Abstract
A strictly anaerobic bacterium, strain B5T, was isolated from sediment of an abandoned coal mine in Taebaek, Republic of Korea. Cells of strain B5T were non-spore-forming, straight, Gram-positive rods. The optimum pH and temperature for growth were pH 7.0 and 30°C, respectively, while the strain was able to grow within pH and temperature ranges of 5.5–7.5 and 25–45°C, respectively. Growth of strain B5T was observed at NaCl concentrations of 0 to 6.0% (w/v) with an optimum at 3.0–4.0% (w/v). The polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol, an unknown phospholipid and three unknown polar lipids. Strain B5T grew anaerobically by reducing nitrate, nitrite, ferric-citrate, ferric-nitrilotriacetate, elemental sulfur, thiosulfate, and anthraquinone-2-sulfonate in the presence of proteinaceous compounds, organic acids, and carbohydrates as electron donors. The isolate was not able to grow by fermentation. Strain B5T did not grow under aerobic or microaerobic conditions. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain B5T is most closely related to the genus Tepidibacillus (T. fermentans STGHT; 96.3%) and Vulcanibacillus (V. modesticaldus BRT; 94.6%). The genomic DNA G+C content (36.9 mol%) of strain B5T was higher than those of T. fermentans STGHT (34.8 mol%) and V. modesticaldus BRT (34.5 mol%). Based on its phenotypic, chemotaxonomic, and phylogenetic properties, we describe a new species of a novel genus Calculibacillus, represented by strain B5T (=KCTC 15397T =JCM 19989T), for which we propose the name Calculibacillus koreensis gen. nov., sp. nov.
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Aklujkar, M., Coppi, M.V., Leang, C., Kim, B.C., Chavan, M.A., Perpetua, L.A., Giloteaux, L., Liu, A., and Holmes, D.E. 2013. Proteins involved in electron transfer to Fe(III) and Mn(IV) oxides by Geobacter sulfurreducens and Geobacter uraniireducens. Microbiology 159, 515–535.
Beller, H.R., Han, R., Karaoz, U., Lim, H., and Brodie, E.L. 2013. Genomic and physiological characterization of the chromatereducing, aquifer-derived Firmicute Pelosinus sp. strain HCF1. Appl. Environ. Microbiol. 79, 63–73.
Benson, H.J. 2002. Microbiological applications; a laboratory manual in general microbiology. 8th ed. McGraw Hill, New York, USA.
Blodau, C., Hoffmann, S., Peine, A., and Peiffer, S. 1998. Iron and sulfate reduction in the sediments of acidic mine lake 116 (Brandenburg, Germany): Rates and geochemical evaluation. Water Air Soil Pollut. 108, 249–270.
Buck, J.D. 1982. Nonstaining (KOH) method for determination of gram reactions of marine bacteria. Appl. Environ. Microbiol. 44, 992–993.
Caccavo, F., Blakemore, R.P., and Lovley, D.R. 1992. A hydrogenoxidizing, Fe(III)-reducing microorganism from the Great Bay estuary, New Hampshire. Appl. Environ. Microbiol. 58, 3211–3216.
Caccavo, F. Jr., Lonergan, D.J., Lovley, D.R., Davis, M., Stolz, J.F., and McInerney, M.J. 1994. Geobacter sulfurreducens sp. nov., a hydrogen- and acetate-oxidizing dissimilatory metal-reducing microorganism. Appl. Environ. Microbiol. 60, 3752–3759.
Childers, S.E., Ciufo, S., and Lovley, D.R. 2002. Geobacter metallireducens accesses insoluble Fe(III) oxide by chemotaxis. Nature 416, 767–769.
Felsenstein, J. 1981. Evolutionary trees from DNA sequences: A maximum likelihood approach. J. Mol. Evol. 17, 368–376.
Friese, K., Wendt-Potthoff, K., Zachmann, D.W., Fauville, A., Mayer, B., and Veizer, J. 1998. Biogeochemistry of iron and sulfur in sediments of an acidic mining lake in Lusatia, Germany. Water Air Soil Pollut. 108, 231–247.
Gonzalez, J.M. and Saiz-Jimenez, C. 2002. A fluorimetric method for the estimation of G+C mol% content in microorganisms by thermal denaturation temperature. Environ. Microbiol. 4, 770–773.
Hall, T.A. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl. Acids Symp. Ser. 41, 95–98.
Hong, Y., Wu, P., Li, W., Gu, J., and Duan, S. 2012. Humic analog AQDS and AQS as an electron mediator can enhance chromate reduction by Bacillus sp. strain 3C3. Appl. Microbiol. Biotechnol. 93, 2661–2668.
Islam, F.S., Gault, A.G., Boothman, C., Polya, D.A., Charnock, J.M., Chatterjee, D., and Lloyd, J.R. 2004. Role of metal-reducing bacteria in arsenic release from Bengal delta sediments. Nature 430, 68–71.
Johnson, D.B., Rolfe, S., Hallberg, K.B., and Iversen, E. 2001. Isolation and phylogenetic characterization of acidophilic microorganisms indigenous to acidic drainage waters at an abandoned Norwegian copper mine. Environ. Microbiol. 3, 630–637.
Kaneda, T. 1991. Iso-and anteiso-fatty acids in bacteria: Biosynthesis, function, and taxonomic significance. Microbiol. Rev. 55, 288–302.
Kim, O.S., Cho, Y.J., Lee, K., Yoon, S.H., Kim, M., Na, H., Park, S.C., Jeon, Y. S., Lee, J.H., Yi, H., et al. 2012. Introducing EzTaxone-e: a prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species. Int. J. Syst. Evol. Microbiol. 62, 716–721.
Kim, K.K., Lee, J.S., Lee, K.C., Oh, H.M., and Kim, S.G. 2010. Pontibaca methylaminivorans gen. nov., sp. nov., a member of the family Rhodobacteraceae. Int. J. Syst. Evol. Microbiol. 60, 2170–2175.
Kimura, M. 1983. The neutral theory of molecular evolution. Cambridge University Press, Cambridge.
L’Haridon, S., Miroshnichenko, M.L., Kostrikina, N.A., Tindall, B.J., Spring, S., Schumann, P., Stackebrandt, E., Bonch-Osmolovskaya, E.A., and Jeanthon, C. 2006. Vulcanibacillus modesticaldus gen. nov., sp. nov., a strictly anaerobic, nitrate-reducing bacterium from deep-sea hydrothermal vents. Int. J. Syst. Evol. Microbiol. 56, 1047–1053.
Lane, D. 1991. 16S/23S rRNA sequencing, pp. 115–175. In Stackebrandt, E. and Goodfellow, M. (eds.), Nucleic acid techniques in bacterial systematics, Wiley, Chichester, UK.
Li, X., Park, J.H., Edraki, M., and Baumgartl, T. 2013. Understanding the salinity issue of coal mine spoils in the context of salt cycle. Environ. Geochem. Health 36, 453–465.
Li, H., Peng, J., Weber, K.A., and Zhu, Y. 2011. Phylogenetic diversity of Fe(III)-reducing microorganisms in rice paddy soil: Enrichment cultures with different short-chain fatty acids as electron donors. J. Soils Sediments 11, 1234–1242.
Lovley, D.R. 1991. Dissimilatory Fe(III) and Mn(IV) reduction. Microbiol. Rev. 55, 259–287.
Lovley, D.R., Chapelle, F.H., and Phillips, E.J. 1990. Fe(III)-reducing bacteria in deeply buried sediments of the Atlantic Coastal Plain. Geology 18, 954–957.
Lovley, D.R. and Phillips, E.J. 1986. Organic matter mineralization with reduction of ferric iron in anaerobic sediments. Appl. Environ. Microbiol. 51, 683–689.
Lovley, D.R. and Phillips, E.J. 1988. Novel mode of microbial energy metabolism: Organic carbon oxidation coupled to dissimilatory reduction of iron or manganese. Appl. Environ. Microbiol. 54, 1472–1480.
Meslé, M., Dromart, G., and Oger, P. 2013. Microbial methanogenesis in subsurface oil and coal. Res. Microbiol. 164, 959–972.
Minnikin, D.E., O’Donnell, A.G., Goodfellow, M., Alderson, G., Athalye, M., Schaal, A., and Parlett, J.H. 1984. An integrated procedure for the extraction of bacterial isoprenoid quinones and polar lipids. J. Microbiol. Methods 2, 233–241.
Nei, M., Kumar, S., and Takahashi, K. 1998. The optimization principle in phylogenetic analysis tends to give incorrect topologies when the number of nucleotides or amino acids used is small. Proc. Natl. Acad. Sci. USA 95, 12390–12397.
Nevin, K.P., Holmes, D.E., Woodard, T.L., Hinlein, E.S., Ostendorf, D.W., and Lovley, D.R. 2005. Geobacter bemidjiensis sp. nov. and Geobacter psychrophilus sp. nov., two novel Fe(III)-reducing subsurface isolates. Int. J. Syst. Evol. Microbiol. 55, 1667–1674.
Ogg, C.D. and Patel, B.K. 2009. Thermotalea metallivorans gen. nov., sp. nov., a thermophilic, anaerobic bacterium from the Great Artesian Basin of Australia aquifer. Int. J. Syst. Evol. Microbiol. 59, 964–971.
Pruesse, E., Quast, C., Knittel, K., Fuchs, B.M., Ludwig, W., Peplies, J., and Glockner, F.O. 2007. SILVA: a comprehensive online resource for quality checked and aligned ribosomal RNA sequence data compatible with ARB. Nucleic Acids Res. 35, 7188–7196.
Saitou, N. and Nei, M. 1987. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol. Biol. Evol. 4, 406–425.
Sasser, M. 1990. Identification of bacteria by gas chromatography of cellular fatty acids. MIDI Technical Note 101., MIDI Inc, Newark, DE,USA.
Schaeffer, A.B. and Fulton, M.D. 1933. A simplified method of staining endospores. Science 77, 194.
Semple, K., Westlake, D., and Krouse, H. 1987. Sulfur isotope fractionation by strains of Alteromonas putrefaciens isolated from oil field fluids. Can. J. Microbiol. 33, 372–376.
Shirling, E.B. and Gottlieb, D. 1966. Methods for characterization of Streptomyces species. Int. J. Syst. Evol. Microbiol. 16, 313–340.
Si, O.J., Kim, S.J., Jung, M.Y., Choi, S.B., Kim, J.G., Kim, S.G., Roh, S.W., Lee, S., and Rhee, S.K. 2015. Leeuwenhoekiella polynyae sp. nov., isolated from a polynya in western Antarctica. Int. J. Syst. Evol. Microbiol. 65, 1694–1699.
Siegert, M., Cichocka, D., Herrmann, S., Grundger, F., Feisthauer, S., Richnow, H.H., Springael, D., and Krüger, M. 2011. Accelerated methanogenesis from aliphatic and aromatic hydrocarbons under iron- and sulfate-reducing conditions. FEMS Microbiol. Lett. 315, 6–16.
Slobodkin, A., Reysenbach, A.L., Strutz, N., Dreier, M., and Wiegel, J. 1997. Thermoterrabacterium ferrireducens gen. nov., sp. nov., a thermophilic anaerobic dissimilatory Fe(III)-reducing bacterium from a continental hot spring. Int. J. Syst. Bacteriol. 47, 541–547.
Slobodkina, G.B., Panteleeva, A.N., Kostrikina, N.A., Kopitsyn, D.S., Bonch-Osmolovskaya, E.A., and Slobodkin, A.I. 2013. Tepidibacillus fermentans gen. nov., sp. nov.: A moderately thermophilic anaerobic and microaerophilic bacterium from an underground gas storage. Extremophiles 17, 833–839.
Sororzano, L. 1969. Determination of ammonia in natural waters by the phenolhypochlorite method. Limnol. Oceanogr. 14, 799–801.
Tamura, K., Stecher, G., Peterson, D., Filipski, A., and Kumar, S. 2013. MEGA6: Molecular evolutionary genetics analysis version 6.0. Mol. Biol. Evol. 30, 2725–2729.
Tan, B., Charchuk, R., Li, C., Nesbo, C., Abu Laban, N., and Foght, J. 2014. Draft genome sequence of uncultivated Firmicutes (Peptococcaceae SCADC) single cells sorted from methanogenic alkane-degrading cultures. Genome Announc. 2, e00909–14.
Trüper, H.G. and Schlegel, H.G. 1964. Sulphur metabolism in Thiorhodaceae. I. Quantitative measurements on growing cells of Chromatium okenii. Antonie van Leeuwenhoek 30, 225–238.
Tschech, A. and Pfennig, N. 1984. Growth yield increase linked to caffeate reduction in Acetobacterium woodii. Arch. Microbiol. 137, 163–167.
Weber, K.A., Achenbach, L.A., and Coates, J.D. 2006. Microorganisms pumping iron: anaerobic microbial iron oxidation and reduction. Nat. Rev. Microbiol. 4, 752–764.
Weisburg, W.G., Barns, S.M., Pelletier, D.A., and Lane, D.J. 1991. 16S ribosomal DNA amplification for phylogenetic study. J. Bacteriol. 173, 697–703.
Widdel, F. and Bak, F. 1992. Gram-negative mesophilic sulfate-reducing bacteria, pp. 3352–3378. In Starr, M.P., Stolp, H., Trüper H.G., Balows, A., and Schlegal, H.G. (eds.), The Prokaryotes, 2nd ed vol. 1. Springer, Berlin, Germany.
Wrighton, K.C., Agbo, P., Warnecke, F., Weber, K.A., Brodie, E.L., DeSantis, T.Z., Hugenholtz, P., Andersen, G.L., and Coates, J.D. 2008. A novel ecological role of the Firmicutes identified in thermophilic microbial fuel cells. ISME J. 2, 1146–1156.
Yarza, P., Yilmaz, P., Pruesse, E., Glöckner, F.O., Ludwig, W., Schleifer, K.H., Whitman, W.B., Euzéby, J., Amann, R., and Rosselló-Móra, R. 2014. Uniting the classification of cultured and uncultured bacteria and archaea using 16S rRNA gene sequences. Nat. Rev. Microbiol. 12, 635–645.
Zhang, Y.Z., Fang, M.X., Zhang, W.W., Li, T.T., Wu, M., and Zhu, X.F. 2013. Salimesophilobacter vulgaris gen. nov., sp. nov., an anaerobic bacterium isolated from paper-mill wastewater. Int. J. Syst. Evol. Microbiol. 63, 1317–1322.
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Min, UG., Kim, SJ., Hong, H. et al. Calculibacillus koreensis gen. nov., sp. nov., an anaerobic Fe(III)-reducing bacterium isolated from sediment of mine tailings. J Microbiol. 54, 413–419 (2016). https://doi.org/10.1007/s12275-016-6086-8
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DOI: https://doi.org/10.1007/s12275-016-6086-8