• Transactions of the Korean hydrogen and new energy society
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• v.26 no.3
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• pp.247-259
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• 2015

This study was performed to investigate the application of dark $H_2$ fermentation to two-stage bioprocesses for organic waste treatment and energy production. We reviewed information about the two-stage bioprocesses combining dark $H_2$ fermentation with $CH_4$ fermentation, photo $H_2$ fermentation, microbial fuel cells (MFCs), or microbial electrolysis cells (MECs) by using academic information databases and university libraries. Dark fermentative bacteria use organic waste as the sole source of electrons and energy, converting it into $H_2$. The reactions related to dark $H_2$ fermentation are rapid and do not require sunlight, making them useful for treating organic waste. However, the degradation is not complete and organic acids remain. Thus, dark $H_2$ fermentation should be combined with a post-treatment process, such as $CH_4$ fermentation, photo $H_2$ fermentation, MFCs, or MECs. So far, dark $H_2$ fermentation followed by $CH_4$ fermentation is a promising two-stage bioprocess among them. However, if the problems of manufacturing expenses, operational cost, scale-up, and practical applications will be solved, the two-stage bioprocesses combining dark $H_2$ fermentation with photo $H_2$ fermentation, MFCs, or MECs have also infinite potential in organic waste treatment and energy production. This paper demonstrated the feasibility of two-stage bioprocesses combining dark $H_2$ fermentation as a novel system for organic waste treatment and energy production.

• Transactions of the Korean hydrogen and new energy society
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• v.22 no.3
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• pp.333-339
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• 2011

In the present work, it was attempted to produce $H_2$ from food waste by the two-stage fermentation system. Food waste was acidified to lactate by using indigenous lactic acid bacteria under mesophilic condition, and the lactate fermentation effluent (LFE) was subsequently converted to $H_2$ by photo-fermentation. $Rhodobacter$ $sphaeroides$ KD131 was used as the photo-fermenting bacteria. The optimal conditions for lactate fermentation were found to be pH of 5.5 and substrate concentration of 30 g Carbo. COD/L, under which yielded 1.6 mol lactate/mol glucose. By filtering the LFE and adding trace metal, $H_2$ production increased by more than three times compared to using raw LFE, and finally reached the $H_2$ yield of 3.6 mol $H_2$/mol lactate. Via the developed two-stage fermentation system $H_2$ yield of 5.8 mol $H_2$/mol glucose was achieved from food waste, whose value was the highest that ever recorded.

• Transactions of the Korean hydrogen and new energy society
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• v.11 no.1
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• pp.29-41
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• 2000

Anaerobic fermentation using Clostridium butyricum NCIB 9576, and photo-fermentation using Rhodopseudomonas sphaeroides E15-1 were studied for the production of hydrogen from Makkoli, fruits (orange & apple, watermelon & melon) and Tofu wastewaters. From the Makkoli wastewater, which contained $0.94g/{\ell}$ sugars and $2.74g/{\ell}$ soluble starch, approximately $49mM\;H_2/{\ell}$ wastewater was produced during the initial 18h of the anaerobic fermentation with pH control between 6.5-7.0. Several organic acids such as butyric acid, acetic acid, propionic acid, lactic acid and ethanol were also produced. From Watemlelon and melon wastewater, which contained $43g/{\ell}$ sugars, generated about approximately $71mM\;H_2/{\ell}$ wastewater was produced during the initial 24 h of the anaerobic fermentation. Tofu wastewater, pH 6.5, containing $12.6g/{\ell}$ soluble starch and $0.74g/{\ell}$ sugars, generated about $30mM\;H_2/{\ell}$ wastewater, along with some organic acids, during the initial 24 h of anaerobic fermentation. Makkoli and Tofu wastewaters as substrates for the photo-fermentation by Rhodopseudomonas sphaeroides E15-1 produced approximately 37.9 and $22.2{\mu}M\;H_2/m{\ell}$ wastewaters, respectively for 9 days of incubation under the average of 9,000-10,000 lux illumination at the surface of reactor using tungsten halogen lamps. Orange and apple wastewater, which contained 93.4 g/l, produced approximately $13.1{\mu}M\;H_2/m{\ell}$ wastewater only for 2 days of photo-fermentation and the growth of Rhodopseudomonas sphaeroides E15-1 and hydrogen production were stopped.

• Journal of Microbiology and Biotechnology
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• v.22 no.3
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• pp.400-406
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• 2012

To improve the hydrogen yield from biological fermentation of organic wastewater, a co-culture system of dark- and photo-fermentation bacteria was investigated. In a pure-culture system of the dark-fermentation bacterium Clostridium butyricum, a pH of 6.25 was found to be optimal, resulting in a hydrogen production rate of 18.7 ml-$H_2/l/h$. On the other hand, the photosynthetic bacterium Rhodobacter sphaeroides could produce the most hydrogen at 1.81mol-$H_2/mol$-glucose at pH 7.0. The maximum specific growth rate of R. sphaeroides was determined to be 2.93 $h^{-1}$ when acetic acid was used as the carbon source, a result that was significantly higher than that obtained using either glucose or a mixture of volatile fatty acids (VFAs). Acetic acid best supported R. sphaeroides cell growth but not hydrogen production. In the co-culture system with glucose, hydrogen could be steadily produced without any lag phase. There were distinguishable inflection points in a plot of accumulated hydrogen over time, resulting from the dynamic production or consumption of VFAs by the interaction between the dark- and photo-fermentation bacteria. Lastly, the hydrogen production rate of a repeated fed-batch run was 15.9 ml-$H_2/l/h$, which was achievable in a sustainable manner.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.422-427
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• 2003

Rhodopseudomonas palustris P4 can produce $H_2$ either from CO by water-gas shift reaction or from various sugars by anaerobic fermentation. Fermentative $H_2$ production by P4 is fast, but its yield is relatively low due to the formation of various organic acids. In order to increase $H_2$ production yield from glucose, P4 was investigated for the photo-fermentation of acetate which is a major by-product of fermentative $H_2$ production. Experiments were performed in batch modes using both light-grown and dark-grown cells. When the dark-grown P4 was challenged with light and acetate, $H_2$ was produced with the consumption of acetate after a lag period of 25 h. $H_2$ production was inhibited when a nitrogen source, especially ammonium, is present. When the dark-fermentation broth containing acetate was adopted for photo-fermentation with light-grown cells, $H_2$ production and concomitant acetate consumption occurred without a lag period. The $H_2$ yield was estimated as 2.4 - 2.8 mol $H_2/mol$ acetate and the specific $H_2$ production rate was as 9.8 ml $H_2/g$ cell${\cdot}$h, The fact that a single strain can perform both dark- and light-fermentation gives a great advantage in process development Compared to a one-step dark-fermentation, the combined dark- and light-fermentation can increase the $H_2$ production yield on glucose by two-fold.

• Journal of Energy Engineering
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• v.15 no.2 s.46
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• pp.118-126
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• 2006

This publication provides an overview of the state-of-the-art and perspective of biological $H_2$ production from water and/or organic substances. The biological $H_2$ production processes, being explored in fundamental and applied researches, are direct and indirect biophotolysis from water, photo-fermentation, dark anaerobic fermentation and in vitro $H_2$ production. The development of biological $H_2$ production technology, as an energy carrier, started at the late 1940's in the lab-scale. Now it has a high priority in the world, especially USA, Japan, EU and Korea.

• KSBB Journal
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• v.20 no.6
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• pp.393-400
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• 2005

Hydrogen($H_2$) as a clean, and renewable energy carrier will be served an important role in the future energy economy. Several biological $H_2$ production processes are known and currently under development, ranging from direct bio-photolysis of water by green algae, indirect bio-photolysis by cyanobacteria including the separated two stage photolysis using the combination of green algae and photosynthetic microorganisms or green algae alone, dark anaerobic fermentation by fermentative bacteria, photo-fermentation by purple bacteria, and water gas shift reaction by photosynthetic or fermentative bacteria. In this paper, biological $H_2$ production processes, that are being explored in fundamental and applied research, are reviewed.

• Journal of Korean Society on Water Environment
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• v.27 no.6
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• pp.926-931
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• 2011

Among various techniques for hydrogen production from organic wastewater, a dark fermentation is considered to be the most feasible process due to the rapid hydrogen production rate. However, the main drawback of it is the low hydrogen production yield due to intermediate products such as organic acids. To improve the hydrogen production yield, a co-culture system of dark and photo fermentation bacteria was applied to this research. The maximum specific growth rate of R. sphaeroides was determined to be $2.93h^{-1}$ when acetic acid was used as a carbon source. It was quite high compared to that of using a mixture of volatile fatty acids (VFAs). Acetic acid was the most attractive to the cell growth of R. sphaeroides, however, not less efficient in the hydrogen production. In the co-culture system with glucose, hydrogen could be steadily produced without any lag-phase. There were distinguishable inflection points in the accumulation of hydrogen production graph that resulted from the dynamic production of VFAs or consumption of it by the interaction between the dark and photo fermentation bacteria. Lastly, the hydrogen production rate of a repeated fed-batch run was $15.9mL-H_2/L/h$, which was achievable in the sustainable hydrogen production.

• Transactions of the Korean hydrogen and new energy society
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• v.22 no.4
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• pp.451-457
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• 2011

Purple non-sulfur (PNS) bacterium $Rhodobacter$ $sphaeroides$ KD131 was studied with the aim of achieving maximum hydrogen production using various carbon and nitrogen sources at different pH conditions. Cells grew well and produced hydrogen using $(NH_4){_2}SO_4$ or glutamate as a nitrogen source in combination with a carbon substrate, succinate or malate. During 48h of photo-heterotrophic fermentation under 110$W/m^2$ illumination using a halogen lamp at $30^{\circ}C$, 67% of 30mM succinate added was degraded and the hydrogen yield was estimated as 3.29mol $H^2$/mol-succinate. However, less than 30% of formate was consumed and hydrogen was not produced due to a lack of genes coding for the formate-hydrogen lyase complex of strain KD131. Initial cell concentrations of more than 0.6g dry cell weight/L-culture broth were not favorable for hydrogen evolution by cell aggregation, thus leading to substrate and light unavailability. In a modified Sistrom's medium containing 30mM succinate with a carbon to nitrogen ratio of 12.85 (w/w), glutamate produced 1.40-fold more hydrogen compared to ammonium sulfate during the first 48h. However, ammonium sulfate was 1.78-fold more effective for extended cultivation of 96h. An initial pH range from 6.0 to 9.0 influenced cell growth and hydrogen production, and maintenance of pH 7.5 during photofermentation led to the increased hydrogen yield.

• Transactions of the Korean hydrogen and new energy society
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• v.21 no.2
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• pp.136-142
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• 2010

Photobiological $H_2$ production was compared using purple non-sulfur bacteria Rhodobacter sphaeroides KD131 in the medium containing various organic acids as the carbon source and electron doner under illumination of $110\;W/m^2$ using halogen lamp at $30^{\circ}C$. The organic acids used were 0~120 mM acetate, butyrate, lactate and malate. Initial pH 7.0 and cell concentration 1.0 at 660nm were increased to pH 8 and 4.4~5.1, respectively during 24hrs of photo-fermentation when lactate and malate were used. However, acetate and butyrate increased pH to 9 and cell concentration to 3.2~3.9 of malate at the same experimental conditions. Optimum ranges of organic acids concentration and carbon/nitrogen ratio were 30~60 mM and 10~20, respectively. When malate was used as the substrate, maximum $H_2$ production 1.1 ml $H_2$/ml broth, which is equivalent to 1.97 mol $H_2$/mol malate was observed.