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international journal of hydrogen energy 34 (2009) 764–771

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Biohydrogen production by Clostridium butyricum EB6 from palm oil mill effluent
Mei-Ling Chonga, Raha Abdul Rahima, Yoshihito Shiraib, Mohd Ali Hassana,*
Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences,Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia b Graduate School of Life Sciences and System Engineering, Kyushu Institute of Technology, 808-0196 Hibikimo 2-4, Wakamatsu-ku, Kitakyushu-shi, Fukuoka, Japan

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Article history: Received 4 October 2007 Received in revised form 23 June 2008 Accepted 23 October 2008 Available online 11 December 2008 Keywords: BiohydrogenClostridium butyricum EB6 Palm oil mill effluent (POME)

A hydrogen producer was successfully isolated from anaerobic digested palm oil mill effluent (POME) sludge. The strain, designated as Clostridium butyricum EB6, efficiently produced hydrogen concurrently with cell growth. A controlled study was done on a synthetic medium at an initial pH value of 6.0 with 10 g/L glucose with the maximumhydrogen production at 948 mL H2/L-medium and the volumetric hydrogen production rate at 172 mL H2/L-medium/h. The supplementation of yeast extract was shown to have a significant effect with a maximum hydrogen production of 992 mL H2/L-medium at 4 g/L of yeast extract added. The effect of pH on hydrogen production from POME was investigated. Experimental results showed that the optimum hydrogenproduction ability occurred at pH 5.5. The maximum hydrogen production and maximum volumetric hydrogen production rate were at 3195 mL H2/L-medium and 1034 mL H2/L-medium/h, respectively. The hydrogen content in the biogas produced was in the range of 60–70%. ª 2008 International Association for Hydrogen Energy. Published by Elsevier Ltd. All rights reserved.



Biohydrogen hasbeen well documented in recent years and has generated increased attention from researchers due to its characteristics as an ideal, clean and sustainable energy resource for the future. In nature, biohydrogen is produced during acidogenic waste treatment process where acid forming bacteria produces organic acid compound, hydrogen and carbon dioxide [1]. However, no hydrogen is bubbling out due tothe coexisting bacteria that readily consume hydrogen as a source of reducing power. Therefore, a specific environment needs to be created to support the growth of hydrogen producer and reduce the number of hydrogen consumer [2]. For this aim, pretreatment on sludge sample by heat and

chemical method could be carried out to eliminate the presence of methanogens as most of the hydrogen producercould produce endospore that survive at harsh environment [3,4]. Recently, experiments have been carried out to study the possibility of hydrogen production using organic wastes from various industries in combination with the wastewater treatment strategy [5,6]. For example, in Malaysia, the palm oil industry annually generates about 15.2 million tons of wastewater, known as palm oil mill effluent(POME). Due to the nature of POME, with high cellulose and lignocellulosic material, it takes a long time to degrade the organic substances. Previously, research has been carried out to use POME sludge as an inoculum, and it has produced a promising level of hydrogen production [7]. However, the cultivation of

* Corresponding author. Tel.: þ60 3 89467590; fax: þ60 3 89467593. E-mail (M.-L. Chong). 0360-3199/$ – see front matter ª 2008 International Association for Hydrogen Energy. Published by Elsevier Ltd. All rights reserved. doi:10.1016/j.ijhydene.2008.10.095

international journal of hydrogen energy 34 (2009) 764–771


a single strain using POME as a substrate has not yet been studied in detail. Biohydrogen production can be achieved by...
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