Biohythane production from palm oil mill effluent – a preliminary evaluation of a two-stage anaerobic digestion

@#Aims:This research aims to investigate the potential of biohythane (biohydrogen and biomethane) production from palm oil mill effluent (POME) in a two-stage anaerobic digestion (AD) system.Methodology and results:A two-stage AD system was configured with a thermophilic dark fermenter (TDF) for biohydrogen (H2) production and a thermophilic anaerobic contact digester (TACD) for biomethane (CH4) production. To adjust pH 5.5 for dark fermentation, the settled sludge was recirculated from TACD to TDF. The hydraulic retention time (HRT) applied in TDF and TACD was 3.75 and 6.25 day, respectively.Conclusion, significance and impact of study:The sludge recirculation from TACD was able to adjust the pH in TDF to the optimum value of 5.5. The total COD and TSSdegradation were 63.12 and 77.94 %, respectively. The H2production in TDF was 1.54 L H2/L POME and the CH4production in TACD was 19.87 L CH4/L POME. The H2and CH4yielded 0.085 L/g CODremovedand 0.339 L/g CODremoved, respectively, with total energy recovery equivalent to 661.02 MJ/m3POME. Only 2.28 % of this energy was contributed by H2and the remaining was dominated by CH4.

Two phases fermentative process for hydrogen and methane production from cassava wastewater / Processo fermentativo em duas fases para a produção de hidrogênio e metano a partir da manipueira

Introduction: Hydrogen and methane production was investigated in two phases of fermentative process. Objective: At the acidogenic phase, an anaerobic fluidized bed reactor was fed with cassava wastewater producing hydrogen. Methods: Expanded clay was used as a support material for biomass immobilization. The reactor was operated with HRT ranging from 8-1 h. Results: The best hydrogen yield production was 1.91 mol H2/mol glucose at HRT of 2 h. At the methanogenic phase, the acidogenic process effluent fed a fixed-bed reactor producing methane. Conclusion: Sururu (Mytella falcata) shells was used as support acted as pH neutralizer in the fixed-bed reactor, yielding best (0.430±0.150 Lmethane/gCOD) with 12h HRT phase. (AU)

Introdução: A produção de hidrogênio e metano foi avaliada em um processo fermentativo de duas fases. Objetivo: Na fase acidogênica, um reator anaeróbio de leito fluidificado foi alimentado com manipueira para a produção de hidrogênio. Métodos: Argila expandida foi utilizada com material suporte para a adesão microbiana. O reator foi operado com TDH, variando entre 8-1h. Resultados: O melhor rendimento de produção de hidrogênio foi 1.91 mol, H2/mol glicose em TDH de 2 h. Na fase metanogênica, o efluente do processo acidogênico alimentou um reator de leito fixo para a produção de metano. Conclusão: Conchas de Sururo (Mytella falcata) foram utilizadas como suporte, atuando como neutralizador do pH no reator de leito fixo, melhor rendimento (0.430±0.150 Lmethane/gDQO) na fase com TDH de 12h. (AU)

Enhancement of biohydrogen production from starch processing wastewater and further inside its ecosystem disclosed by 16S rDNA sequencing and FISH

ABSTRACT Biohydrogen production from starch processing wastewater in this study resulted the highest yield of 61.75 mL H2/g COD at initial pH 7.0, thermophilic temperature, and iron concentration 800 mg Fe/L. The yield was 2-folded higher than the operation at mesophilic temperature or without iron addition. Cell immobilization by addition of biomaterials (BM) could improve the hydrogen yield by 2-folded comparing to the non-addition. BM from plants (loofa sponge) was found producing higher yield than that from animals (silk cocoon), and optimal concentration of BM was 5% (V/V). Furthermore, it was revealed further inside its ecosystem using SEM, 16S rDNA sequencing and FISH. There was found rod-shaped microorganisms of Bacillus cereus, which reported as efficient starch-utilizing hydrogen producers, was dominant in the system with population of 47% of all specie identified.

Buffering action of acetate on hydrogen production by Ethanoligenens harbinense B49

The buffering effect of acetate on hydrogen production during glucose fermentation by Ethanoligenens harbinense B49 was investigated compared to phosphate, a widely used fermentative hydrogen production buffer. Specific concentrations of sodium acetate or phosphate were added to batch cultures, and the effects on hydrogen production were comparatively analyzed using a modified Gompertz model. Adding 50 mM acetate or phosphate suppressed the hydrogen production peak and slightly extended the lag phase. However, the overall hydrogen yields were 113.5 and 108.5 mmol/L, respectively, and the final pH was effectively controlled. Acetate buffered against hydrogen production more effectively than did phosphate, promoting cell growth and preventing decreased pH. At buffer concentrations 100-250 mM, the maximum hydrogen production was barely suppressed, and the lag phase extended past 7 h. Therefore, although acetate inhibits hydrogen production, using acetate as a buffer (like phosphate) effectively prevented pH drops and increased substrate consumption, enhancing hydrogen production.

Immobilized Biofilm in Thermophilic Biohydrogen Production using Synthetic versus Biological Materials

Biohydrogen production was studied from the vermicelli processing wastewater using synthetic and biological materials as immobilizing substrate employing a mixed culture in a batch reactor operated at the initial pH 6.0 and thermophilic condition (55 ± 1ºC). Maximum cumulative hydrogen production (1,210 mL H2/L wastewater) was observed at 5% (v/v) addition of ring-shaped synthetic material, which was the ring-shaped hydrophobic acrylic. Regarding 5% (v/v) addition of synthetic and biological materials, the maximum cumulative hydrogen production using immobilizing synthetic material of ball-shaped hydrophobic polyethylene (HBPE) (1,256.5 mL H2/L wastewater) was a two-fold increase of cumulative hydrogen production when compared to its production using immobilizing biological material of rope-shaped hydrophilic ramie (609.8 mL H2/L wastewater). SEM observation of immobilized biofilm on a ball-shaped HBPE or a rope-shaped hydrophilic ramie was the rod shape and gathered into group.

Feasibility of biohydrogen production from industrial wastes using defined microbial co-culture

BACKGROUND: The development of clean or novel alternative energy has become a global trend that will shape the future of energy. In the present study, 3 microbial strains with different oxygen requirements, including Clostridium acetobutylicum ATCC 824, Enterobacter cloacae ATCC 13047 and Kluyveromyces marxianus 15D, were used to construct a hydrogen production system that was composed of a mixed aerobic-facultative anaerobic-anaerobic consortium. The effects of metal ions, organic acids and carbohydrate substrates on this system were analyzed and compared using electrochemical and kinetic assays. It was then tested using small-scale experiments to evaluate its ability to convert starch in 5 L of organic wastewater into hydrogen. For the one-step biohydrogen production experiment, H1 medium (nutrient broth and potato dextrose broth) was mixed directly with GAM broth to generate H2 medium (H1 medium and GAM broth). Finally, Clostridium acetobutylicum ATCC 824, Enterobacter cloacae ATCC 13047 and Kluyveromyces marxianus 15D of three species microbial co-culture to produce hydrogen under anaerobic conditions. For the two-step biohydrogen production experiment, the H1 medium, after cultured the microbial strains Enterobacter cloacae ATCC 13047 and Kluyveromyces marxianus 15D, was centrifuged to remove the microbial cells and then mixed with GAM broth (H2 medium). Afterward, the bacterial strain Clostridium acetobutylicum ATCC 824 was inoculated into the H2 medium to produce hydrogen by anaerobic fermentation. RESULTS: The experimental results demonstrated that the optimum conditions for the small-scale fermentative hydrogen production system were at pH 7.0, 35°C, a mixed medium, including H1 medium and H2 medium with 0.50 mol/L ferrous chloride, 0.50 mol/L magnesium sulfate, 0.50 mol/L potassium chloride, 1% w/v citric acid, 5% w/v fructose and 5% w/v glucose. The overall hydrogen production efficiency in the shake flask fermentation group was 33.7 mL/h-1.L-1, and those the two-step and the one-step processes of the small-scale fermentative hydrogen production system were 41.2 mLVh-1.L-1 and 35.1 mL/h-1.L-1, respectively. CONCLUSION: Therefore, the results indicate that the hydrogen production efficiency of the two-step process is higher than that of the one-step process.

Profiling carbohydrate composition, biohydrogen capacity, and disease resistance in potato

Background: Potato (Solanum tuberosum) is one of the most important sources of carbohydrates in human diet. Because of its high carbohydrate levels it recently has also received attention in biohydrogen production. To exploit the natural variation of potato with respect to resistance to major diseases, carbohydrate levels and composition, and capacity for biohydrogen production we analyzed tubers of native, improved, and genetically modified potatoes, and two other tuberous species for their glucose, fructose, sucrose, and starch content. Results: High-starch potato varieties were evaluated for their potential for Caldicellulosiruptor saccharolyticus-mediated biohydrogen production with Desirée and Rosita varieties delivering the highest biohydrogen amounts. Native line Vega1 and improved line Yagana were both immune to two isolates (A291, A287) of Phytophthora infestans. Conclusions: Our data demonstrate that native potato varieties might have great potential for further improving the multifaceted use of potato in worldwide food and biohydrogen production.

Biohydrogen production by Thermoanaerobacterium thermosaccharolyticum KKU-ED1: Culture conditions optimization using mixed xylose/arabinose as substrate

Background: Biological hydrogen production by microorganisms can be divided into two main categories i.e. photosynthetic organisms that produce hydrogen using light as energy source and anaerobic bacteria that produce hydrogen via dark fermentation. Dark fermentative hydrogen production by anaerobic bacteria has the advantages of a higher HPR without illumination and of the capability to convert various kinds of substrate. Results: Thermophilic hydrogen producer was isolated from elephant dung and identified as Thermoanaerobacterium thermosaccharolyticum KKU-ED1 by 16S rRNA gene analysis, which was further used to produce hydrogen from mixed pentose sugar i.e., xylose/arabinose. The optimum conditions for hydrogen production from mixed xylose/arabinose by KKU-ED1 were a 1:1 xylose/arabinose mixture at the total concentration of 5 g/L, initial pH of 6.5 and temperature of 55ºC. Under the optimum conditions, hydrogen from sugar derived from acid-hydrolyzed sugarcane bagasse at a reducing sugar concentration were achieved. Soluble metabolite product (SMP) was predominantly acetic acid indicating the acetate-type fermentation. Conclusions: The strain KKU-ED1 appeared to be a suitable candidate for thermophilic fermentative hydrogen production from hemicellulosic fraction of lignocellulosic materials due to its ability to use various types of carbon sources.

Effect of temperature and initial pH on biohydrogen production from palm oil mill effluent: long-term evaluation and microbial community analysis

Anaerobic sludge from palm oil mill effluent (POME) treatment plant was used as a source of inocula for the conversion of POME into hydrogen. Optimization of temperature and initial pH for biohydrogen production from POME was investigated by response surface methodology. Temperature of 60ºC and initial pHof 5.5 was optimized for anaerobic microflora which gave a maximum hydrogen production of 4820 ml H2/l-POME corresponding to hydrogen yield of 243 ml H2/g-sugar. Total sugar consumption and chemical oxygen demand (COD) removal efficiency were 98.7 percent and 46 percent respectively. Long-term hydrogen production in continuous reactor at HRT of 2 days, 1 day and 12 hrs were 4850 +/- 90, 4660 +/- 99 and 2590 +/- 120 ml H2/l-POME, respectively. Phylogenetic analysis of the mixed culture revealed that members involved hydrogen producers in both batch and continuous reactors were phylogenetically related to the Thermoanaerobacterium thermosaccharolyticum. Batch reactor showed more diversity of microorganisms than continuous reactor. Microbial community structure of batch reactor was comprised of T. thermosaccharolyticum, T. bryantii, Thermoanaerobacterium sp., Clostridium thermopalmarium and Clostridium NS5-4, while continuous reactor was comprised of T. thermosaccharolyticum, T. bryantii and Thermoanaerobacterium sp. POME is good substrate for biohydrogen production under thermophilic condition with Thermoanaerobacterium species play an important role in hydrogen fermentation.

Isolation and Characterization of a New High Efficient H2-producing Bacterium Enterococcus sp. LG1 / 微生物学通报

12 strains of H2-producing bacteria were isolated and purified from anaerobic sludge, aerobic sludge and river bottom sludge by Hungate method. A new species of high efficient hydrogen production bacterium Enterococcus sp. LG1 (registration number: EU258743 ) was studied deeply. It was showed that the Enterococcus sp. LG1 was an anaerobic and Gram-negative bacterium. Sequence analysis of this type of clones showed that it was affiliated with the genus Enterococcus and it was not reported yet in other paper at present. Meanwhile, batch tests of anaerobic fermentative hydrogen production by Enterococcus sp. LG1 were investigated by using sterilization pretreated sludge as substrate. The changes of soluble COD, protein, carbohydrate and pH value during hydrogen fermentation were monitored. It was found that only hydrogen and carbon dioxide were produced by this strain and no methane was detected during fermentation. The maximal hydrogen yield was 36.48 mL/g TCOD and the hydrogen concentration in the gas phase was 73.5%. The Enterococcus sp. LG1 was a butyrate fermentation bacteria analyzed by metabolites.

Cloning and Transformation of lba Gene of Glycine max into Chloroplast of Chlamydomonas reinhardtii / 中国生物工程杂志

To decrease the oxygen content in the cell is a key method to improve hydrogen production in Chlamydomonas reinhardtii.A new approach was developed by transforming the leghemoglobin gene lba,which has high affinity to oxygen,into the chloroplast of C.reinhardtii to get a low dissolved oxygen in the cell and result into improvement of H2 ase activity and H2 yield. The results showed that lba was successfully transformed into the chloroplast of C.reinahrdtii strain 849 and did not affect its growth significantly. The work paved the road for further regulation of lba expression in the chloroplast to improve of hydrogen production of C.reinahrdtii.

Combined Mutation Breeding of H_2-producing Strain and Hydrogen Producing Characterization of a H_2-producing Mutant HCM-23 / 微生物学通报

The fermentative H2-producing strain Clostridium sp. H-61 was isolated from anaerobic sludge,was used as an original strain which was induced by NTG and UV for increasing and the hydrogen production ability. One of the highest efficient H2-producing mutants was named as HCM-23 with its stable hydrogen production ability. which was measured in the batch culture experiments. With the condition of 10 g/L glucose,its cumulative hydrogen yield and hydrogen production rate was 3024 mL/L and 33.19 mmol H2/g DW?h,69.89% and 68.14% higher than that of the original strain,respectively. The terminal liquid product compositions showed that the mutant HCM-23 fermentation was ethanol type,while the original strain H-61 fermentation was butyric acid type. Varieties of parameters of hydrogen production fermentation studied,including time,carbon source,nitrogen source,glucose concentration,glucose utilization,initial pH and incubation temperature had been studied,indicated the optimum condition of hydrogen production for the mutantHCM-23 as initial pH 6.5,temperature 36 ℃,and the favorite substrate was sucrose. The hydrogen production characters of the mutant and the original strain were different,such as,the growth lag phase and the utilization of inorganic nitrogen source,etc. This work shows a good application potential of NTG-UV combined mutation in the biohydrogen production. And the hydrogen production mechanism and metabolic pathway should be explored furthermore.