Biohydrogen Production in Substrates Combination of Vinasse and Tofu Whey Using Photosynthetic Bacteria Rhodobium marinum

- Nusaibah, Khaswar Syamsu, Dwi Susilaningsih


Hydrogen gas is generated by the fossil fuel refinery process, fixation of sunlight, wind turbine, and water electrolysis. Hydrogen gas is promising energy due to its high energy content and clean combustion. It is also able to be produced by fermentation series, using waste as a substrate. Vinasse is a waste of ethanol distillation from fermented molasses, which still has high Chemical Oxygen Demand (COD) content. The COD is mainly composed of many organic loads that potential to produce hydrogen. Tofu whey (TW), which still has high nitrogen content, is waste generated from the tofu production process. TW is reported to contain high of valuable nutrients, including protein that contains nitrogen. The combination of both liquid wastes is a suitable substrate for hydrogen gas production using Rhodobium marinum. This work aimed to study hydrogen gas, Hydrogen Production Rate, and COD Removal Rate of hydrogen production using a combination of both liquid wastes. Biohydrogen production was examined by varying COD of vinasse/Nitrogen from TW (10,000/1-50,000/1) at a particular fermentation time in the 3rd, 6th, and 9th days. The highest hydrogen gas and Hydrogen Production Rate (HPR) was obtained at substrate concentration 40,000/1 on 9th day of fermentation, namely 95.72±6.51 mL H2/L and 121.44 mL H2/L/d, respectively. COD removal and COD Removal Rate were 7820±400.69 mg COD/L and 799.77 mg COD/L/d. Thus, it can be concluded that the combination of Vinasse and TW has potential as a substrate to biohydrogen production using Rhodobium marinum.


biohydrogen; vinasse; tofu whey; R. marinum.

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I. K. Kapdan and F. Kargi, “Bio-hydrogen production from waste materials,†Enzyme Microb. Technol., vol. 38, no. 5, pp. 569–582, 2006.

M. M. Arimi, J. Knodel, A. Kiprop, S. S. Namango, Y. Zhang, and S. U. Geißen, “Strategies for improvement of biohydrogen production from organic-rich wastewater: A review,†Biomass and Bioenergy, vol. 75, no. 0, pp. 101–118, 2015.

A. Ghimire et al., “A review on dark fermentative biohydrogen production from organic biomass: Process parameters and use of by-products,†Appl. Energy, vol. 144, pp. 73–95, 2015.

D. Das and T. N. Veziroglu, “Advances in biological hydrogen production processes,†Int. J. Hydrogen Energy, vol. 33, no. 21, pp. 6046–6057, 2008.

J. B. Veeramalini, I. A. Ebenezer, S. Park, and J. Jayamuthunagai, “Bioresource Technology Continuous production of biohydrogen from brewery e ffl uent using co- culture of mutated Rhodobacter M 19 and Enterobacter aerogenes,†Bioresour. Technol., vol. 286, no. March, p. 121402, 2019.

O. García-depraect, I. Valdez-vázquez, E. R. Rene, J. Gómez-romero, A. López-lópez, and E. León-becerril, “Lactate- and acetate-based biohydrogen production through dark co-fermentation of tequila vinasse and nixtamalization wastewater: Metabolic and microbial community dynamics,†Bioresour. Technol., 2019.

J. R. Banu, “Industrial Wastewater to Biohydrogen: Possibilities towards successful biorefinery Corresponding Author:†2019.

I. Syaichurrozi, Budiyono, and S. Sumardiono, “Predicting kinetic model of biogas production and biodegradability organic materials: Biogas production from vinasse at variation of COD/N ratio,†Bioresour. Technol., vol. 149, pp. 390–397, 2013.

T. Nandy, S. Shastry, and S. N. Kaul, “Wastewater management in a cane molasses distillery involving bioresource recovery,†2002.

D. Pant and A. Adholeya, “Biological approaches for treatment of distillery wastewater: A review,†vol. 98, pp. 2321–2334, 2007.

G. Vaccari, E. Tamburini, G. Sgualdino, K. Urbaniec, and J. Klemeš, “Overview of the environmental problems in beet sugar processing: Possible solutions,†J. Clean. Prod., vol. 13, no. 5 SPEC. ISS., pp. 499–507, 2005.

J. Chua and S. Liu, “Trends in Food Science & Technology Soy whey: More than just wastewater from tofu and soy protein isolate industry,†Trends Food Sci. Technol., vol. 91, no. June, pp. 24–32, 2019.

J. S. Henao, J. R. Wagner, and G. G. Palazolo, “In fl uence of chemical composition and structural properties on the surface behavior and foam properties of tofu-whey concentrates in acid medium,†vol. 128, no. October 2019, 2020.

M. Kim and D. Lee, “Bioresource Technology Fermentative hydrogen production from tofu-processing waste and anaerobic digester sludge using microbial consortium,†Bioresour. Technol., vol. 101, no. 1, pp. S48–S52, 2010.

C. Lay, B. Sen, S. Huang, C. Chen, and C. Lin, “Sustainable bioenergy production from tofu-processing wastewater by anaerobic hydrogen fermentation for onsite energy recovery,†vol. 58, pp. 60–67, 2013.

K. Anam, M. S. Habibi, T. U. Harwati, and D. Susilaningsih, “Photofermentative hydrogen production using Rhodobium marinum from bagasse and soy sauce wastewater,†Int. J. Hydrogen Energy, vol. 37, no. 20, pp. 15436–15442, 2012.

C. M. Dos Reis, M. F. Carosia, I. K. Sakamoto, M. B. Amâncio Varesche, and E. L. Silva, “Evaluation of hydrogen and methane production from sugarcane vinasse in an anaerobic fluidized bed reactor,†Int. J. Hydrogen Energy, vol. 40, no. 27, pp. 8498–8509, 2015.

R. Ramos-ibarra, R. Garcı, G. Guatemala-morales, E. Arriola-guevara, and G. Toriz-gonz, “ScienceDirect Photofermentation of tequila vinasses by Rhodopseudomonas pseudopalustris to produce hydrogen,†pp. 1–13, 2018.

O. García-depraect and E. León-becerril, “Fermentative biohydrogen production from tequila vinasse via the lactate- acetate pathway: Operational performance, kinetic analysis and microbial ecology,†Fuel, vol. 234, no. June, pp. 151–160, 2018.

M. Tena and R. Solera, “E ff ects of several inocula on the biochemical hydrogen potential of sludge- vinasse co-digestion,†vol. 258, no. July, 2019.

M. Corzo-martinez, G. García-campos, A. Montilla, and F. J. Moreno, “Tofu whey permeate is an efficient source to enzymatically produce prebiotic fructooligosaccharides and novel fructosylated # -galactosides,†2016.

C. Zhao, P. Kim, and J. Eun, “LWT - Food Science and Technology In fl uence of high-intensity ultrasound application on the physicochemical properties, iso fl avone composition, and antioxidant activity of tofu whey,†vol. 117, no. August 2019, 2020.

G. Buitrón and C. Carvajal, “Biohydrogen production from Tequila vinasses in an anaerobic sequencing batch reactor: Effect of initial substrate concentration, temperature and hydraulic retention time,†Bioresour. Technol., vol. 101, no. 23, pp. 9071–9077, 2010.

O. García-depraect, E. R. Rene, and V. F. Diaz-cruces, “Department of Environmental Technology, Centro de Investigación y Asistencia en Department of Environmental Engineering and Water Technology, UNESCO-IHE Corresponding author:†Bioresour. Technol., 2018.

J. Park, D. Kim, S. Kim, J. Yoon, and H. Park, “ScienceDirect Effect of substrate concentration on the competition between Clostridium and Lactobacillus during biohydrogen production,†Int. J. Hydrogen Energy, pp. 1–10, 2017.

C. Z. Lazaro, V. Perna, C. Etchebehere, and M. B. A. Varesche, “Sugarcane vinasse as substrate for fermentative hydrogen production: The effects of temperature and substrate concentration,†Int. J. Hydrogen Energy, vol. 39, no. 12, pp. 6407–6418, 2014.

D. H. Kim, S. H. Kim, I. B. Ko, C. Y. Lee, and H. S. Shin, “Start-up strategy for continuous fermentative hydrogen production: Early switchover from batch to continuous operation,†Int. J. Hydrogen Energy, vol. 33, no. 5, pp. 1532–1541, 2008.

E. Elbeshbishy, B. R. Dhar, G. Nakhla, and H. Lee, “A critical review on inhibition of dark biohydrogen fermentation,†Renew. Sustain. Energy Rev., vol. 79, no. May, pp. 656–668, 2017.

S. Sung, “Ammonia inhibition on thermophilic anaerobic digestion,†vol. 53, pp. 43–52, 2003.

M. Soubes, L. Muxf, A. Fernandez, and S. Tarlera, “Volume 16 No . 2 (February 1994) pp. 195-200 Received 5th January,†vol. 16, no. 2, pp. 195–200, 1994.

X. Chen, Y. Sun, Z. Xiu, X. Li, and D. Zhang, “Stoichiometric analysis of biological hydrogen production by fermentative bacteria,†vol. 31, pp. 539–549, 2006.

A. D. N. Ferraz Júnior, C. Etchebehere, and M. Zaiat, “Mesophilic hydrogen production in acidogenic packed-bed reactors (APBR) using raw sugarcane vinasse as substrate: Influence of support materials,†Anaerobe, vol. 34, pp. 94–105, 2015.

G. Buitrón, G. Kumar, A. Martinez-Arce, and G. Moreno, “Hydrogen and methane production via a two-stage processes (H2-SBR + CH4-UASB) using tequila vinasses,†Int. J. Hydrogen Energy, vol. 39, no. 33, pp. 19249–19255, 2014.

F. Do Carmo Lamaison, P. A. M. De Andrade, A. D. Bigaton, F. D. Andreote, R. V. Antônio, and V. Reginatto, “Long-term effect of acid and heat pretreatment of sludge from a sugarcane vinasse treatment plant on the microbial community and on thermophilic biohydrogen production,†Int. J. Hydrogen Energy, vol. 40, no. 41, pp. 14124–14133, 2015.

D. Kim, D. Lee, and M. Kim, “Enhanced biohydrogen production from tofu residue by acid / base pretreatment and sewage sludge addition,†Int. J. Hydrogen Energy, vol. 36, no. 21, pp. 13922–13927, 2011.

M. Kim, D. Lee, and D. Kim, “Continuous hydrogen production from tofu processing waste using anaerobic mixed microflora under thermophilic conditions,†Int. J. Hydrogen Energy, vol. 36, no. 14, pp. 8712–8718, 2010.

G. Yang and J. Wang, “Co-fermentation of sewage sludge with ryegrass for enhancing hydrogen production: Performance evaluation and kinetic analysis,†Bioresour. Technol., 2017.

L. D. M. Torquato et al., “Potential of biohydrogen production from effluents of citrus processing industry using anaerobic bacteria from sewage sludge,†Waste Manag., 2016.

M. S. Silva, L. B. Oliveira, and C. F. Mahler, “ScienceDirect Hydrogen production through anaerobic co-digestion of food waste and crude glycerol at mesophilic conditions,†pp. 1–10, 2017.

F. M. S. Silva, C. F. Mahler, L. B. Oliveira, and J. P. Bassin, “Hydrogen and methane production in a two-stage anaerobic digestion system by co-digestion of food waste, sewage sludge and glycerol,†2018.



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