Continuous biogenic hydrogen production from dilute acid pretreated algal hydrolysate using hybrid immobilized mixed consortia

This study investigated the bioconversion of dilute acid (2% H₂SO₄) pretreated red algae (Gelidium amansii) hydrolysate into H₂ by anaerobic fermentation in a continuous stirred tank reactor under mesophilic conditions using hybrid immobilized cells as microbial catalyst. Two different hydraulic retention times (HRT) of 24 h and 16 h with a feed concentration of 15 g/L hexose equivalent have been investigated over 85 days of operation to evaluate H₂ production performance and stability of the continuous system. The highest hydrogen production rate (HPR) and hydrogen yield (HY) of 2.7 L/L/d and 1.3 mol/mol substrate hexose_added was achieved at 24 h HRT, while further operation at 16 h HRT led to a significant drop in the hydrogen production with a HPR and HY values of 1.8 L/L/d and 0.7 mol/mol substrate hexose_added, respectively. The bacterial community analysis characterized by 454 pyrosequencing revealed that the changes in HRT significantly influence the composition of the dominant microflora. At longer HRT (24 h), the phyla Firmicutes was abundant over 98%, whereas at shorter HRT (16 h), Proteobacteria being the dominant populations with 84%. These outcomes suggested that controlling appropriate HRT is prerequisite for efficient hydrogen production.     

Enhanced biohydrogen production from tofu residue by acid/base pretreatment and sewage sludge addition

Anaerobic dark fermentation is considered a promising technology for clean energy production and waste reduction. In the present work, tofu residue and sewage sludge were utilized as substrates for fermentative hydrogen production. To increase the biodegradability, tofu residue was pretreated for 30 min in the presence of HCl and NaOH at various concentrations (0, 0.5, 1.0, and 2.0%), and then fermented by a thermophilic (60 °C) mixed culture. The solubility (SCOD/TCOD ratio) of the tofu residue increased from 4 to 30–40% after pretreatment, and the increased soluble constituents were mainly protein rather than carbohydrate compounds. However, in spite of a slight increase of carbohydrate solubility, the H₂ production performance was significantly enhanced by pretreatment, owing to the degradability of thermophilic cultures used on insoluble tofu residues. The limited H₂ yield of 0.30 mol H₂/mol hexose_added achieved in the raw tofu residue was increased 1.6- to 4-fold with the highest H₂ yield of 1.25 mol H₂/mol hexose_added at 1.0% HCl concentration. Carbohydrate degradation and the H₂ production rate also increased from 39 to 50–65% and 27 to 50–120 mL H₂/L/h, respectively. The role of pretreatment was not only to increase the biodegradability but also to suppress the activity of indigenous non H₂-producers such as lactic acid bacteria and propionic acid bacteria. When sewage sludge was added to acid pretreated (1.0% HCl) tofu residue as a co-substrate, the H₂ yield and H₂ production rate increased to 1.48 mol H₂/mol hexose_added and 161 mL H₂/L/h, respectively, which was attributed to the abundant minerals, vitamins, and metals contained in sewage sludge.     

Non-sterile bio-hydrogen fermentation from food waste in a continuous stirred tank reactor (CSTR): Performance and population analysis

Bio-hydrogen production from food waste by anaerobic mixed cultures was conducted in a continuous stirred tank reactor (CSTR). The hydraulic retention time (HRT) was optimized in order to maximize hydrogen yield (HY) and hydrogen production rate (HPR). The maximum hydrogen content (38.6%), HPR (379 mL H₂/L. d) and HY (261 mL H₂/g-VS_added) were achieved at the optimum HRT of 60 h. The major soluble metabolite products were butyric and acetic acids which indicated a butyrate-acetate type fermentation. Operation of CSTR at HRT 60 h could select hydrogen producing bacteria and eliminate lactic acid bacteria and acetogenic bacteria. The microbial community analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) revealed that the predominant hydrogen producer was Clostridium sp.     

Continuous fermentative hydrogen production from coffee drink manufacturing wastewater by applying UASB reactor

The feasibility of continuous H₂ production from coffee drink manufacturing wastewater (CDMW) was tested in two different types of reactors: a completely-stirred tank reactor (CSTR) and an up-flow anaerobic sludge blanket reactor (UASBr). While the performance in CSTR was limited, it was significantly enhanced in UASBr. The maximum H₂ yield of 1.29 mol H₂/mol hexose_added was achieved at HRT of 6 h in UASBr operation. Non-hydrogenic, lactic acid was the dominant in CSTR, while butyric and caproic acids in UASBr. As caproic acid is generated by consuming acetic and butyric acids, all of which are related to H₂ production, the presence of caproic acid in the broth also indicates H₂ production, yielding 1.33 mol H₂/glucose. It was speculated that the enhanced performance in UASBr was attributed to the high concentration of biomass over 60,000 mg VSS/L in the blanket zone, which provided insufficient substrate for indigenous lactic acid bacteria (LAB) to survive. The abundance of LAB in CDMW was confirmed by natural fermentation of CDMW. That is without the addition of external inoculum, CDMW was mainly fermented into lactic acid under mesophilic condition. For the first time ever, H₂ producing granules (HPG) with diameters of 2.1 mm were successfully formed by using actual waste as a substrate.     

Continuous biohydrogen production using cheese whey: Improving the hydrogen production rate

Due to the renewed interest in finding sustainable fuels or energy carriers, biohydrogen (Bio-H₂) from biomass is a promising alternative. Fermentative Bio-H₂ production was studied in a continuous stirred tank reactor (CSTR) operated during 65.6 d with cheese whey (CW) as substrate. Three hydraulic retention times (HRTs) were tested (10, 6 and 4 h) and the highest volumetric hydrogen production rate (VHPR) was attained with HRT of 6 h. Therefore, four organic loading rates (OLRs) at a fixed HRT of 6 h were tested thereafter, being: 92.4, 115.5, 138.6 and 184.4 g lactose/L/d. The highest VHPR (46.61 mmol H₂/L/h) and hydrogen molar yield (HMY) of 2.8 mol H₂/mol lactose were found at an OLR of 138.6 g lactose/L/d; a sharp fall in VHPR occurred at an OLR of 184.4 g lactose/L/d. Butyric, propionic and acetic acids were the main soluble metabolites found, with butyric-to-acetic ratios ranging from 1.0 to 2.4. Bacterial community was identified by partial sequence analysis of the 16S rRNA and polymerase chain reaction–denaturing gradient gel electrophoresis (PCR–DGGE). The results showed that at HRT of 10 h and 6 h were dominated by the Clostridium genus. The VHPR attained in this study is the highest reported value for a CSTR system using CW as substrate with anaerobic sludge as inoculum and represents a 33-fold increase compared to a previous study. Thus, it was demonstrated that continuous fermentative Bio-H₂ production from CW can be significantly enhanced by an appropriate selection of parameters such as HRT and OLR. Enhancements in VHPR are significant because it is a critical parameter to determine the full-scale practical application of fermentation technologies that will be used for sustainable and clean energy generation.     

A simple method to reduce the start-up period in a H2-producing UASB reactor

Introduction of an up-flow anaerobic sludge blanket (UASB) reactor apparatus to fermentative hydrogen production (FHP) enormously improves H₂ production performance. However, the long start-up period required to form H₂-producing granules (HPG) remains as a major obstacle. In the present work, a completely-stirred tank reactor (CSTR) was operated for 7 days, and the mixed liquor in the CSTR was transferred to a UASB reactor (UASBr (I)) as a seeding source. Coffee drink manufacturing wastewater (CDMW) was used as a feedstock, constituting the first attempt to form HPG from actual industrial wastewater. The strategy employed here was found to be more effective in developing HPG than directly starting from the UASB reactor (UASBr (II)), which is attributed to substantially higher active mass transfer in the CSTR. The average size of particles in the UASBr (II) blanket zone after 50 days of operation corresponded with that in the CSTR after only 7 days of operation. The drastic decrease of extracellular polymeric substance (EPS) protein concentration in the CSTR operation also indicates efficient removal of non-active biomass, the presence of which could adversely affect HPG formation. UASBr (I) showed a stable H₂ yield and H₂ production rate of 1.78 mol H₂/mol hexose_added and 2.76 L H₂/L/h, respectively, and HPG with an average size of 1.9 mm were developed after 45 days. It appears that the abundant presence of divalent ions, especially calcium ions, contained in the CDMW facilitated HPG formation.     

Effect of storage time and temperature on hydrogen fermentation of food waste

Practically, before being fed to the treatment plant, food waste (FW) is stored for up to a week in a storage tank under ambient temperature condition, which would have an impact on the bioenergy yield. In the present work, FW was stored at different temperatures (5 °C, 20 °C, and 35 °C) for 0 d, 1 d, and 2 d, and it was used as a feedstock for mesophilic H₂ fermentation. H₂ production curves were divided by three groups, finally attaining 1.7–1.8 mol H₂/mol hexose_added, 1.4–1.5 mol H₂/mol hexose_added, and 1.2 mol H₂/mol hexose_added, achieved from the (fresh, FW stored at 5 °C), (FW stored at 20 °C, and 35 °C for 1 d), and (FW stored at 35 °C for 2 d), respectively. The different performance was attributed to the growth of indigenous lactic acid bacteria such as Lactobacillus and Weissella during storage under high temperature condition. In addition, it was found that the activity of homoacetogenic reaction (R17, 4H₂ + CO₂ → Acetate) calculated by establishing metabolic flux balance was different depending on the H₂ production performance. The flux of R17 ranged 0.03–0.06 under low H₂ yield achieved conditions, while it increased to 0.10–0.17 those showing low H₂ yields.     

Enhancing the performance of dark fermentative hydrogen production using a reduced pressure fermentation strategy

The partial pressure of hydrogen is an extremely important factor for hydrogen generation. This study investigated the effect of reduced pressure (via vacuum) on hydrogen production in a CSTR reactor. The results show that the reduced pressure condition is more effective in enhancing H₂ production at lower HRT (e.g., 8–4 h) than at higher HRT (e.g., 12 h). The optimal hydrogen yield and overall hydrogen production efficiency occurred at a HRT of 6 h with a value of 4.50 mol H₂/mol sucrose and 56.2%, respectively. Meanwhile, at HRT 6 h the hydrogen production rate was 0.937 mol/L/d. In addition, the HPR could be further improved to 1.196 mol/L/d when the HRT was shortened to 4 h, obtaining a 37–271% increase in HPR when compared with that described in the relevant reports. For all experiments, butyrate and acetate were the two primary soluble metabolites, accounting for 85–99% of total soluble microbial products. Predominant production of acetate and butyrate demonstrates the efficient H₂ fermentation with reduced pressure processes.     

Two-stage UASB reactor converting coffee drink manufacturing wastewater to hydrogen and methane

In this study, the feasibility of a continuous two-stage up-flow anaerobic sludge blanket (UASB) reactor system, consisted of thermophilic (55 °C) dark fermentative H₂ production and mesophilic (35 °C) CH₄ production from coffee drink manufacturing wastewater (CDMW), was tested. A recently proposed operational strategy was used to overcome a major drawback of the long start-up period of the UASB reactor. Firstly, a completely stirred tank reactor (CSTR) was operated for 8 days to prepare seeding. The seed was then directly transferred to the UASB reactor. Microbial aggregation took place in the initial period, and the floc size was gradually increased over time. In UASB reactor, the maximum H₂ yield of 2.57 mol H₂/mol hexose_added and a stable H₂ production rate of 4.24 L H₂/L/h were observed at a hydraulic retention time (HRT) of 6 h and substrate concentration of 20 g Carbo. COD/L. In this novel method using CDMW, thermophilic H₂-producing granules with an average particle size of 1.3 mm was successfully developed after 100 days. The more bioenergy recovery was attempted in a post-treatment process using a mesophilic UASB reactor for CH₄ production from the H₂ fermented effluent. The maximum CH₄ yield of 325 mL of CH₄/g COD was achieved with removing 93% of the COD at an organic loading rate of 3.5 g COD/L/d. The developed two-stage UASB reactor system achieved biogas conversion by 88.2% (H₂ 15.2% and CH₄ 73%) and COD removal by 98%.     

Experience of a pilot-scale hydrogen-producing anaerobic sequencing batch reactor (ASBR) treating food waste

A pilot-scale H₂-producing anaerobic sequencing batch reactor (ASBR) treating food waste was operated. During the operation, the carbon/nitrogen (C/N) ratio was adjusted from 10 to 30 by changing the composition of the food waste. When the C/N ratio was lower than 20, the H₂ yield was maintained at around 0.5 mol H₂/mol hexose_added, accounting for 2.3% of energy conversion efficiency contained in food waste to H₂, but it gradually dropped at higher C/N ratios. The low performance was accompanied by increased production of lactate, propionate, and valerate. In order to recover the performance, alkaline shock (pH 12.5 for 1 day) was imposed on the entire mixed liquor in the fermenter. This alkaline shock method was so effective that the H₂ yield significantly increased to over 0.9 mol H₂/mol hexose_added, and was then stabilized at 0.69 mol H₂/mol hexose_added. In addition, the settling characteristics of H₂-producing ASBR, which was separated into three layers, were investigated. Ribonucleic acid (RNA) as well as volatile suspended solid concentrations of each layer were measured to suggest how to enhance the H₂ production in ASBR operation.     

The effect of hydraulic retention time on thermophilic dark fermentative biohydrogen production in the continuously operated packed bed bioreactor

The main objective of the study is to investigate the effect of hydraulic retention times on continuous dark fermentative biohydrogen production in an up-flow packed bed reactor (UPBR) containing a novel microorganism immobilization material namely polyester fiber beads. The hydrogen producing dark fermentative microorganisms were obtained by heat-pretreatment of anaerobic sludge from the acidogenic phase of an anaerobic wastewater treatment plant. Glucose was the sole carbon source and the initial concentration was 15 ± 1 g/L throughout the continuous feeding. UPBR was operated under the thermophilic condition at T = 48 ± 2 °C and at varying HRTs between 2 h and 6 h. The hydrogen productivity of continuously operated UPBR increased with increasing HRT. Hydrogen production volume varied between 4331 and 6624 ml/d, volumetric hydrogen production rates (VHPR) were obtained as 3.09–4.73 L H₂/L day, and hydrogen production yields (HY) were 0.49 mol/mol glucose-0.89 mol/mol glucose depending on HRT. Maximum daily hydrogen volume (6624 ml/d), the yield (0.89 mol/mol glucose) and VHPR (4.73 L H₂/L day) were obtained at HRT = 6 h. The production rate and the yield decreased with increasing organic loading rate due to substrate inhibition.     

Direct fermentation of Laminaria japonica for biohydrogen production by anaerobic mixed cultures

A few studies have been made on fermentative hydrogen production from marine algae, despite of their advantages compared with other biomass substrates. In this study, fermentative hydrogen production from Laminaria japonica (one brown algae species) was investigated under mesophilic condition (35 ± 1 °C) without any pretreatment method. A feasibility test was first conducted through a series of batch cultivations, and 0.92 mol H₂/mol hexose_added, or 71.4 ml H₂/g TS of hydrogen yield was achieved at a substrate concentration of 20 g COD/L (based on carbohydrate), initial pH of 7.5, and cultivation pH of 5.5. Continuous operation for a period of 80 days was then carried out using anaerobic sequencing batch reactor (ASBR) with a hydraulic retention time (HRT) of 6 days. After operation for approximately 30 days, a stable hydrogen yield of 0.79 ± 0.03 mol H₂/mol hexose_added was obtained. To optimize bioenergy recovery from L. japonica, an up-flow anaerobic sludge blanket reactor (UASBr) was applied to treat hydrogen fermentation effluent (HFE) for methane production. A maximum methane yield of 309 ± 12 ml CH₄/g COD was achieved during the 90 days operation period, where the organic loading rate (OLR) was 3.5 g COD/L/d.     

Continuous photo-fermentative hydrogen production from lactate and lactate-rich acidified food waste

In the present work, a continuous photo-fermentative hydrogen (H₂) production from lactate was attempted at various hydraulic retention times (HRTs) (48–120 h). Electron balance was made at each operating conditions to elucidate different performances. At 120 h HRT, H₂ production was negligible, while 42% and 52% of substrate electrons diverted towards cell growth and soluble microbial products (SMPs), respectively. After changing HRT to 96 h, H₂ yield jumped to 2.3 mol-H₂/mol-lactate_added with less SMPs production and minimal cell growth. The highest H₂ production rate of 260 mL-H₂/L-fermenter/d was also achieved at 96 h HRT. When HRT was further shortened to 72 and 48 h, H₂ yield dropped to 1.4 and 0.2 mol-H₂/mol-lactate_added, respectively. While almost all of the lactate was degraded at <72 h HRT, only 65% of the lactate was consumed at 48 h HRT. From 200th day, the feedstock was changed to lactate-rich acidified food waste, which was obtained via one-day fermentation of food waste and subsequent centrifugation. At 2 g chemical oxygen demand/L, substrate conversion efficiency reached 35%, which was slightly lower than that of feeding pure lactate. SMPs were found to be mainly consisted of low molecular weight compounds (<500 Da), and the majority of organic matters were aromatic proteins at 120 h HRT and it was shifted to humic-like region in 96 h HRT.     

Influence of Cu concentration on the biohydrogen production of continuous stirred tank reactor

In this paper, the effect of hydraulic retention time (HRT, 16 h–4 h) on fermentative hydrogen production by mixed cultures was firstly investigated in a sucrose-fed anaerobic continuous stirred tank reactor (CSTR) at 35 °C and initial pH 8.79. After stable operations at HRT of 16–6 h, the bioreactor became unstable when the HRT was lowered to 4 h. The maximum hydrogen yield reached 3.28 mol H₂/mol-Sucrose at HRT 4 h. Supplementation of Cu²⁺ at HRT 4 h improved the operation stability through enhancement of substrate degradation efficiency. The effect of Cu²⁺ concentration ranging from 1.28 to 102.4 mg/L on fermentative hydrogen production was studied. The results showed that Cu²⁺ was able to enhance the hydrogen production yield with increasing Cu²⁺ concentration from 1.28 to 6.4 mg/L. The maximum hydrogen yield of 3.31 mol H₂/mol-Sucrose and the maximum hydrogen production rate of 14.44 L H₂/Day/L-Reactor were obtained at 6.4 mg/L Cu²⁺ and HRT 4 h Cu²⁺ at much higher concentration could inhibit the hydrogen production, but it could increase substrate degradation efficiency (12.8 and 25.6 mg/L Cu²⁺). The concentration of Cu²⁺ had effect on the distribution of soluble metabolite.     

Thermophilic fermentative hydrogen production from various carbon sources by anaerobic mixed cultures

In the present work, various carbon sources, xylose, glucose, galactose, sucrose, cellobiose, and starch were tested for thermophilic (60 °C) fermentative hydrogen production (FHP) by using the anaerobic mixed culture. An inoculum was obtained from a continuously-stirred tank reactor (CSTR) operated at pH 5.5 and HRT 12 h, and fed with tofu processing waste. The dominant species in the CSTR were found to be Thermoanaerobacterium thermosaccharolyticum and Clostridium thermosaccharolyticum, which are well known thermophilic H₂-producers in anaerobic-state, and have the ability to utilize a wide range of carbohydrates. When initial pH was adjusted to 6.8 ± 0.1 but not controlled during fermentation, vigorous pH drop began within 5 h, and finally reached 4.0–4.5 in all carbon sources. Although over 90% of substrate removal was achieved for all carbon sources except cellobiose (71.7%), the fermentation performances were profoundly different with each other. Glucose, galactose, and sucrose exhibited relatively higher H₂ yields whereas lower H₂ yields were observed for xylose, cellobiose, and starch. On the other hand, when pH was controlled (pH ≥ 5.5), the fermentation performance was enhanced in all carbon sources but to a different extent. A substantial increase in H₂ production was observed for cellobiose, a 1.9-fold increase of H₂ yield along with a substrate removal increase to 93.8%, but a negligible increase for xylose. H₂ production capabilities of all carbon sources tested were as follows: sucrose > galactose > glucose > cellobiose > starch > xylose. The maximum H₂ yield of 3.17 mol H₂/mol hexose_added achieved from sucrose is equivalent to a 26.5% conversion of energy content in sucrose to H₂. Acetic and butyric acids were the main liquid-state metabolites of all carbon sources while lactic acid was detected only in cellobiose, starch and xylose exhibiting relatively lower H₂ yields.     

Feasibility of biohydrogen production from Gelidium amansii

The feasibility of hydrogen production from red algae was investigated. Galactose, the main sugar monomer of red algae, was readily converted to hydrogen by dark fermentation. The maximum hydrogen production rate and yield of galactose were 2.46 L H₂/g VSS/d and 2.03 mol H₂/mol galactose_added, respectively, which were higher than those for glucose (0.914 L H₂/g VSS/d and 1.48 mol H₂/mol galactose_added). The distribution of soluble byproducts showed that H₂ production was the main pathway of galactose uptake. 5-HMF, the main byproduct of acid hydrolysis of red algae causes noncompetitive inhibition of H₂ fermentation. 1.37 g/L of 5-HMF decreased hydrogen production rate by 50% compared to the control. When red algae was hydrolyzed at 150 °C for 15 min and detoxified by activated carbon, 53.5 mL of H₂ was produced from 1 g of dry algae with a hydrogen production rate of 0.518 L H₂/g VSS/d. Red algae, cultivable on vast tracts of sea by sunlight without any nitrogen-based fertilizer, could be a suitable substrate for biohydrogen production.     

Hydrogen production using sono-biohydrogenator

Hydrogen production in a novel sonicated biological hydrogen reactor (SBHR) was investigated and compared with a continuous stirred tank reactor (CSTR). The two systems were operated at a hydraulic retention time (HRT) of 12 h and two organic loading rates (OLRs) of 21.4 and 32.1 g COD/L.d. The average hydrogen production rates per unit reactor volume for the conventional CSTR were 2.6 and 2.8 L/L.d, as compared with 4.8 and 5.6 L/L.d for SBHR, at the two OLRs, respectively. Hydrogen yields of 1.2 and 1.0 mol H₂/mol glucose were observed for the CSTR, respectively, while for the SBHR, the hydrogen yields were 2.1 and 1.9 mol H₂/mol glucose at the two OLRs, respectively. The hydrogen content in the SBHR’s headspace was higher than that in CSTR by 10% and 31% at OLRs of 21.4 and 32.1 g COD/L.d, respectively. Both glucose conversion efficiency and HAc/HBu ratio in the SBHR were higher than in the conventional CSTR at both OLRs. The biomass yield of about 0.32 g VSS/g COD observed in the CSTR and 0.23 g VSS/g COD in the SBHR substantiate the higher H₂ yield in the SBHR. DGGE analysis confirmed the specificity of the microbial hydrogen-producing culture in the SBHR, with two different hydrogen producers (Clostridium sp. and Citrobacter freundii) detected in the SBHR and not detected in the CSTR.     

Effect of organic loading rate on fermentative hydrogen production from continuous stirred tank and membrane bioreactors

The influence of organic loading rates (OLRs) on the performance of fermentative hydrogen-producing bioreactors operating in continuous stirred tank reactor (CSTR) and membrane bioreactor (MBR) modes was examined. Five OLRs were examined, ranging from 4.0 to 30 g COD L^?1 d^?1, with influent glucose concentrations ranging from 1.3 to 10 g COD L^?1. At OLRs up to 13 g COD L^?1 d^?1, all influent glucose was utilized and the H₂ yield was not significantly influenced by OLR, although the yield in the CSTR mode was significantly higher than that in the MBR mode, 1.25 versus 0.97 mol H₂ (mol Gluc. Conv.)^?1, respectively. At an OLR of 30 g COD L^?1 d^?1, both reactor modes were overloaded with respect to glucose utilization and also had significantly higher H₂ yields of 1.77 and 1.49 mol H₂ (mol Gluc. Conv.)^?1 for the CSTR and MBR modes, respectively, versus the underloaded operation. At the intermediate OLR of 22 g COD L^?1 d^?1, the H₂ yield was maximized at 1.78 mol H₂ (mol Gluc. Conv.)^?1 for both the CSTR and MBR operation. Overall H₂ production was 50% higher in the MBR mode, 0.78 versus 0.51 moles d^?1, because the CSTR mode was overloaded with respect to glucose utilization at this OLR. These results suggest that an optimum OLR that maximizes H₂ yield and H₂ production may be near the OLR that causes overload with respect to substrate utilization. Additionally, while the CSTR mode is easier to operate and provides higher H₂ yields at underloaded and overloaded OLRs, the MBR mode may be preferable when operating near the optimum OLR.     

Aspect ratio effect of bioreactor on fermentative hydrogen production with immobilized sludge

The phenomenon of bacterial wash-out frequently occurs in the traditional continuous stirred tank reactor (CSTR) systems at low hydraulic retention time (HRT). In this study, the effect of different aspect ratios, height (H) to diameter (D) of 1:1, 3:1 and 5:1, of a CSTR with immobilized anaerobic sludge on hydrogen (H₂) production were investigated. The pH, volatile suspended solids (VSS) and total solids (TS) concentrations of the seed sludge were 6.8, 33.3 and 65.1 g/L, respectively. Thermally treated sludge was immobilized by silicone gel entrapment approach. The entrapped-sludge system operated stably at a low HRT without suffering from cell wash-out. Hence, the hydrogen production rate (HPR) was enhanced by increasing organic loading rates. The immobilized sludge CSTRs were operated at 40 °C with sucrose (10, 20, 30 and 40 g COD/L) and Endo nutrient medium at different HRTs (4, 2, 1 and 0.5 h). It was found that the granule formation enhanced HPR. The maximum HPR and the H₂ yield were found to be 15.36H₂ L/h/L and 3.16 mol H₂/mol sucrose, respectively, with the H₂ content in the biogas above 44% for all tests runs.     

Natural inducement of hydrogen from food waste by temperature control

An easy and simple method of producing H₂ from food waste was devised. Although there was no inoculum addition or pretreatment, food waste was naturally decomposed and converted to H₂ when cultivated at 50-60 °C in anaerobic state. Both the highest H₂ yield of 1.79 mol H₂/mol hexose_added and a production rate of 369.1 ml H₂/L/h were observed at 50 °C. While butyrate was the main by-product of the food waste cultivated at 50 °C, lactate whose producing-reaction is non-hydrogenic was dominant at 35 °C where the worst performance was observed. The degradation efficiency of volatile solids and carbohydrate was similar to 50% and 90%, respectively, at both temperatures. Polymerase chain reaction-denaturing gradient gel electrophoresis analysis clearly revealed that the role of temperature control was the microbial selection. At high temperature, the activity of indigenous lactic acid bacteria was suppressed while H₂-producing bacteria, such as Clostridium sp., Acetanaerobacterium elongatum, and Caloramater indicus, were predominantly cultivated.