Abstract
An anaerobic, extremely thermophilic, cellulolytic, non-spore-forming bacterium, strain 6A, was isolated from an alkaline hot spring in Hveragerði, Iceland. The bacterium was non-motile, rod-shaped (1.5–3.5x0.7 μm) and occurred singly, in pairs or in chains and stained gram-negative. The growth temperature was between 50 and 78°C with a temperature optimum near 68°C. Growth occurred between pH 5.8 and 8.2 with an optimum near 7.0. The bacterium fermented microcrystalline cellulose (Avicel) and produced lactate, acetate and H2 as the major fermentation products, and CO2 and ethanol occurred as minor fermentation products. Only a restricted number of carbon sources (cellulose, xylan, starch, pectin, cellobiose, xylose, maltose and lactose) were used as substrates. During growth on Avicel, the bacterium produced free cellulases with carboxymethylcellulase and avicelase activity. The G+C content of the cellular DNA of strain 6A was 35.2±0.8 mol%. Complete 16S rDNA sequence analysis showed that strain 6A was phylogenetically related to Caldicellulosiruptor saccharolyticus. It is proposed that the isolated bacterium be named Caldicellulosiruptor lactoaceticus sp. nov.
Similar content being viewed by others
References
Angelidaki I, Petersen SP, Ahring BK (1990) Effects of lipids on thermophilic anaerobic digestion and reduction of lipid inhibition upon addition of bentonite. Appl Microbiol Biotechnol 33:469–472
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Bryant MP (1972) Commentary on the Hungate technique for culture of anaerobic bacteria. Am J Clin Nutr 25:1324–1328
Cook GM, Janssen PH, Morgan HW (1991) Endospore formation by Thermoanaerobium brockii HTD4. Syst Appl Microbiol 14:240–244
Cord-Ruwisch R (1985) A quick method for the determination of dissolved and precipitated sulfides in cultures of sulfate-reducing bacteria. J Microbiol Methods 4:33–36
Dakhova ON, Kurepina NE, Zverlov VV, Svetlichnyi VA, Velikodvorskaya GA (1993) Cloning and expression in Escherichia coli of Thermotoga neapolitana genes coding for enzymes of carbohydrate substrate degradation. Biochem Biophys Res Commun 194:1359–1364
Gerhardt P (1981) Manual of methods for general bacteriology. American Society for Microbiology, Washington DC
Hudson JA, Morgan HW, Daniel RM (1990) A survey of cellulolytic anaerobic thermophiles from hot springs. Syst Appl Microbiol 13:72–76
Hudson JA, Morgan HW, Daniel RM (1991) The cellulase activity of an extreme thermophile. Appl Microbiol Biotechnol 35: 270–273
Hungate RE (1969) A role-tube method for cultivation of strict anaerobes. Methods Microbiol 3:117–132
Johnson EA, Sakajoh M, Halliwell G, Madia A, Demain AL (1982) Saccharification of complex cellulosic substrates by the cellulase system from Clostridium thermocellum. Appl Environ Microbiol 43:1125–1132
Mandels M, Andreotti R, Roche C (1976) Measurement of saccharifying cellulase. Biotech Bioeng Symp 6:21–33
Mathrani IM, Ahring BK (1992) Thermophilic and alkalophilic xylanases from several Dictyoglomus isolates. Appl Microbiol Biotechnol 38:23–27
Mathrani IM, Nielsen P, Sonne-Hansen J, Kristjánsson JK, Ahring BK (1993) Influence of pH and temperature on enumeration of cellulose-and hemicellulose-degrading thermophilic anaerobes in neutral and alkaline Icelandic hot springs. Appl Environ Microbiol 59:1963–1965
Mesbah M, Premachandran U, Whitman WB (1989) Precise measurement of the G+C content of deoxyribonucleic acid by high-performance liquid chromatography. Int J Syst Bacteriol 39:159–167
Miller GL (1959) Use of dinitrosalicylic acid as a reagent for the determination of reducing sugars. Exp Mycol 31:426–428
Nielsen P, Mathrani IM, Ahring BK (1993) Thermoanaerobium acetigenum spec. nov., a new anaerobic, extremely thermophilic, xylanolytic non-spore-forming bacterium isolated from an Icelandic hot spring. Arch Microbiol 159:460–464
Rainey FA, Ward NL, Morgan HW, Toalster R, Stackebrandt E (1993a) Phylogenetic analysis of anaerobic thermophilic bacteria: aid for their reclassification. J Bacteriol 175:4772–4779
Rainey FA, Janssen PH, Morgan HW, Stackebrandt E (1993b) A biphasic approach to the determination of the phenotypic and genotypic diversity of some anaerobic, cellulolytic, thermophilic, rod-shaped bacteria. Antonie Van Leeuwenhoek 64:341–355
Rainey FA, Donnison AM, Janssen PH, Saul D, Rodrigo A, Bergquist PL, Daniel RM, Stackebrandt E, Morgan HW (1994) Description of Caldicellulosiruptor saccharolyticus gen. nov., sp. nov: an obligately anaerobic, extremely thermophilic, cellulolytic bacterium. FEMS Microbiol Lett 120:263–266
Ruttersmith LD, Daniel RM (1991) Thermostable cellobiohydrolase from the thermophilic eubacterium Thermotoga sp. strain FjSS3-B.1. Biochem J 277:887–890
Schink B, Zeikus JG (1983) Clostridium thermosulfurogenes sp. nov., a new thermophile that produces elemental sulphur from thiosulphate. J Gen Microbiol 129:1149–1158
Sissons CH, Sharrock KR, Daniel RM, Morgan HW (1987) Isolation of cellulolytic anaerobic extreme thermophiles from New Zealand thermal sites. Appl Environ Microbiol 53:832–838
Sonne-Hansen J, Mathrani IM, Ahring BK (1993) Xylanolytic anaerobic thermophiles from Icelandic hot-springs. Appl Microbiol Biotechnol 38:537–541
Svetlichny VA, Svetlichnaya TP, Chernykh NA, Zavarzin GA (1990) Anaerocellum thermophilum gen. nov., sp.nov., an extreme thermophilic cellulolytic eubacterium isolated from hot springs in the Valley of Geysers. Mikrobiologiia 59:871–879
Taya M, Hinoki H, Yagi T, Kobayashi T (1988) Isolation and characterization of an extremely thermophilic, cellulolytic, anaerobic bacterium. Appl Microbiol Biotechnol 29:474–479
Wiegel J (1992) The obligately anaerobic thermophilic bacteria. In: Kristjansson JK (ed) Thermophilic bacteria. CRC Press, Boca Raton, pp 106–184
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Mladenovska, Z., Mathrani, I.M. & Ahring, B.K. Isolation and characterization of Caldicellulosiruptor lactoaceticus sp. nov., an extremely thermophilic, cellulolytic, anaerobic bacterium. Arch Microbiol 163, 223–230 (1995). https://doi.org/10.1007/BF00305357
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00305357