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189 Pages·2011·6.39 MB·English

ANAEROBIC DIGESTION OF BIOFUEL PRODUCTION RESIDUES

By

ZHUOLI TIAN

A DISSERTATION PRESENTED TO THE GRADUATE SCHOOL
OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT
OF THE REQUIREMENTS FOR THE DEGREE OF
DOCTOR OF PHILOSOPHY

UNIVERSITY OF FLORIDA

2011

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© 2011 Zhuoli Tian

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To my parents who have always been supportive of my education

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ACKNOWLEDGMENTS
I would like to extend my deepest appreciation to my parents, Mr.Tian and Mrs.
Zheng for their continuous support and encouragement towards my academic
endeavors.
I express my sincere gratitude to my advisor and committee chairman, Dr. Pratap
Pullammanappallil, for his continual support and guidance during my time at the
University of Florida. I am grateful to Dr. Lonnie O. Ingram for his support throughout
this research study. I would like to thank Dr. Ben Koopman and Dr. Paul A. Chadik for
serving on my committee and their inspirational lectures. I also own gratitude to Dr.
Melanie Correll for her devotion and patience in helping improve my writing skills. I give
thanks to Dr. Adriana Giongo and Ms. Diane Chauliac for their guidance and assistance
in carrying out microbial anaysis for this research. I also would like to thank Dr. Claudia
C. Geddes and Dr. Mike Mullinnix for providing necessary materials and data for this
research, as well as their insightful ideas and assistances in solving analytical issues.
I would like to thank the following individuals for their supports and assistances
during my studies, Abhay Koppar, Cesar Moreira , Congrong Yu, Gayathri Ram Mohan,
Ioannis M. Polematidis, Jaime Chavez Leon, Mandu Inyang, Patrick Dube, Samriddhi
Buxy, Yuan Tian and all the friends in the Department of Agricultural and Biological
Engineering.
I gave my special thanks to Lushun Xiang and Ying Yao for their wholehearted
supports and companies to relieve my frustration and stress. This would never have
been possible without them.

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TABLE OF CONTENTS

page
ACKNOWLEDGMENTS..................................................................................................4
LIST OF TABLES............................................................................................................8
LIST OF FIGURES..........................................................................................................9
ABSTRACT...................................................................................................................11
CHAPTER
1 INTRODUCTION....................................................................................................14
Project Focus..........................................................................................................14
Glycerol Byproduct from Biodiesel Production Process..........................................15
Residues from Bioethanol Production.....................................................................16
Research Objectives...............................................................................................18
Anaerobic Digestion Technique..............................................................................19
The Microbiology of Anaerobic Digestion.........................................................19
Methane Production and COD Degradation.....................................................21
Configurations of Anaerobic Digesters.............................................................22
Factors Affecting Anaerobic Digestion .............................................................23
Methods for Investigation of Microbial Communities........................................25
Current Technologies for 16S rDNA Sequencing.............................................25
16S rDNA Sequencing Data Analysis ..............................................................26
2 BIOCHEMICAL METHANE POTENTIAL OF GLYCEROL BYPRODUCT FROM
BIODIESEL PRODUCTION USING WASTE VEGETABLE OIL.............................31
Summary................................................................................................................31
Background.............................................................................................................32
Material and Method...............................................................................................33
Feedstock.........................................................................................................33
BMP Assay.......................................................................................................34
Semi-continouous Digestion of GBP................................................................35
Analysis............................................................................................................36
Results....................................................................................................................38
Characterization of the GBP.............................................................................38
Methane Potential ............................................................................................38
Soluble Chemical Oxygen Demand Profiles.....................................................42
Volatile Organic Acid Profiles...........................................................................43
Methane Production from Semi-continuous Digestion of GBP.........................43
Discussion........................................................................................................45
Closing Remarks ....................................................................................................48
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3 ANAEROBIC DIGESTION FOR TREATMENT OF STILLAGE FROM
CELLULOSIC BIOETHANOL PRODUCTION ........................................................55
Summary................................................................................................................55
Background.............................................................................................................56
Materials and Methods............................................................................................58
Feedstock.........................................................................................................58
Biochemcial Methane Potential Assay (BMP)..................................................59
Continuous Digestion of Bagasse Stillage Filtrate............................................60
Monitoring and Analysis...................................................................................62
Results....................................................................................................................64
Characterization of the Stillage.........................................................................64
Methane Potential ............................................................................................65
Soluble COD and Volatile Organic Acids in BMP Assays ................................66
Continuous Digestion of Bagasse Stillage Filtrate (BF)....................................67
Discussion........................................................................................................68
Closing Remarks ....................................................................................................75
4 COMPARISON OF BATCH ANAEROBIC DIGESTION OF SUGARBEET
TAILINGS AT MIXED AND NON-MIXED CONDITIONS........................................88
Summary................................................................................................................88
Background.............................................................................................................89
Material and Method...............................................................................................91
Feedstock.........................................................................................................91
Anaerobic Digesters.........................................................................................91
Experiment Description....................................................................................92
Temperature Monitor........................................................................................93
Chemical Analysis............................................................................................94
Microbial Community Analysis..........................................................................94
Results....................................................................................................................96
Characterization of Substrates.........................................................................96
Temperature Profiles........................................................................................96
CH Production.................................................................................................97
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Profiles of sCOD Degradation Comparison......................................................98
Profiles of Volatile Organic Acid.......................................................................98
Microbial Communities Structure......................................................................99
Discussion............................................................................................................100
Determination of Mixing Intensity...................................................................100
Digestion Performance Comparison...............................................................107
Microbial Community......................................................................................108
COD Balance.................................................................................................111
Closing Remarks ..................................................................................................113
5 MICROBIAL POPULATION IDENTIFICATION AND DYNAMICS DURING
BATCH THERMOPHILIC ANAEROBIC DIGESTION AT MIXED AND NON-
MIXED CONDITIONS...........................................................................................126
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Summary..............................................................................................................126
Background...........................................................................................................127
Method and Materials...........................................................................................128
Feedstock.......................................................................................................128
Anaerobic Reactors and Operation................................................................128
Chemical Analysis..........................................................................................129
Molecular Biological Analysis.........................................................................129
DNA Extraction and Purification.....................................................................130
Polymerase Chain Reaction Amplification......................................................130
Results..................................................................................................................132
Characteristics of Feed Substrates ................................................................132
Methane Production.......................................................................................132
Degradation of Organic Matters .....................................................................133
Microbial Community Analysis........................................................................134
Discussion............................................................................................................136
Digestion Performance Comparison...............................................................136
Microbial Community......................................................................................137
Closing Remarks ..................................................................................................147
6 CONCLUSIONS AND FUTURE WORK...............................................................158
Conclusions..........................................................................................................158
Future Work..........................................................................................................162

APPENDIX
A EMERGY ANALYSIS OF BIODIESEL PRODCTION FROM WASTE
VEGETABLE OIL..................................................................................................163
B DEGRADATION OF PROPIONIC ACID IN MIXED ANAEROBIC DIGESTION ...164
LIST OF REFERENCES.............................................................................................170
BIOGRAPHICAL SKETCH..........................................................................................189

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LIST OF TABLES
Table page

1-1 Comparison of Anaerobic Digesters...................................................................27
2-1 Contents and constituents of the assays............................................................50
2-2 Paramerers of Gompertz Equation.....................................................................50
3-1 Contents and constituents of the BMP assays...................................................77
3-2 Characteristics of bagasse whole stillage...........................................................78
3-3 Monitored parameters of anaerobic digestion ....................................................79
4-1 Substrate characteristics and loading amounts for digester 1 and 2................114
4-2 Power input per unit volume.............................................................................115
4-3 Bacterial 16S rRNA gene library.......................................................................116
4-4 Archaeal 16S rRNA gene library.......................................................................117
5-1 PCR amplifications of 16s rRNA genes using 12 barcoded primers.................149
5-2 Substrate characteristics and loaded quantities for digesters 1 and 2..............150
5-3 Analysis of 16S-rDNA sequence at taxon rank of order...................................151
A-1 Emergy evaluation of biodiesel production.......................................................163

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LIST OF FIGURES
Figure page

1-1 Transesterification of triglyceride........................................................................29
1-2 Ethanol production process from sugarcane bagasse........................................30
2-1 Cumulative methane yield from assay A-1, C-II, 1 and A-3................................51
2-2 sCOD profile in assays A-1, A-2, C-II, 1 and C-II, 2............................................52
2-3 Propionic acid profiles from assay A-1 and A-2..................................................53
2-4 Daily methane yields from semi-continuous digestion of GBP ...........................54
3-1 Custom-built tripod stand for anaerobic digesters..............................................80
3-2 Continuous anaerobic reactor schematic ...........................................................81
3-3 Methane yields of Assay BW-1, BW-2, BF, BR, HW-1 and HW-2......................82
3-4 COD and VOA profiles of assay BW-1 and BW-2..............................................83
3-5 COD and VOA profiles of assay HW-1 and HW-2..............................................84
3-6 Daily methane yield from continuous digestion of stillage ..................................85
3-7 Profiles of SCOD, VOA, phosphorus and ammonia as N...................................86
3-8 Mass balance of the integrated process.............................................................87
4-1 Digester and gas meter configuration...............................................................118
4-2 Inoculum used for experimental set 1 and 2.....................................................119
4-3 Temperature profiles of incubator, digester 1 and digester 2 ...........................120
4-4 Methane yield in digester 1 and 2 from all 6 runs.............................................121
4-5 SCOD concentration in digester 1 and 2 from 6 runs.......................................122
4-6 VOA concentration in digester 1 and 2 from three runs....................................123
4-7 The rank abundances of bacterial and archaeal phylotypes.............................124
4-8 Average COD balance of digesters 1 and 2.....................................................125
5-1 Methane yield from digesters 1 and 2...............................................................153
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5-2 SOCD and VOA profiles of digester 1 and digester 2.......................................154
5-3 Estimate of Chao1 coverage............................................................................155
5-4 Microbial community dynamics of digester 1 and 2..........................................157
B-1 pH profile..........................................................................................................168
B-2 VOA profile.......................................................................................................169

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