Expression of Acetaldehyde Dehydrogenase Gene Increases Hydrogen Production and Acetate Consumption by Rhodobacter sphaeroides

Shinya Hasegawa; Jyumpei Kobayashi; Tomoe Komoriya; Hideki Kohno; Kazuaki Yoshimune

doi:10.4236/epe.2015.79037

Energy and Power Engineering > Vol.7 No.9, August 2015

Expression of Acetaldehyde Dehydrogenase Gene Increases Hydrogen Production and Acetate Consumption by Rhodobacter sphaeroides ()

Shinya Hasegawa¹, Jyumpei Kobayashi², Tomoe Komoriya³, Hideki Kohno^1,3, Kazuaki Yoshimune¹

¹Department of Applied Molecular Chemistry, Graduate School of Industrial Technology, Nihon University, Narashino, Chiba, Japan.
²Microbial Genetic Division, Institute of Genetic Resources, Faculty of Agriculture, Kyushu University, Fukuoka, Japan.
³Department of Sustainable Engineering, College of Industrial Technology, Nihon University, Narashino, Chiba, Japan.

DOI: 10.4236/epe.2015.79037 PDF HTML XML 2,741 Downloads 3,154 Views

Abstract

Rhodobacter sphaeroides RV (RV) produces high yields of hydrogen from organic acids in the presence of light. The hydrogen production from acetate is lower than that from lactate, probably because of its low ability to metabolize acetate. In this study, gene of acetaldehyde dehydrogenase (ACDH, EC 1.2.1.10) that catalyzes the reversible conversion of acetaldehyde and CoA to acetyl-CoA with the concurrent reduction of NAD to NADH, is overexpressed in the RV strain. The produced acetyl-CoA can be oxidized to carbon dioxide in the tricarboxylic acid (TCA) cycle, wherein electrons are generated and used for hydrogen production. The byproduct NADH can be used as reducing agent for acetate to produce acetaldehyde by acetate dehydrogenase. The recombinant RV strain (RVAC) expressing the ACDH gene showed ACDH activity with a specific activity of 3.2 mU/ mg, and the RV and the recombinant RV strain that harbored the intact (empty) plasmid pLP-1.2 (RVI) showed no detectable ACDH activity. The hydrogen yields of the RVAC strain from 21-mM acetate were 1.5-fold higher than that of the wild type RV strain and also that of the RVI strain. In contrast, hydrogen yield from 21-mM lactate was 30% lower than that in the control strains.

Keywords

Acetate, Lactate, Photosynthetic Bacteria, Rhodobacter sphaeroides, Aldehyde Dehydrogenase

Share and Cite:

Hasegawa, S. , Kobayashi, J. , Komoriya, T. , Kohno, H. and Yoshimune, K. (2015) Expression of Acetaldehyde Dehydrogenase Gene Increases Hydrogen Production and Acetate Consumption by Rhodobacter sphaeroides. Energy and Power Engineering, 7, 396-402. doi: 10.4236/epe.2015.79037.

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Das, D. (2009) Advances in Biohydrogen Production Processes: An Approach Towards Commercialization. International Journal of Hydrogen Energy, 34, 7349-7357. http://dx.doi.org/10.1016/j.ijhydene.2008.12.013
[2]	Sun, Q., Xiao, W., Xi, D., Shi, J., Yan, X. and Zhou, Z. (2010) Statistical Optimization of Biohydrogen Production from Sucrose by a Co-Culture of Clostridium acidisoli and Rhodobacter sphaeroides. International Journal of Hydrogen Energy, 35, 4076-4084. http://dx.doi.org/10.1016/j.ijhydene.2010.01.145
[3]	Nath, K., Muthukumar, M., Kumar, A. and Das, D. (2008) Kinetics of Two-Stage Fermentation Process for the Production of Hydrogen. International Journal of Hydrogen Energy, 33, 1195-1203. http://dx.doi.org/10.1016/j.ijhydene.2007.12.011
[4]	Fang, H.H.P., Zhu, H. and 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. http://dx.doi.org/10.1016/j.ijhydene.2006.03.005
[5]	Redwood, M.D. and Macaskie, L.E. (2006) A Two-Stage, Two-Organism Process for Biohydrogen from Glucose. International Journal of Hydrogen Energy, 31, 1514-1521. http://dx.doi.org/10.1016/j.ijhydene.2006.06.018
[6]	Asada, Y., Ohsawa, M., Nagai, Y., Ishimi, K., Fukatsu, M., Hideno, A., Wakayama, T. and Miyake, J. (2008) Re-Eva-luation of Hydrogen Productivity from Acetate by Some Photosynthetic Bacteria. International Journal of Hydrogen Energy, 33, 5147-5150. http://dx.doi.org/10.1016/j.ijhydene.2008.05.005
[7]	Kobayashi, J., Hasegawa, S., Itou, K., Yoshimune, K., Komoriya, K., Asada, Y. and Kohno, H. (2012) Expression of Aldehyde Dehydrogenase Gene Increases Hydrogen Production from Low Concentration of Acetate by Rhodobacter sphaeroides. International Journal of Hydrogen Energy, 37, 9602-9609. http://dx.doi.org/10.1016/j.ijhydene.2012.03.053
[8]	Barbosa, M.J., Rocha, J.M., Tramper, J. and Wijffels, R.H. (2001) Acetate as a Carbon Source for Hydrogen Production by Photosynthetic Bacteria. Journal of Biotechnology, 85, 25-33. http://dx.doi.org/10.1016/S0168-1656(00)00368-0
[9]	Wakayama, T., Nakada, E., Asada, Y. and Miyake, J. (2000) Effect of Light/Dark Cycle on Bacterial Hydrogen Production by Rhodobacter sphaeroides RV. Applied Biochemistry and Biotechnology, 84–86, 431-440. http://dx.doi.org/10.1385/ABAB:84-86:1-9:431
[10]	Wakayama, T. and Miyake, J. (2002) Light Shade Bands for the Improvement of Solar Hydrogen Production Efficiency by Rhodobacter sphaeroides RV. International Journal of Hydrogen Energy, 27, 1495-500. http://dx.doi.org/10.1016/S0360-3199(02)00088-5
[11]	Kobayashi, J., Yoshimune, K., Komoriya, T. and Kohno, H. (2011) Efficient Hydrogen Production from Acetate through Isolated Rhodobacter sphaeroides. Journal of Bioscience and Bioengineering, 112, 602-605. http://dx.doi.org/10.1016/j.jbiosc.2011.08.008
[12]	Klyosov, A.A., Rashkovetsky, L.G., Tahir, M.K. and Keung, M.M. (1996) Possible Role of Liver Cytosolic and Mitochondrial Aldehyde Dehydrogenase in Acetaldehyde Metabolism. Biochemistry, 35, 4445-4456. http://dx.doi.org/10.1021/bi9521093
[13]	Goldberg, R.N., Tewari, Y.B., Bell, D. and Fazio, K. (1993) Thermodynamics of Enzyme-Catalyzed Reactions: Part 1. Oxidoreductases. Journal of Physical and Chemical Reference Data, 22, 515-582. http://dx.doi.org/10.1063/1.555939
[14]	Rudolph, F.B., Purich, D.L. and Fromm, H.J. (1968) Coenzyme A-Linked Aldehyde Dehydrogenase from Escherichia coli. I. Partial Purification, Properties, and Kinetic Studies of the Enzyme. The Journal of Biological Chemistry, 243, 5539-5545.
[15]	Rupprecht, J., Hankamer, B., Mussgnug, J.H., Ananyev, G., Dismukes, C. and Kruse, O. (2006) Perspectives and Advances of Biological H2 Production in Microorganisms. Applied Microbiology and Biotechnology, 72, 442-449. http://dx.doi.org/10.1007/s00253-006-0528-x
[16]	Koku, H., Eroglu, I., Gündüz, U., Yücel, M. and Türker, L. (2002) Aspects of the Metabolism of Hydrogen Production by Rhodobacter sphaeroides. International Journal of Hydrogen Energy, 27, 1315-1329. http://dx.doi.org/10.1016/S0360-3199(02)00127-1
[17]	Kanehisa, M., Goto, S., Sato, Y., Furumichi, M. and Tanabe, M. (2012) KEGG for Integration and Interpretation of Large-Scale Molecular Datasets. Nucleic Acids Research, 40, D109-D114. http://dx.doi.org/10.1093/nar/gkr988
[18]	Kanehisa, M. and Goto, S. (2000) KEGG: Kyoto Encyclopedia of Genes and Genomes. Nucleic Acids Research, 28, 27-30. http://dx.doi.org/10.1093/nar/28.1.27
[19]	Lee, S.J., Ko, J.H., Kang, H.Y. and Lee, Y. (2006) Coupled Expression of MhpE Aldolase and MhpF Dehydrogenase in Escherichia coli. Biochemical and Biophysical Research Communications. 346, 1009-1015. http://dx.doi.org/10.1016/j.bbrc.2006.06.009
[20]	Ferrández, A., Garciá, J.L. and Díaz, E. (1997) Genetic Characterization and Expression in Heterologous Hosts of the 3-(3-Hydroxyphenyl)propionate Catabolic Pathway of Escherichia coli K-12. Journal of Bacteriology, 179, 2573-2581.