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研究生：	梁德明 Liang, Teh-Ming
論文名稱：	薄膜分離應用於厭氧生物產氫程序 Application of membrane separation on anaerobic hydrogen-producing processes
指導教授：	鄭幸雄 Cheng, Sheng-Shung
學位類別：	博士 Doctor
系所名稱：	工學院 - 環境工程學系 Department of Environmental Engineering
論文出版年：	2004
畢業學年度：	92
語文別：	英文
論文頁數：	193
中文關鍵詞：	薄膜分離、產氫生成率、生物產氫速率、厭氧生物產氫、全蒸發薄膜、微過濾膜、電透析膜
外文關鍵詞：	Anaerobic hydrogen-producing processes, hydrogen yield, hydrogen-producing rate, pervaporation membrane, membrane separation, microfiltration membrane, electrodialysis membrane
相關次數：	點閱：102 下載：1
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本研究涵蓋厭氧生物對葡萄糖(glucose)與蛋白胨(peptone)產氫動力探討，醋酸與溫度對厭氧生物產氫的影響，與啟動厭氧生物產氫發酵槽，並探討三種不同薄膜對厭氧產氫發酵槽操作的影響。三種薄膜包括:以全蒸發膜分離氫氣，降低醱酵槽氫分壓以促進產氫，以微過濾膜回收產氫菌增加產氫發酵槽的功能；以電透析膜分離醋酸與氨氮降低抑制性，利於厭氧生物產氫，同時減少醱酵液中氨氮對後續厭氧光合產氫菌的抑制，進一步被厭氧光合產氫菌利用產生氫氣。
氫氣、二氧化碳、乙酸、丁酸與乙醇是厭氧氫醱酵的主要產物，過度累積將減緩產氫效率與產氫菌生長，如能快速將生成物分離將有助於生物產氫。在批次厭氧產氫試驗，基質產氫率與產物生成的比例受基質與微生物濃度 (So/Xo) 比值的影響。當 So/Xo 比值從 5 增加到333 g COD/g VSS，醱酵產物中的丙酸與甲酸減少, 但是乙酸、丁酸與乙醇增加。使用強化產氫培養的污泥對葡萄糖的產氫率比使用熱處理的後的產氫污泥為高，產氫速率與基質濃度有關，呈現Monod 經驗式的關係。同樣對peptone 進行批次厭氧產氫試驗，單位peptone的產氫率較葡萄糖為低，但是微生物增殖率較葡萄糖為高。
研究乙酸對厭氧生物產氫的影響發現，在中性pH下乙酸高達2,500 mg/L，只有輕微的影響產氫量與產氫速率。當pH在5時，乙酸非常明顯產生抑制作用降低基質產氫率與減少產氫速率。使用吸附劑(γ-alumina) 可以減少溶液中分子態乙酸增加產氫速率。在溫度影響方面，高溫厭氧產槽(thermophilic anaerobic hydrogen fermentor , TAHF, 55±2 oC)的基質產氫率為 8.4 mmol H2/g COD ，高於中溫厭氧氫醱酵槽(mesophilic anaerobic hydrogen fermentation, MAHF, 35±2 oC)的基質產氫率 5.6 mmol H2/g COD。高溫厭氧產槽的比產氫速率(specific H2-producing rate) 為7.1 mmol H2/g VSS-hr 也高於中溫厭氧氫醱酵槽的比產氫速率(5.8 mmol H2/g VSS-hr)。中溫厭氧醱酵液相產物以乙酸與丁酸居多，而高溫厭氧醱酵以乙醇與乙酸居多。
研究啟動厭氧產氫槽，以厭氧產甲烷污泥床的顆粒污泥為植種源，在35oC批次培養中添加 28 mg/L 的氯仿(chloroform)於植種污泥中，成功抑制甲烷菌活性，基質產氫率為1.48 mmol H2 /g peptone。因此，可使用氯仿經由批次或半批次培養啟動產氫槽。厭氧產甲烷污泥床的顆粒污泥也是具有很好產氫活性的植種菌源。
將產氫反應槽裝設全蒸發薄膜(pervaporation membrane)，可迅速將氫氣分離。氣體的質傳係數與薄膜之cross-flow velocity成正比，亦即利用中空纖維膜的表面切線速度可以增加質傳效率。當斷面流速增加, CO2質傳係數由0.81×10-7 增加至 3.19×10-7 cm/sec-KPa， H2 的質傳係數由0.55×10-8 升高至1.96×10-8 cm/sec-KPa。就矽膠膜的質傳特性而言，二氧化碳質傳速率大於氫氣的質傳速率。實驗結果顯示，降低H2與CO2的分壓增加10% 的產氫速率與15%.的基質產氫率。
研究以微過濾薄膜(microfiltration membrane)進行高溫厭氧產氫槽回收產氫菌體，薄膜反應槽確實能達到污泥截留之功能，有效提昇反應槽中污泥濃度由 500 mg/L 增加至 5,200 mg VSS/L。厭氧產氫槽採用混合菌種發酵蔗糖與蛋白胨的複合基質，在55oC與體積負荷80 g COD/L-day條件下，反應槽最大的產氫為28 mmol H2/L-hr，比產氫速率為4.6 mmol H2/g VSS-hr，基質產氫率為8.41 mmol H2/g COD。當食微比在4 到 16 g COD/g VSS-day之間，產氫速率與食微比成正比。厭氧產氫槽體積負荷由20 升高至 80 g COD/L-day時，丁酸濃度也從50 升高至 2,800 mg/L。雖然高污泥量會降低產氫速率，但對於反應槽的穩定卻是有幫助的，這也是薄膜反應槽優於CSTR 反應槽的優點之一。
利用電透析的原理將厭氧產生成的產物(包含揮發酸與氨)，藉帶電荷不同經由陰陽離子交換膜，予以分離，可以降低發酵液體中產物抑制的情形。研究顯示在電透析反應槽中不同的停留時間與端電壓，影響液相中有機酸與氨氮的分離效果。當HRT愈長分離效果愈好，在兩小時的HRT已經有70%的分離效果。同樣的電壓愈高分離效果也愈好。因此以電透析膜分離醱酵產物中的乙酸與氨氮是具有可行性。

This research includes to investigate biokinetics of hydrogen fermentation in glucose and peptone and effects of acetate and temperature in hydrogen fermentation, and to start up a fermentative hydrogen reactor using a granular sludge as seeding sludge, as well as to investigate effects of three types of membranes installed with a hydrogen fermentor. The membranes include a pervaporation membrane to separate H2 to reduce hydrogen partial pressure, and a microfiltration membrane to recover biomass, and an electrodialysis membrane to separate fermented products.
Hydrogen, volatile acids, ammonium and alcohol are products of hydrogen fermentation. The products will retard the growth and metabolism of cells to decrease hydrogen production, once they accumulate in the reactor. If they can be separated immediately, hydrogen production can increase significantly. Hydrogen yield is also affected by the ratio of substrate and biomass concentrations in batch fermentation of glucose. The portions of fermentation products are also varied with the ratios. When the So/Xo escalates from 5 to 333, the portions of propionate and formate decrease, but the acetate, butyrate and ethanol increase. Hydrogen yields of glucose fermentation using enhanced sludge exceed that of using preheated sludge. Hydrogen-producing rates are correlated with the concentrations of glucose, and hence they can be depictured with Monod equation. Hydrogen fermentation of peptone has a less hydrogen yield but a higher growth yield than glucose based on one gram of substrate.
Investigation of factors affecting anaerobic hydrogen production shows that acetate affects biohydrogen production slightly at neutral pH, but it significantly depresses the hydrogen-producing rate and hydrogen yield at pH 5. To useγ-alumina as an absorbent in serum bottles to decrease volatile acids in bulk solution can enhance hydrogen-producing rate and yield for peptone and. However, to remove the acids of bulk solution immediately could promote the hydrogen production rates. The thermophilic anaerobic hydrogen fermentor (TAHF, 55±2 oC ) obtained a hydrogen yield of 8.4 mmol H2/g COD larger than 5.6 mmol H2/g COD of the mesophilic anaerobic hydrogen fermentation (MAHF, 35±2 oC). The specific H2-producing rate of 7.1 mmol H2/g VSS-hr of the TAHF bioreactor exceeds 5.8 mmol H2/g VSS-hr of MAHF. The recovery of electron flow is 88.7% in converting one gram of substrate in the TAHF bioreactor, and 73.2 % in the MAHF bioreactor. Acetic acid and butyric acid predominated in the MAHF bioreactor, but ethanol and acetic acid predominated in the TAHF. The thermophilic bioreactor achieves higher specific H2-producting rates and hydrogen yields.
Investigating hydrogen production of granular sludge via inhibition of methanogensis shows that 28 mg/L of chloroform in the mixed liquor of 10 g VSS/L was effective in inhibiting methanogenic activity in batch incubation at 35oC. No methane was produced from the bottle with adding of chloroform and the hydrogen yield was 1.48 mmol H2 /g peptone. Batch incubation with chloroform and semi-batch running for start-up might demolish the methanogens. Consequently, the vital anaerobic granular sludge from a UASB reactor can be used as a good source of hydrogen-fermentation consortia.
This investigation examines the effectiveness of a silicone rubber membrane to separate biogas from the liquid medium in the hydrogen fermentation reactor. When the culture liquid circulates through the shell side of the hollow fibers, the biogas diffuses into the lumen of the hollow fibers and is removed by a vacuum pump. When the cross flow velocity along the module increases, the CO2 mass transfer coefficient increases from 0.81×10-7 to 3.19×10-7 cm/sec-KPa, and the H2 mass transfer coefficient increases from 0.55×10-8 to 1.96×10-8 cm/sec-KPa. The mass transfer coefficients of CO2 and H2 are directly proportional to the cross flow velocity. However, silicone rubber effectively reduces the biogas partial pressure in the hydrogen fermentor, and improves the hydrogen- producing rate by 10% and the hydrogen yield by 15%.
Using a microfiltration membrane to recover cells of hydrogen-producing bacteria succeeds to accumulate biomass from 500 mg/L to 5,200 mg VSS/L. The fermentor employed mix-cultured consortia to ferment multiple substrate including peptone and glucose at 55oC. The maximum H2-producing rate is 28 mmol H2/L-hr and the maximum specific H2-producing rate is 4.6 mmol H2/g VSS-hr when the fermentor was operated at more than 80 g COD/L-day. The hydrogen yield is 8.41 mmol H2/g COD. Excepting H2, the other fermented products included acetic acid, butyric acid, and ethanol. The concentration of butyric acid increased from 50 to 2,800 mg/L when the loading increased from 20 to 80 g COD/L-day. The results show that specific H2-producing rates directly proportion to F/M ratios from 4 to 8 g COD/g VSS-day.
Using an electrodialysis membrane succeeds to separate acetate and ammonium. The removal of acetate reaches 70% at a HRT of 2 hr. When the HRT is longer than 2 hr, it will attain higher efficiency in separating acetate as well as ammonium. The voltage of the electrodialysis cell significantly determines the efficiency of separation, too. High voltage achieves a high efficiency of separation. The parameters of HRT and voltage govern the separation, and the results shows the possibilities of separation acetate and ammonium with an electrodialysis membrane.

Contents
Abstract (Chinese)	I
Abstract	IV
Contents	VII
List of Figures	XII
List of Tables	XVI
Chapter 1 Introduction	1
1-1 Motivation	1
1-2 Objectives	3
1-3 Scope	         3
Chapter 2 Literature review	                  6
2-1 Biological hydrogen production	         6
2-1-1 Photosynthetic hydrogen production	10
2-1-2 Fermentative hydrogen production	12
2-2 Mechanism of fermentative hydrogen production	         15
2-2-1 Thermodynamic in fermentative hydrogen production	15
2-2-2 Fermentative hydrogen production from carbohydrate	19
2-2-3 Fermentative hydrogen production from protein	         23
2-2-4 Anaerobic hydrogen production from multiple substrate	26
2-2-5 Anaerobic hydrogen production from wastes	         26
2-3 Microbiology of fermentative hydrogen production            28
2-3-1 Clostridium	                                              29
2-3-2 Enteroaerogens	                                     31
2-3-3 Isolation and identification of hydrogen producing bacteria 33
2-4 Kinetics of fermentative hydrogen production                 37
2-5 Effects of environmental factors on fermentative hydrogen production	45
2-5-1 Temperature	                                               46
2-5-2 pH	                                                        47
2-5-3 HRT                                                        48
2-5-4 Volatile acids	                                      49
2-5-5 Ammonia	                                               49
2-5-6 Hydrogen/ carbon dioxide partial pressure	           50
2-5-7 Iron	                                                52
2-6 Anaerobic hydrogen fermentors	                              53
2-6-1 Completely stirred tank reactors	                     53
2-6-2 UASB	                                                54
2-6-3 Membrane bioreactors	                                      55
Chapter 3 Material and Methods	58
3-1 Batch experiments for hydrogen production potential	58
3-1-1 Serum bottles	58
3-1-2 Nutrients	58
3-1-3 Substrates	59
3-1-4 Seed sludge	59
3-2 Batch experiments for biohydrogen potential with chloroform	59
3-2-1 Seed sludge	59
3-2-2 Nutrients and chemicals	60
3-2-3 Procedures	60
3-3 Effects of acetate or alumina on anaerobic hydrogen fermentation	61
3-3-1 Batch experiments of acetate at neutral pH	61
3-3-2 Batch experiments of acetate at different pH	62
3-3-3 Batch experiments of alumina	62
3-4 Effects of temperature on fermentative hydrogen production	63
3-4-1 Seed sludge	63
3-4-2 Batch experiments	63
3-4-3 Anaerobic hydrogen fermentors	64
3-5 Anaerobic hydrogen fermentors installed with a pervaporation membrane	65
3-5-1 Membrane modules	65
3-5-2 Anaerobic hydrogen fermentors	66
3-5-3 Seed sludge	67
3-5-4 Hollow fiber module served as a fermentor	68
3-5-5 Determination of mass transfer coefficients	68
3-6 Anaerobic hydrogen fermentors installed with a microfiltration membrane 69
3-6-1 Anaerobic hydrogen fermentors	69
3-6-2 Seed sludge	71
3-7 Eelectrodialysis cells to separate fermented products	71
3-7-1 ED cell	71
3-7-2 Synthetic liquid	71
3-7-3 Fermented liquid	72
3-8 Analytical methods	72
3-8-1 Analysis of gas composition	72
3-8-2 Volatile acids	72
3-8-3 Others	72
Chapter 4 Biokinetics of anaerobic hydrogen production of glucose and peptone 73
4-1 Introduction	73
4-2 Biological hydrogen potential of glucose	75
4-2-1 Hydrogen production and products from the BHP test using glucose with preheated sludge seed	75
4-2-2 Hydrogen production and products from the BHP tests using glucose with enhanced sludge seed	80
4-2-3 Hydrogen-producing rate and hydrogen yield	82
4-3 Biohydrogen production potential of peptone	84
4-4 Remarks	86
Chapter 5 Factors affecting anaerobic hydrogen fermentation	87
5-1 Introduction	88
5-2 Effects of acetate on anaerobic hydrogen fermentation	90
5-2-1 Effects of acetate on hydrogen fermentatin of peptone	90
5-2-2 Effects of acetate in different pH on hydrogen fermentatin of peptone 94
5-2-3 Effects of γ-alumina on on hydrogen fermentatin of peptone	98
5-3 Effects of temperature on anaerobic hydrogen fermentation	102
5-3-1 A comparison between batch thermophilic and mesophilic fermentation	102
5-3-2 Comparisons between continuous TAHF and MAHF bioreactors	104
5-3-3 Comparison of fermented products	106
5-4 Remarks	109
Chapter 6 Start-up of anaerobic hydrogen fermentors: chemical inhibition	110
6-1 Intruduction	111
6-2 Batch incubation with chloroform inhibition	114
6-2-1 First batch incubation with chloroform inhibition	114
6-2-2 Second batch incubation with chloroform inhibition	116
6-2-3 Third batch incubation without chloroform	117
6-2-4 Microscopy of morphology	118
6-3 Start-up of anaerobic hydrogen fermentors	121
6-3-1 Semi-batch feeding	121
6-3-2 Continuous feeding	123
6-4 Hydrogen yield	124
6-5 Remarks	127
Chapter 7 Anaerobic hydrogen fermentors installed with a silicone rubber membrane	128
7-1 Introduction	129
7-1-1 Transfer models of nonporous membrane (gas phase)	131
7-1-2 Transfer models of nonporous membrane (multiphase)	133
7-2 Mass transfer coefficients of H2 and CO2 at the silicone rubber membrane 135
7-3 Batch fermentation	138
7-4 Hollow fiber module served as a fermentor	141
7-5 Hydrogen yield	144
7-6 Hydrogen partial pressure in bioreactor	145
7-7 Remarks	146
Chapter 8 Anaerobic hydrogen Fermentors Installed with a microfiltration membrane	147
8-1 Introduction	148
8-2 Accumulation of biomass in the CSTR and MBR fermentors	149
8-3 Hydrogen-producing activity in the CSTR and MBR fermentors	157
8-4 MBR fermentor with periodically withdrawing surplus sludge	152
8-5 MBR fermentor without withdrawing surplus sludge	155
8-6 Hydrogen production of the MBR fermentor	156
8-7 Products of fermentation	158
8-8 F/M ratio and H2-production	159
8-9 Microscopy	161
8-10 Remarks	162
Chapter 9 Feasibility of separating fermented products by electrodialysis process	163
9-1 Introduction	164
9-2 Effects of HRT factor	166
9-3 Effects of voltage factor	168
9-4 Remarks	171
Chapter 10 Conclusions and Future Perspectives	172
10-1 Conclusions and suggestions	172
10-2 Future perspectives	175
References	176
Appendex 1 Presentation	191
Appendex 2 Vita	193



 
List of Figures

Figure 1-1 Outlinesof this dissertation	4
Figure 1-2 Structure of this dissertation	5
Figure 2-1 Proposal of total hydrogen production by a two-stage bioreactor	7
Figure 2-2 Energetic views of biohydrogen production	8
Figure 2-3 Anaerobic transformation of substrates	14
Figure 2-4 Fermentation of glucose by C. acetobutylicum	21
Figure 2-5 Principal routes of amino acid fermentation	25
Figure 2-6 Proposed mechanism of hydrogen production with the peptone fermentation	26
Figure 2-7 Phylogenetic affiliation of isolated strains was constructed by neighbor-joining method based on 16S rDNA	34
Figure 2-8 Hydrogen production from the batch bioreactor fed with 1 gram of glucose	38
Figure 2-9 Time courses of hydrogen production and consumption on fermenting 1 g of peptone	39
Figure 3-1 Configuration of anaerobic hydrogen fermentation bioreactors	64
Figure 3-2 Diagram of a hydrogen fermentation reactor installed with the hollow fiber membrane	68
Figure 3-3 Configuration of a one-through and continuous-stirred tank reactor (CSTR) fermentor without recovery of biomass	70
Figure 3-4 Configuration of thermophilic anaerobic hydrogen fermentor coupled with a miccrofiltration membrane	70
Figure 3-5 Configure of the ED cell	71
Figure 4-1 Cumulative hydrogen production obtained from the BHP tests using	76
Figure 4-2 Formate generated in the BHP tests seeded with preheated sludge	77
Figure 4-3 Acetate generated in the BHP tests seeded with preheated sludge	78
Figure 4-4 Propionate generated in the BHP tests seeded with preheated sludge	78
Figure 4-5 Butyrate generated in the BHP tests seeded with preheated sludge	79
Figure 4-6 Ethanol generated in the BHP tests seeded with preheated sludge	79
Figure 4-7 Composition of volatile acids produced in the BHP test using glucose with preheated sludge seed	80
Figure 4-8 Composition of volatile acids produced in the BHP test using glucose with enhanced sludge seed	81
Figure 4-9 Relationship of initial hydrogen-producing rate with initial concentration of glucose	83
Figure 4-10 Relationship of initial hydrogen yield with the ratios of So/Xo (The parameters adopted only when the consumption of glucose is over 90% in the BHP tests)	83
Figure 4-11 Cumulative hydrogen production obtained from the BHP tests using peptone with enhanced sludge seed	85
Figure 5-1 Effects of various acetate concentrations on cumulative H2 production from batch fermentation of peptone	91
Figure 5-2 Variation of hydrogen-producing rate obtained from hydrogen fermentation of peptone with concentration of extra–added acetate	92
Figure 5-3 Variation of hydrogen yield obtained from hydrogen fermentation of peptone with concentration of extra-added acetate	93
Figure 5-4 Variation of lag phase time obtained from hydrogen fermentation of peptone with concentration of extra-added acetate	93
Figure 5-5 Variation of hydrogen-producing rate obtained from hydrogen fermentation of peptone with extra-added 4,000 mg/L acetate under different initial pH	96
Figure 5-6 Variation of hydrogen yield obtained from hydrogen fermentation of peptone with extra-added 4,000 mg/L acetate under different initial pH	97
Figure 5-7 Variation of lag phase time obtained from hydrogen fermentation of peptone with extra-added 4,000 mg/L acetate under different initial pH	97
Figure 5-8 The evolution of hydrogen is enhanced via alumina added at the end of hydrogen-producing stage during the batch incubation with peptone culture	100
Figure 5-9 Effects of alumina on evolution of hydrogen in batch fermentation of peptone	100
Figure 5-10 Effects of alumina on evolution of hydrogen in batch fermentation of glucose	101
Figure 5-11 Effects of alumina on hydrogen evolution rate in batch fermentation of peptone or glucose	101
Figure 5-12 H2-producing rates of the TAHF and MAHF bioreactors	105
Figure 5-13 CO2-producing rates of the TAHF and MAHF bioreactors	105
Figure 5-14 Products obtained from fermenting one gram of substrate in the TAHF and MAHF bioreactors at 20 g COD/g VSS-day	107
Figure 5-15 Electron flow of converting one gram of substrate as COD in the TAHF and MAHF bioreactors at 20 g COD/g VSS-day	108
Figure 6-1 Time variation of biogas production from the serum bottles with addition of chloroform in the first batch incubation	115
Figure 6-2 Time variation of biogas production from the serum bottles with addition of chloroform in the second batch incubation	116
Figure 6-3 Time variation of H2 biogas production from the serum bottles without addition of chloroform in the third batch incubation (no methane obtained)	118
Figure 6-4 Morphology of raw granular sludge with filamentous methanogens	119
Figure 6-5 Morphology of first batch sludge with filamentous and rod microorganisms	120
Figure 6-6 Morphology of second batch growth with round-head rod microorganisms	120
Figure 6-7 Morphology of third batch growth with rod microorganisms	121
Figure 6-8 Cumulative gas production in fermenting glucose with semi-batch feeding	122
Figure 6-9 Evolution rates of H2 and CO2 in semi-batch feeding	123
Figure 6-10 Gas composition observed from the semi-batch bioreactors with thermal boiling and chloroform-pretreated sludge from batch incubation for fermenting glucose	124
Figure 7-1 Mass transfer model for gas permeation membrane	131
Figure 7-2 Mass transfer model of multiphase for the pervaporation membrane	134
Figure 7-3 The mass transfer coefficients (Kp) of H2 and CO2 for the hollow fiber module	137
Figure 7-4 Cumulative production of CO2 across the hollow fiber module and into the headspace of the reactor in batch fermentation	140
Figure 7-5 Cumulative production of H2 across the hollow fiber module and into the headspace of the reactor in batch fermentation	140
Figure 7-6 Cumulative hydrogen production of fermenting glucose in the hollow fiber module as a fermentor	141
Figure7-7 The evolution rate of fermenting glucose in the membrane module as a fermentor	143
Figure 7-8 Ratio of the evolution rate of H2 to that of CO2 in the batch fermentation of glucose	143
Figure7-9 Comparisons between the hydrogen evolution rates of the fermentors with or without silicone rubber membrane	144
Figure 8-1 Variation of biomass accumulated in the CSTR and MBR fermentors	150
Figure 8-2 Ratios of H2 to CO2 in the biogas from the CSTR and MBR fermentors	150
Figure 8-3 Concentration of biomass in the MBR fermemtor	153
Figure 8-4 Hydrogen-producing rates in the MBR fermentor	154
Figure 8-5 Biomass and specific gas-producing rate in thermophilic anaerobic hydrogen fermemtor coupled with a membrane	156
Figure 8-6 Variation of liquid products and F/M ratio from the MBR fermentor	159
Figure 8-7 Relationship of specific H2-producing rates and F/M from the thermophilic MBR fermentor	160
Figure 8-8 Relationship of hydrogen yield and F/M from the thermophilic MBR fermentor	160
Figure 8-9 Morphology of the CSTR fermentor at 6 h HRT	161
Figure 8-10 Morphology of the MBR fermentor at 6 h HRT and near 2 day SRT	161
Figure 9-1 Whole picture of biohydrogen processes	164
Figure 9-2 Function of electordialysis	166
Figure 9-3 Separation of ammonium and acetate in effluent dilution at different HRT ( E=10 Volt, Distance=6 cm)	167
Figure 9-4 Conductivity of effluent dilution at different HRT ( E=10 Volt, Distance=6 cm)	167
Figure 9-5 Separation of ammonium and acetate in effluent dilution at different voltage (HRT=9 min, Distance=0.8)	169
Figure 9-6 Residual concentration of acetic acid in dilution depending on its pH	169
Figure 9-7 Conductivity in effluent dilution at different voltage (HRT=9 min Distance=0.8)	170
Figure 10-1 Future perspectives on application of membrane installed with biohydrogen fermentor 	175
	

 
List of Tables 

Table 2-1 Advantages and disadvantages of different biological processes for hydrogen production	9
Table 2-2 Microorganisms used for photosynthetic hydrogen generation	11
Table 2-3 The influences of different operational conditions on photosynthetic hydrogen production	12
Table 2-4 Reductive potential of enzymatic reaction in biological systems	17
Table 2-5 Free energy of hydrogen production from organic compounds	18
Table 2-6 Bacterial fermentation with carbohydrates as substrates	22
Table 2-7 Microorganisms used for hydrogen generation	28
Table 2-8 Characteristics of hydrogen production of Enterobacter	32
Table 2-9 Phylogenetic affiliations of the 16S rDNA clone library of the biohydrogen reactor with sucrose as influent	35
Table 2-10 Phylogenetic affiliations of the 16S rDNA clone library of the biohydrogen reactor with wheat husk as influent	36
Table 2-11 Kinetic parameters of hydrogen production with Gompertz equation	38
Table 2-12 Biokinetics of substrate conversion in mesophilic anaerobic digestion	42
Table 2-13 Estimated yield coefficients and substrate-based yields for products resulting from H2 fermentation	42
Table 2-14 Hydrogen yields reported in the literature	43
Table 2-15 Effects of low pH condition on anaerobic hydrogen-producing bacteria	47
Table.3-1 Composition of the nutrients for batch hydrogen production tests	58
Table 3-2 The first batch incubation with chloroform inhibition	60
Table 3-3 The second batch incubation with chloroform inhibition	61
Table 3-4 The third batch incubation with chloroform inhibition	61
Table 3-5 Components for preparing the culture medium	65
Table 3-6 Specific of the membrane module made of silicone rubber.	66
Table 4-1 Parameters and products of the BHP tests using glucose with preheated sludge seed	77
Table 4-2 Parameters and products of the BHP tests using glucose with enhanced sludge seed	81
Table 4-3 Parameters and products of the BHP tests using peptone with enhanced sludge seed	85
Table 5-1 Parameters obtained from the trials with various acetate concentrations added in hydrogen fermentation of peptone	92
Table 5-2 Parameters from various pH affecting on hydrogen fermentation of peptone with extra-added 4,000 mg/L acetate	95
Table 5-3 Volatile acids and ammonium from fermentation of glucose or peptone	102
Table 5-4 Parameters of the Gompertz equation in TAHF and MAHF of 160 mg peptone or glucose	103
Table 5-5 Gas-producing rates of TAHF and MAHF bioreactors	106
Table 5-6 Products obtained by fermenting one gram of substrate in the TAHF and MAHF bioreactors at 20 g COD/g VSS-day	107
Table 5-7 Electron flow of converting one gram of substrate as COD in the TAHF and MAHF bioreactors at 20 g COD/g VSS-day	108
Table 6-1 Hydrogen producing by chloroform-pretreated sludge in the first batch incubation	115
Table 6-2 Hydrogen producing by chloroform-pretreated sludge in the second batch incubation	117
Table 6-3 Hydrogen producing by chloroform-pretreated sludge in the third batch incubation	117
Table 6-4 Comparison of hydrogen yield in terms of one gram of substrate applied	125
Table 6-5 Production of hydrogen and volatile fatty acids obtained from thermal boiling and chloroform -pretreated sludge by fermentation of peptone and glucose	126
Table7-1 Gas permeability of silicone rubber	133
Table 7-2 The flux and gas composition in the collection bottles connected to the membrane and the fermentor headspace	136
Table 7-3 Mass transfer coefficients of the hollow fibers with silicone rubber membrane	136
Table7-4 Gas evolution rates in logarithmic phase in batch fermentation of glucose	139
Table 8-1 Hydrogen production of the CSTR and MBR fermentors	151
Table 8-2 Parameters of thermophilic hydrogen fermentation after a membrane installed with the fermentor	157
Table 9-1 Preliminary test using batch ED on the effluent from the biohydrogen fermentor (E= 18V, Distance =6 cm)	170
                                    

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