Abstract
Cheese whey-based biohydrogen production was seen in batch experiments via dark fermentation by free and immobilized Enterobacter aerogenes MTCC 2822 followed by photofermentation of VFAs (mainly acetic and butyric acid) in the spent medium by Rhodopseudomonas BHU 01 strain. E. aerogenes free cells grown on cheese whey diluted to 10 g lactose/L, had maximum lactose consumption (∼79%), high production of acetic acid (1,900 mg/L), butyric acid (537.2 mg/L) and H2 yield (2.04 mol/mol lactose; rate,1.09 mmol/L/h). The immobilized cells improved lactose consumption (84%), production of acetic acid (2,100 mg/L), butyric acid (718 mg/L) and also H2 yield (3.50 mol/mol lactose; rate, 1.91 mmol/L/h). E. aerogenes spent medium (10 g lactose/L) when subjected to photofermentation by free Rhodopseudomonas BHU 01 cells, the H2 yield reached 1.63 mol/mol acetic acid (rate, 0.49 mmol/L/h). By contrast, immobilized Rhodopseudomonas cells improved H2 yield to 2.69 mol/mol acetic acid (rate, 1.87 mmol/L/h). The cumulative H2 yield for free and immobilized bacterial cells was 3.40 and 5.88 mol/mol lactose, respectively. Bacterial cells entrapped in alginate, had a sluggish start of H2 production but outperformed the free cells subsequently. Also, the concomitant COD reduction for free cells (29.5%) could be raised to 36.08% by immobilized cells. The data suggest that two-step fermentative H2 production from cheese whey involving immobilized bacterial cells, offers greater substrate to- hydrogen conversion efficiency, and the effective removal of organic load from the wastewater in the long-term.
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Zhu, H., Suzuki, T., Tsygankov, A. A., Asada, Y., & Miyake, J. (2011). Hydrogen production from tofu wastewater by Rhodobacter sphaeroides immobilized in agar gels. International Journal of Hydrogen Energy, 24, 305–310.
Abboud, M. M., Aljundi, I. H., Khleifat, K. M., & Dmour, S. (2010). Biodegradation kinetics and modeling of whey lactose by bacterial hemoglobin VHb-expressing Escherichia coli strain. Biochemical Engineering Journal, 48, 166–172.
Mohan, S. V., Babu, V. L., & Sarma, P. N. (2007). Anaerobic biohydrogen production from dairy waste water treatment in sequencing batch reactor (AnSBR): effect of organic loading rate. Enzyme and Microbial Technology, 41, 506–515.
Ferchichi, M., Crabbe, E., Gil, G.-H., Hintz, W., & Almadidy, A. (2005). Influence of initial pH on hydrogen production from cheese whey. Journal of Biotechnology, 120, 402–409.
Davila-Vazquez, G., Alatriste-Mondragon, F., De Leon-Rodriguez, A., & Razo-Flores, E. (2008). Fermentative hydrogen production in batch experiments using lactose, cheese whey and glucose: Influence of initial substrate concentration and pH. International Journal of Hydrogen Energy, 33, 4989–4997.
Singh, S. P., Srivastava, S. C., & Pandey, K. D. (1994). Hydrogen production by Rhodopseudomonas at the expense of vegetable starch, sugarcane juice and whey. International Journal of Hydrogen Energy, 19, 437–440.
Antonopoulou, G., Stamatelatou, K., Venetsaneas, N., Kornaros, M., & Lyberatos, G. (2008). Biohydrogen and methane production from cheese whey in a two-stage anaerobic process. Industrial and Engineering Chemistry Research, 47, 5227–5233.
Stamatelatou, K., Antonopoulou, G., Tremouli, A., & Lyberatos, G. (2011). Production of gaseous biofuels and electricity from cheese whey. Industrial and Engineering Chemical Research, 50, 639–644.
Azbar, N., Dokgoz, F. T., Keskin, T., Eltem, R., Korkmaz, K. S., Gezgin, Y., Akbal, Z., Oncel, S., Dalay, M. C., & Gonen, C. (2009). Comparative evaluation of bio-hydrogen production from cheese whey waste water under thermophilic and mesophilic anaerobic conditions. International Journal of Green Energy, 6, 192–200.
Song, W., Cheng, J., Zhou, J., Xie, B., Su, H., & Cen, K. (2010). Cogeneration of hydrogen and methane from protein-mixed food waste water by two-phase anaerobic process. International Journal of Hydrogen Energy, 35, 3141–3146.
Alalayah, W. M., Kalil, M. S., Kadhum, A. A. H., Jahim, J. M., Jaapar, S. Z. S., & Alauj, N. M. (2009). Bio-hydrogen production using a two-stage fermentation process. Pakistan Journal of Biological Sciences, 12, 1462–1467.
Ozmihci, S., & Kargi, F. (2010). Bio-hydrogen production by photo-fermentation of dark fermentation effluent with intermittent feeding and effluent removal. International Journal of Hydrogen Energy, 35, 6674–6680.
Singh, S. P., Srivastava, S. C., & Pandey, K. D. (1990). Photoproduction of hydrogen by a non-sulphur bacterium isolated from root zones of water fern Azolla pinnata. International Journal of Hydrogen Energy, 15, 403–406.
Tanisho, S., & Ishiwata, Y. (1995). Continuous hydrogen production from molasses by fermentation using urethane as a support of flocks. International Journal of Hydrogen Energy, 20, 541–545.
Yokoi, H., Tokushige, T., Hirose, J., Hayashi, S., & Takasaki, Y. (1997). Hydrogen production by immobilized cells of aciduric Enterobacter aerogenes strain HO-39. Journal of Fermentation and Bioengineering, 8, 481–484.
Rachmann, M. A., Nkashimada, Y., Kakizono, T., & Nishio, N. (1998). Hydrogen production with high yield and high evolution rate by self-flocculated cells of Enterobacter aerogenes in a packed-bed reactor. Applied Microbiology and Biotechnology, 49, 450–454.
Kumar, N., & Das, D. (2001). Continuous hydrogen production by immobilized Enterobacter cloacae IIT-BT 08 using lignocellulosic materials as solid matrices. Enzyme and Microbial Technology, 29, 280–287.
Liu, X., Zhu, Y., & Yang, S.-T. (2006). Butyric acid and hydrogen production by Clostridium tyrobutyricum ATCC 25755 and mutants. Enzyme and Microbial Technology, 38, 521–528.
Jo, J.-H., Lee, D.-S., Park, D., & Park, J.-M. (2008). Biological hydrogen production by immobilized cells of Clostridium tyrobutyricum JM1 isolated from a food waste treatment process. Bioresource Technology, 99, 6666–6672.
Wang, Y.-Z., Liao, Q., Zhu, X., Tian, X., & Zhang, C. (2010). Characterstics of hydrogen production and substrate consumption of Rhodopseudomonas palustris CQK 01 in an immobilized-cell photobioreactor. Bioresource Technology, 101, 4034–4041.
Liu, B.-F., Xie, G.-J., Guo, W.-Q., Ding, J., & Ren, N.-Q. (2011). Optimization of photo-hydrogen production by immobilized Rhodopseudomonas faecalis RLD-53. Natural Resources, 2, 1–7.
Tian, X., Liao, Q., Liu, W., Wang, Y.-Z., Zhu, X., Li, J., & Wang, H. (2009). Photohydrogen production rate of a PVA-boric acid gel granule containing immobilized photosynthetic bacteria cells. International Journal of Hydrogen Energy, 34, 4708–4717.
Pfennig, N. (1967). Photosynthetic bacteria. Annual Review of Microbiology, 21, 285–324.
Lowry, O. H., Rosebrough, N. J., Farr, A. L., & Randall, R. J. (1951). Protein measurement with the Folin–Phenol reagent. Journal of Biological Chemistry, 193, 265–275.
Herbert, D., Phipps, P. J., & Strange, R. E. (1971). In J. R. Norris & D. W. Ribbons (Eds.), Methods in microbiology, vol.5B: Chemical analysis of microbial cells (pp. 209–344). London: Academic.
Dubois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. A., & Smith, F. (1956). Colorimetric method for determination of sugars and related substances. Analytical Chemistry, 28, 350–356.
APHA. (1995). Standard methods for the examination of water and waste water (19th ed.). USA: American Public Health Association.
Yokoi, H., Tokushige, T., Hirose, J., Hayashi, S., & Takasaki, Y. (1998). H2 production from starch by a mixed culture of Clostridium butyricum and Enterobacter aerogenes. Biotechnology Letters, 20, 143–147.
Papageorgiou, G. C., Kalosaka, K., Sotiropoulou, G., Barbotin, J. N., Thomasett, B., & Thomas, T. (1988). Entrapment of active ion-permeable cyanobacteria (Anacystis nidulans) in calcium alginate. Applied Microbiology and Biotechnology, 29, 565–571.
Fabiano, B., & Perego, P. (2002). Thermodynamic study and optimization of hydrogen production by Enterobacter aerogenes. International Journal of Hydrogen Energy, 27, 149–156.
Oh, Y. K., Seol, E. H., Kim, J. R., & Park, S. (2003). Fermentative biohydrogen production by a new chemoheterotrophic bacterium Citrobacter sp. Y19. International Journal of Hydrogen Energy, 28, 1353–1359.
Ferchichi, M., Crabbe, E., Hintz, W., Gill, G.-H., & Almadidy, A. (2005). Influence of culture parameters on biological hydrogen production by Clostridium saccharoperbutylacetonium ATCC 27021. World Journal of Microbiology and Biotechnology, 21, 855–862.
Calli, B., Schoenmaekers, K., Vanbroekhoven, K., & Diels, L. (2008). Dark fermentative H2 production from xylose and lactose—effects of on-line pH control. International Journal of Hydrogen Energy, 33, 522–530.
Fang, H. H. P., Zhu, H., & Zhang, T. (2006). Phototrophic hydrogen production from glucose by pure and co-cultures of Clostridium butyricum and Rhodobacter sphaeroides. International Journal of Hydrogen Energy, 31, 2223–2230.
Liu, B.-F., Ren, N.-Q., Tang, J., Ding, J., Liu, W.-Z., Xu, J.-F., Cao, G.-L., Guo, W.-Q., & Xie, G.-J. (2010). Biohydrogen production by mixed culture of photo- and dark fermentation bacteria. International Journal of Hydrogen Energy, 35, 2858–2862.
Oh, Y.-K., Seol, E.-H., Kim, M.-S., & Park, S. (2004). Photoproduction of hydrogen from acetate by a chemoheterotrophic bacterium Rhodopseudomonas palustris P4. International Journal of Hydrogen Energy, 29, 1115–1121.
Barbosa, M. J., Rocha, J. M. S., Tramper, J., & Wijffels, R. H. (2001). Acetate as a carbon source for hydrogen production by photosynthetic bacteria. Journal of Biotchnology, 85, 25–33.
Chen, C. Y., Lee, C. M., & Chang, J. S. (2006). Feasibility study on bioreactor strategies for enhanced photohydrogen from Rhodopseudomonas palustris WP 3-5 using optical-fibre assisted illumination systems. International Journal of Hydrogen Energy, 31, 2345–2355.
Ren, N.-Q., Liu, B.-F., Zheng, G.-X., Xing, D.-F., Zhao, X., Guo, W.-Q., & Ding, J. (2009). Strategy for enhancing photo-hydrogen production yield by repeated fed-batch cultures. International Journal of Hydrogen Energy, 34, 7579–7584.
Seifert, K., Waligorska, M., & Laniecki, M. (2010). Hydrogen generation in photobiological process from dairy waste water. International Journal of Hydrogen Energy, 35, 9624–9629.
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This work was supported by grant 103/131/2008-NT from Ministry of New and Renewable Energy, Govt. of India and Hydrogen Energy Centre of this University.
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Rai, P.K., Singh, S.P. & Asthana, R.K. Biohydrogen Production from Cheese Whey Wastewater in a Two-Step Anaerobic Process. Appl Biochem Biotechnol 167, 1540–1549 (2012). https://doi.org/10.1007/s12010-011-9488-4
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DOI: https://doi.org/10.1007/s12010-011-9488-4