Construction of bacterial artificial chromosome library from electrochemical microorganisms
Jung Ho Back
Biomedical Research Center, Korea Institute of Science and Technology, P. O. Box 131, Cheongryang, Seoul, Korea
The Graduate School of Biotechnology, Korea University, Seoul, Korea
Search for more papers by this authorMan Su Kim
Biomedical Research Center, Korea Institute of Science and Technology, P. O. Box 131, Cheongryang, Seoul, Korea
The Graduate School of Biotechnology, Korea University, Seoul, Korea
Search for more papers by this authorHyuk Cho
Biomedical Research Center, Korea Institute of Science and Technology, P. O. Box 131, Cheongryang, Seoul, Korea
Search for more papers by this authorIn Seop Chang
The Graduate School of Biotechnology, Korea University, Seoul, Korea
Search for more papers by this authorJiyoung Lee
The Graduate School of Biotechnology, Korea University, Seoul, Korea
Water Environment and Remediation Research Center, Korea Institute of Science and Technology, 39-1 Hawalkok-dong, Sungbuk-ku, Seoul 136-791, South Korea
Search for more papers by this authorKyung Sik Kim
The Graduate School of Biotechnology, Korea University, Seoul, Korea
Water Environment and Remediation Research Center, Korea Institute of Science and Technology, 39-1 Hawalkok-dong, Sungbuk-ku, Seoul 136-791, South Korea
Search for more papers by this authorByung Hong Kim
Water Environment and Remediation Research Center, Korea Institute of Science and Technology, 39-1 Hawalkok-dong, Sungbuk-ku, Seoul 136-791, South Korea
Search for more papers by this authorYoung In Park
The Graduate School of Biotechnology, Korea University, Seoul, Korea
Search for more papers by this authorCorresponding Author
Ye Sun Han
Department of Advanced Fusion Technology, Konkuk University, 1, Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea
*Corresponding author. Tel.: +82-2-958-5933; fax: +82-2-958-5909, E-mail address: [email protected]Search for more papers by this authorJung Ho Back
Biomedical Research Center, Korea Institute of Science and Technology, P. O. Box 131, Cheongryang, Seoul, Korea
The Graduate School of Biotechnology, Korea University, Seoul, Korea
Search for more papers by this authorMan Su Kim
Biomedical Research Center, Korea Institute of Science and Technology, P. O. Box 131, Cheongryang, Seoul, Korea
The Graduate School of Biotechnology, Korea University, Seoul, Korea
Search for more papers by this authorHyuk Cho
Biomedical Research Center, Korea Institute of Science and Technology, P. O. Box 131, Cheongryang, Seoul, Korea
Search for more papers by this authorIn Seop Chang
The Graduate School of Biotechnology, Korea University, Seoul, Korea
Search for more papers by this authorJiyoung Lee
The Graduate School of Biotechnology, Korea University, Seoul, Korea
Water Environment and Remediation Research Center, Korea Institute of Science and Technology, 39-1 Hawalkok-dong, Sungbuk-ku, Seoul 136-791, South Korea
Search for more papers by this authorKyung Sik Kim
The Graduate School of Biotechnology, Korea University, Seoul, Korea
Water Environment and Remediation Research Center, Korea Institute of Science and Technology, 39-1 Hawalkok-dong, Sungbuk-ku, Seoul 136-791, South Korea
Search for more papers by this authorByung Hong Kim
Water Environment and Remediation Research Center, Korea Institute of Science and Technology, 39-1 Hawalkok-dong, Sungbuk-ku, Seoul 136-791, South Korea
Search for more papers by this authorYoung In Park
The Graduate School of Biotechnology, Korea University, Seoul, Korea
Search for more papers by this authorCorresponding Author
Ye Sun Han
Department of Advanced Fusion Technology, Konkuk University, 1, Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea
*Corresponding author. Tel.: +82-2-958-5933; fax: +82-2-958-5909, E-mail address: [email protected]Search for more papers by this authorAbstract
A microbial fuel cell is a device that directly converts metabolic energy into electricity, using electrochemical technology. The analysis of large genome fragments recovered directly from microbial communities represents one promising approach to characterizing uncultivated electrochemical microorganisms. To further assess the utility of this approach, we constructed large-insert (140 kb) bacterial artificial chromosome (BAC) libraries from the genomic DNA of a microbial fuel cell, which had been operated for three weeks using acetate media. We screened the expression of several ferric reductase activities in the Escherichia coli host, in order to determine the extent of heterologous expression of metal-ion-reducing enzymes in the library. Phylogenetic analysis of 16S rRNA gene sequences recovered from the BAC libraries indicates that they contain DNA from a wide diversity of microbial organisms. The constructed bacterial library proved a powerful tool for exploring metal-ion reductase activities, providing information on the electron transport pathway of electrochemical microbial (ECM) organisms.
References
- [1] Lovely, D.R. (1991) Dissimilatory Fe(III) and Mn(IV) reduction. Microbiol. Rev. 55, 259–287.
- [2] Lovely, D.R., Giovannoni, S.J., White, D.C., Champine, J.E., Philips, E.J.P., Gorby, Y.A., Goodwin, S. (1993) Geobacter metallireducens gen. nov. sp. nov., a microorganism capable of coupling the complete oxidation of organic compounds to the reduction of iron and other metals. Arch. Microbiol. 159, 336–344.
- [3] Nealson, K.H., Saffarini, D. (1994) Iron and manganese in anaerobic respiration: environmental significance, physiology, and regulation. Ann. Rev. Microbiol. 48, 311–343.
- [4] Nealson, K.H., Myers, C. (1992) Microbial reduction of manganese and iron: new approaches to carbon cycling. Appl. Environ. Microbiol. 58, 439–443.
- [5] Myers, C.R., Myers, J.M. (1993) Ferric reductase is associated with the membranes of anaerobically grown Shewanella putrefaciens MR-1. FEMS Microbiol. Lett. 108, 15–22.
- [6] Myers, C.R., Myers, J.M. (1993) Role of menaquinone in the reduction of fumarate, nitrate, iron(III) and manganese(IV) by Shewanella putrefaciens MR-1. FEMS Microbiol. Lett. 114, 215–222.
- [7] Beliaev, A.S., Saffarini, D.A., Mclaughlin, J.L., Hunnicutt, D. (2001) MtrC, an outer membrane decahaem c cytochrome required for metal reduction in Shewanella putrefaciens MR-1. Mol. Microbiol. 39, 722–730.
- [8] Newman, D.K., Kolter, R. (2000) A role for excreted quinines in extracellular electron transfer. Nature 405, 94–97.
- [9] Seeliger, S., Cord-ruwisch, R., Schink, B. (1998) A periplasmic and extracellular c-type cytochrome of Geobacter sulfurreducens acts as a ferric iron reductase and as an electron carrier to other acceptors or to partner bacteria. J. Bacteriol. 180, 3686–3691.
- [10] Straub, K.L., Benz, M., Schink, B. (2001) Iron metabolism in anoxic environments at near neutral pH. FEMS Microbiol. Ecol. 34, 181–186.
- [11] Park, D.H., Zeikus, G. (2000) Electricity generation in microbial fuel cells using neutral red as an electronophore. Appl. Environ. Microbiol. 66, 1292–1297.
- [12] Coates, J.D., Ellis, D.J., Blunt-Harris, E.L., Gaw, C.V., Roden, E.E., Lovley, D.R. (1998) Recovery of humic-reducing bacteria from a diversity of environments. Appl. Environ. Microbiol. 64, 1504–1509.
- [13] Kim, H.J., Park, H.S., Hyun, M.S., Chang, I.S., Kim, M., Kim, B.H. (2002) A mediator-less microbial fuel cell using a metal reducing bacterium, Shewanella putrefaciens. Enzyme Microbiol. Technol. 30, 145–152.
- [14] Gil, G.C., Chang, I.S., Kim, M.A., Jang, J.K., Park, H.S., Kim, H.J., Kim, B.H. (2003) Operational parameters affecting the performance of a mediator-less microbial fuel cell. Biosens. Bioelectron. 18, 327–334.
- [15] Rondon, M.R., Raffel, S.J., Goodman, R.M., Handelsman, J. (1999) Toward functional genomics in bacteria: analysis of gene expression in Escherichia coli from a bacterial artificial chromosome library of Bacillus cereus. Proc. Natl. Acad. Sci. USA 96, 6451–6455.
- [16] Bürgmann, H., Pesaro, M., Widmer, F., Zeyer, J. (2001) A strategy for optimizing quality and quantity of DNA extracted from soil. J. Microbiol. Meth. 45, 7–20.
- [17] Dichrisitina, T.J., Delong, E.F. (1994) Isolation of anaerobic respiratory mutants of Shewanella Putrefaciens and genetic analysis of mutants deficient in anaerobic growth on Fe3+. J. Bacteriol. 176, 1468–1474.
- [18] Lawerence, Stookey L. (1970) Ferrozine - a new spectrophotometric reagent for iron. Anal. Chem. 42, 779–782.
- [19] Vartivarian, S.E., Cowart, R.E. (1999) Extracellular iron reductase: identification of a new class of enzymes by siderophore-producting microorganisms. Arch. Biochem. Biophys. 364, 75–82.
- [20] Park, H.S., Kim, B.H., Kim, H.S., Kim, H.J., Kim, G.T., Kim, M., Chang, I.S., Park, Y.K., Chang, H.I. (2001) A novel electrochemically active and Fe(III)-reducting bacterium phylogenetically related to clostridium butyricum isolated from a microbial fuel cell. Anaerobe 7, 297–306.
- [21] Lim, S.K., Park, D.H., Park, Y.K., Kim, B.H. (2001) Cloning, sequencing and functional expression in Escherichia coli of dmc gene encoding periplasmic tetraheme cytochrome C3 from Desulphovibrio desulphuricans M6. Anaerobe 7, 263–269.