ..... .. 22 2.3 Pure Culture for Hydrogen Production.. .... .... .... ..... .. 25 2.3.1 Clostridium butyricum INET1 . .... .... .... ..... .. 25 2.3.2 Enterococcus faecium INET2.. .... .... .... ..... .. 34 2.4 Biochemistry of Hydrogen Production.. .... .... .... ..... .. 46 2.4.1 Metabolic Pathways . .... .... .... .... .... ..... .. 46 2.4.2 Fermentation Types . .... .... .... .... .... ..... .. 48 2.5 Enzymology of Hydrogen Production .. .... .... .... ..... .. 49 2.5.1 Classification of Hydrogenase.. .... .... .... ..... .. 50 2.5.2 Genetic Modification of Hydrogenase.... .... ..... .. 54 2.5.3 Environmental Applications of Hydrogenase .. ..... .. 55 2.6 Microbial Modification. ..... .... .... .... .... .... ..... .. 56 2.6.1 Co-cultivation. ..... .... .... .... .... .... ..... .. 56 vii viii Contents 2.6.2 Microbial Immobilization . .... .... .... .... ..... .. 58 2.6.3 Metabolic Engineering ... .... .... .... .... ..... .. 59 References ... .... .... .... ..... .... .... .... .... .... ..... .. 61 3 Enrichment of Hydrogen-Producing Microorganisms . .... ..... .. 69 3.1 Overview... .... .... ..... .... .... .... .... .... ..... .. 69 3.2 Heat Treatment .. .... ..... .... .... .... .... .... ..... .. 70 3.3 Acid/Alkaline Treatment .... .... .... .... .... .... ..... .. 74 3.4 Chemical Inhibitors ... ..... .... .... .... .... .... ..... .. 76 3.5 Aeration.... .... .... ..... .... .... .... .... .... ..... .. 76 3.6 Other Treatments. .... ..... .... .... .... .... .... ..... .. 78 3.6.1 Ultrasonication ..... .... .... .... .... .... ..... .. 78 3.6.2 Freezing and Thawing ... .... .... .... .... ..... .. 80 3.6.3 Electric Treatment... .... .... .... .... .... ..... .. 81 3.6.4 Microwave Treatment.... .... .... .... .... ..... .. 83 3.6.5 Ionizing Radiation Treatment .. .... .... .... ..... .. 85 3.6.6 Ultraviolet (UV) Radiation.... .... .... .... ..... .. 86 3.6.7 Load-Shock Treatment ... .... .... .... .... ..... .. 86 3.6.8 Operational Condition Control . .... .... .... ..... .. 86 3.7 Combined Treatments . ..... .... .... .... .... .... ..... .. 87 3.8 Effect of Pretreatment Methods on Microbial Community.... .. 89 3.9 Comparison of Different Pretreatment Methods... .... ..... .. 91 3.10 Gamma Irradiation for Enriching Hydrogen-Producer .. ..... .. 93 3.10.1 Overview .... ..... .... .... .... .... .... ..... .. 93 3.10.2 Effect of Dose on Hydrogen Production.. .... ..... .. 94 3.10.3 EffectofDoseonSubstrateDegradationandHydrogen Yield.... .... ..... .... .... .... .... .... ..... .. 96 3.10.4 Effect of Dose OnVolatile Fatty Acids... .... ..... .. 97 3.10.5 Conclusions .. ..... .... .... .... .... .... ..... .. 99 3.11 Hydrogen Production Performance by Different Pretreated Sludge . .... .... .... ..... .... .... .... .... .... ..... .. 100 3.11.1 Effect on Hydrogen Production. .... .... .... ..... .. 100 3.11.2 Effect on Substrate Degradation and Hydrogen Yield. .. 102 3.11.3 Effect on Volatile Fatty Acids and Final pH... ..... .. 104 3.12 Changes in Microbial Community During Biohydrogen Production .. .... .... ..... .... .... .... .... .... ..... .. 107 3.12.1 Seed Sludge and Fermentation Conditions .... ..... .. 107 3.12.2 DNA Extraction and PCR Amplification . .... ..... .. 107 3.12.3 MiSeq Sequencing and Data Analysis ... .... ..... .. 107 3.12.4 Hydrogen Production Progress . .... .... .... ..... .. 108 3.12.5 Microbial Diversity Characteristics.. .... .... ..... .. 109 3.12.6 Microbial Diversity at Different Stages... .... ..... .. 111 References ... .... .... .... ..... .... .... .... .... .... ..... .. 113 Contents ix 4 Pretreatment of Organic Wastes for Hydrogen Production. ..... .. 123 4.1 Overview... .... .... ..... .... .... .... .... .... ..... .. 123 4.2 Main Structural Components of Organic Wastes .. .... ..... .. 124 4.3 Types and Compositions of Organic Wastes . .... .... ..... .. 124 4.3.1 Waste Activated Sludge .. .... .... .... .... ..... .. 127 4.3.2 Algal Biomass ..... .... .... .... .... .... ..... .. 128 4.3.3 Cellulose-Based Biomass . .... .... .... .... ..... .. 131 4.3.4 Starch-Based Biomass.... .... .... .... .... ..... .. 134 4.3.5 Food Waste... ..... .... .... .... .... .... ..... .. 134 4.3.6 Wastewater... ..... .... .... .... .... .... ..... .. 136 4.4 Pretreatment of Organic Wastes... .... .... .... .... ..... .. 138 4.4.1 Physical Treatment.. .... .... .... .... .... ..... .. 138 4.4.2 Chemical Treatment . .... .... .... .... .... ..... .. 145 4.4.3 Biological Treatment. .... .... .... .... .... ..... .. 147 4.4.4 Combined Treatment. .... .... .... .... .... ..... .. 148 4.4.5 Comparison of Different Treatment Methods .. ..... .. 149 4.5 Hydrogen Production from Organic Wastes.. .... .... ..... .. 152 4.5.1 Hydrogen Production from Waste Activated Sludge.. .. 152 4.5.2 Hydrogen Production from Algal Biomass.... ..... .. 152 4.5.3 Hydrogen Production from Cellulose-Based Biomass. .. 165 4.5.4 Hydrogen Production from Starch-Based Biomass ... .. 175 4.5.5 Hydrogen Production from Food Wastes . .... ..... .. 175 4.5.6 Hydrogen Production from Wastewater .. .... ..... .. 177 4.6 Concluding Remarks and Perspectives.. .... .... .... ..... .. 181 References ... .... .... .... ..... .... .... .... .... .... ..... .. 183 5 Influencing Factors for Biohydrogen Production. .... .... ..... .. 197 5.1 Introduction. .... .... ..... .... .... .... .... .... ..... .. 197 5.2 Effect of Inoculum.... ..... .... .... .... .... .... ..... .. 198 5.2.1 Pure Cultures . ..... .... .... .... .... .... ..... .. 198 5.2.2 Mixed Cultures..... .... .... .... .... .... ..... .. 198 5.3 Effect of Substrate .... ..... .... .... .... .... .... ..... .. 203 5.3.1 Overview .... ..... .... .... .... .... .... ..... .. 203 5.3.2 Effect on Substrate Degradation Efficiency.... ..... .. 208 5.3.3 Effect on Hydrogen Production. .... .... .... ..... .. 209 5.3.4 Effect on Hydrogen Production Rate. .... .... ..... .. 210 5.3.5 Effect on Soluble Metabolites Distribution.... ..... .. 211 5.3.6 Effect on Final pH .. .... .... .... .... .... ..... .. 212 5.4 Effect of Reactor Type. ..... .... .... .... .... .... ..... .. 213 5.5 Effect of Nitrogen and Phosphate.. .... .... .... .... ..... .. 216 5.5.1 Overview .... ..... .... .... .... .... .... ..... .. 216 5.5.2 Effect of Ammonia Concentration... .... .... ..... .. 219 5.5.3 Effect of Nitrate Concentration. .... .... .... ..... .. 224

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