Biohydrogen production from dual digestion pretreatment of poultry slaughterhouse sludge by anaerobic self-fermentation

Poultry slaughterhouse sludge from chicken processing wastewater treatment plant was tested for their suitability as a substrate and inoculum source for fermentation hydrogen production. Dual digestion of poultry slaughterhouse sludge was employed to produce hydrogen by batch anaerobic self-fermentation without any extra-seeds. The sludge (5% TS) was dual digested by aerobic thermophilic digestion at 55 °C with the varying retention time before using as substrate in anaerobic self-fermentation. The best digestion time for enriching hydrogen-producing seeds was 48 h as it completely repressed methanogenic activity and gave the maximum hydrogen yield of 136.9 mL H₂/g TS with a hydrogen production rate of 2.56 mL H₂/L/h. The hydrogen production of treated sludge at 48 h (136.9 mL H₂/g TS) was 15 times higher than that of the raw sludge (8.83 mL H₂/g TS). With this fermentation process, tCOD value in the activated sludge could be reduced up to 30%.     

Sequential fermentation of hydrogen and methane from steam-exploded sugarcane bagasse hydrolysate

The goal of this study was to sequential fermentation of hydrogen and methane from sugarcane bagasse (SCB). Steam explosion conditions for pretreating SCB were optimum at 195 °C and 1.5 min, which yielded 36.35 g/L of total sugar and 2.35 g/L of total inhibitors. Under these conditions (all in g/L): glucose, 11.33; xylose, 24.41; arabinose, 0.61; acetic acid, 2.33; and furfural, 0.02 were obtained. The resulting hydrolysate was used to produce hydrogen by anaerobic mixed cultures. A maximum hydrogen production rate of 396.50 mL H₂/L day was achieved at an initial pH of 6 and an initial total sugar concentration of 10 g/L. The effluent from the hydrogen fermentation process was further used to produce methane. Response surface methodology with central composite design was used to obtain the suitable conditions for maximizing methane production rate (MPR). An MPR of 185.73 mL/L day was achieved at initial pH, Ni and Fe concentrations of 7.59, 3.61 mg/L and 8.44 mg/L, respectively. Total energy of 304.11 kJ/L-substrate was obtained from a sequential fermentation of hydrogen and methane. This approach will not only add value to SCB, in the form of safe and clean energy, but also provide a solution for making use of this abundant waste.     

Biohydrogen production from sugarcane bagasse hydrolysate by elephant dung: Effects of initial pH and substrate concentration

Pre-heated elephant dung was used as inoculum to produce hydrogen from sugarcane bagasse (SCB) hydrolysate. SCB was hydrolyzed by H₂SO₄ or NaOH at various concentrations (0.25-5% volume) and reaction time of 60 min at 121 °C, 1.5 kg/cm² in the autoclave. The optimal condition for the pretreatment was obtained when SCB was hydrolyzed by H₂SO₄ at 1% volume which yielded 11.28 g/L of total sugar (1.46 g glucose/L; 9.10 g xylose/L; 0.72 g arabinose/L). The maximum hydrogen yield of 0.84 mol H₂/mol total sugar and the hydrogen production rate of 109.55 mL H₂/L day were obtained at the initial pH 6.5 and initial total sugar concentration 10 g/L. Hydrogen-producing bacterium (Clostridium pasteurianum) and non hydrogen-producing bacterium (Flavobacterium sp.) were dominating species in the elephant dung and in hydrogen fermentation broth. Sporolactobacillus sp. was found to be responsible for a low hydrogen yield obtained.     

Extreme-thermophilic biohydrogen production by an anaerobic heat treated digested sewage sludge culture

Biohydrogen production process from glucose using extreme-thermophilic H₂-producing bacteria enriched from digested sewage sludge was investigated for five cycles of repeated batch experiment at 70 °C. Heat shock pretreatment was used for preparation of hydrogen-producing bacteria comparing to an untreated anaerobic digested sludge for their hydrogen production performance and responsible microbial community structures. The results showed that the heat shock pretreatment completely repressed methanogenic activity and gave the maximum hydrogen production yield of 355-488 ml H₂/g COD in the second cycle of repeated batch cultivation with more stable gas production during the cultivation when compared with control. Hydrogen production was accompanied by production of acetic acid. The average specific hydrogen in five cycles experiment ranged from 150 to 200 ml H₂/g VSS. PCR-DGGE profiling showed that the extreme-thermophilic culture predominant species were closely affiliated to Thermoanaerobacter pseudethanolicus.     

Simultaneous saccharification and fermentation of cellulose for bio-hydrogen production by anaerobic mixed cultures in elephant dung

The objective of this study was to optimize the culture conditions for simultaneous saccharification and fermentation (SSF) of cellulose for bio-hydrogen production by anaerobic mixed cultures in elephant dung under thermophilic temperature. Carboxymethyl cellulose (CMC) was used as the model substrate. The investigated parameters included initial pH, temperature and substrate concentration. The experimental results showed that maximum hydrogen yield (HY) and hydrogen production rate (HPR) of 7.22 ± 0.62 mmol H₂/g CMC_added and 73.4 ± 3.8 mL H₂/L h, respectively, were achieved at an initial pH of 7.0, temperature of 55 °C and CMC concentration of 0.25 g/L. The optimum conditions were then used to produce hydrogen from the cellulose fraction of sugarcane bagasse (SCB) at a concentration of 0.40 g/L (equivalent to 0.25 g/L cellulose) in which an HY of 7.10 ± 3.22 mmol H₂/g cellulose_added. The pre-dominant hydrogen producers analyzed by polymerase chain reaction-denaturing gel gradient electrophoresis (PCR-DGGE) were Thermoanaerobacterium thermosaccharolyticum and Clostridium sp. The lower HY obtained when the cellulose fraction of SCB was used as the substrate might be due to the presence of lignin in the SCB as well as the presence of Lactobacillus parabuchneri and Lactobacillus rhamnosus in the hydrogen fermentation broth.     

Volatile compounds in Bacillus‐fermented soybeans

Thua nao, a traditional, proteolytic, fermented soybean condiment of northern Thailand, was prepared from cooked whole soybeans by natural flora fermentation. The microbial flora during the fermentation was dominated by Bacillus species. The formation of volatile compounds during the fermentation was studied. In addition, the volatile compounds of two samples of commercial dried thua nao and two samples of commercial Japanese natto were analysed. Fermentation led to a large increase in the concentration of total volatile compounds, from 35 µg kg^?1 wet weight in cooked soybeans to 3500 µg kg^?1 wet weight in 72 h fermented material. The major volatile compounds in fermented beans were 3‐hydroxybutanone (acetoin), 2‐methlybutanoic acid, pyrazines, dimethyl disulphide and 2‐pentylfuran. Sun drying of 72 h fermented material resulted in the loss of 65% of total volatiles, including important aroma compounds. The commercial dried thua nao samples had low concentrations of total volatile compounds (380 µg kg^?1 wet weight). It is suggested that improved drying/preservation methods are needed to retain aroma compounds in the traditional products. The natto samples were devoid of aldehydes, aliphatic acids and esters, and sulphur compounds, whereas the thua nao samples contained a diversity of these compounds. Previous investigators have reported these compounds in natto and it is not possible to suggest the existence of systematic differences between the volatile compounds in traditional thua nao prepared with an undefined, mixed microbial flora and those in natto fermented with Bacillus subtilis. © 2001 Society of Chemical Industry     

Performance and population analysis of hydrogen production from sugarcane juice by non-sterile continuous stirred tank reactor augmented with Clostridium butyricum

Non-sterile operation of continuous stirred tank reactor (CSTR) augmented with Clostridium butyricum and fed with sugarcane juice was studied at various hydraulic retention time (HRT). The maximum hydrogen production rate and yield of 3.38 mmol H₂/L/h and 1.0 mol H₂/mol hexose consumed, respectively, were achieved at HRT 4 h. The relationship of the augmented microorganism and normal flora in the fermentation system under non-sterile condition were analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Initially, at HRT 36 h, other species related to Lactobacillus harbinensis and Klebseilla pneumoniae were present as a major group in the reactor. When HRT was decreased to 12, 6 and 4 h, C. butyricum was present with a competition between L. harbinensis and K. pneumoniae. Results indicated that augmented C. butyricum could compete with contaminated microorganisms during non-sterile operation at low HRT (12-4 h) with the support of normal flora (K. pneumoniae).     

Simultaneous production of hydrogen and ethanol from waste glycerol by Enterobacter aerogenes KKU-S1

Factors affecting simultaneous hydrogen and ethanol production from waste glycerol by a newly isolated bacterium Enterobacter aerogenes KKU-S1 were investigated employing response surface methodology (RSM) with central composite design (CCD). The Plackett-Burman design was first used to screen the factors influencing simultaneous hydrogen and ethanol production, i.e., initial pH, temperature, amount of vitamin solution, yeast extract (YE) concentration and glycerol concentration. Results indicated that initial pH, temperature, YE concentration, and glycerol concentration had a statistically significant effect (p ≤ 0.05) on hydrogen production rate (HPR) and ethanol production. The significant factors were further optimized using CCD. Optimum conditions for simultaneously maximizing HPR and ethanol production were YE concentration of 1.00 g/L, glycerol concentration of 37 g/L, initial pH of 8.14, and temperature of 37 °C in which a maximum HPR and ethanol production of 0.24 mmol H₂/L h and 120 mmol/L were achieved.     

Repeated batch fermentation for photo-hydrogen and lipid production from wastewater of a sugar manufacturing plant

Hydrogen and lipid production from sugar manufacturing plant wastewater (SMW) by Rhodobacter sp. KKU-PS1 were investigated. Aji-L (i.e., a waste from the process of crystallizing monosodium glutamate) was used as nitrogen source. Batch fermentation was conducted in 300 mL serum bottles with the working volume of 180 mL to investigate the optimal inoculum size by varying the initial inoculum concentration from 0.23 to 0.92 g_CDW/L. The photo-fermentation was conducted at an initial pH 7.0 and 25.6 °C with continuously light illumination at 7500 lux. The optimal inoculum size of 0.77 g_CDW/L gave the hydrogen production rate (R_m) and lipid production of 5.24 mL H₂/L.h and 407 mg lipid/L, respectively. The hydrogen production from SMW was further examined in 1.7-L photo-bioreactor with the working volume of 1.2-L using the optimal condition from batch experiment. A photo-bioreactor yielded 1.73 times higher R_m than that obtained from the fermentation in serum bottles with a greater lipid production of 424 mg lipid/L. Hydrogen production from SMW in the repeated-batch fermentation was further studied by varying the medium replacement ratios of 25, 50–75%. A maximum biomass and lipid concentration of 2.83 g_CDW/L and 685 mg lipid/L, respectively were achieved at a medium replacement ratio of 75%. C18:1 (51.2%), C18:0 (24.9%) and C16:0 (9.1%) were found as the major free fatty acid. Lactic acid followed by propionic, acetic and butyric acids containing in SMW were the suitable carbon source for biomass production of KKU-PS1.     

Two-stage thermophilic bio-hydrogen and methane production from lime-pretreated oil palm trunk by simultaneous saccharification and fermentation

A simultaneous saccharification and fermentation (SSF) process was applied for thermophilic bio-hydrogen production from lime-pretreated oil palm trunk (OPT) by Thermoanaerobacterium thermosaccharolyticum KKU19. The SSF hydrogen fermentation conditions were optimized to maximize hydrogen yield (HY). Based on Plackett-Burman design, substrate loading and initial pH had significant effects on HY. The substrate loading and initial pH were further optimized using response surface methodology with a central composite design. The optimum conditions were a substrate loading, enzyme loading, inoculum concentration, initial pH and temperature of 4.6%, 10 filter paper unit (FPU)/g-OPT, 10% (v/v), 6.3 and 50 °C, respectively, which yielded the highest HY of 60.22 mL H₂/g-OPT. Structural analysis showed that lime pretreatment and SSF decreased the crystallinity of OPT. Methane production was carried out following the hydrogen production to improve the energy yield from OPT. The results showed that methane production increased total energy yield from 0.65 to 11.79 kJ/g-OPT under the optimal conditions.