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Background  Consolidated bioprocessing (CBP) of lignocellulosic biomass to ethanol using thermophilic bacteria provides a promising solution for efficient lignocellulose conversion without the need for additional cellulolytic enzymes. Most studies on the thermophilic CBP concentrate on co-cultivation of the thermophilic cellulolytic bacterium  Clostridium thermocellum  with non-cellulolytic thermophilic anaerobes at temperatures of 55°C-60°C.    Results  We have specifically screened for cellulolytic bacteria growing at temperatures >70°C to enable direct conversion of lignocellulosic materials into ethanol. Seven new strains of extremely thermophilic anaerobic cellulolytic bacteria of the genus  Caldicellulosiruptor  and eight new strains of extremely thermophilic xylanolytic/saccharolytic bacteria of the genus  Thermoanaerobacter  isolated from environmental samples exhibited fast growth at 72°C, extensive lignocellulose degradation and high yield ethanol production on cellulose and pretreated lignocellulosic biomass. Monocultures of  Caldicellulosiruptor  strains degraded up to 89-97% of the cellulose and hemicellulose polymers in pretreated biomass and produced up to 72 mM ethanol on cellulose without addition of exogenous enzymes. In dual co-cultures of  Caldicellulosiruptor  strains with  Thermoanaerobacter  strains the ethanol concentrations rose 2- to 8.2-fold compared to cellulolytic monocultures. A co-culture of  Caldicellulosiruptor  DIB 087C and  Thermoanaerobacter  DIB 097X was particularly effective in the conversion of cellulose to ethanol, ethanol comprising 34.8 mol% of the total organic products. In contrast, a co-culture of  Caldicellulosiruptor saccharolyticus  DSM 8903 and  Thermoanaerobacter mathranii  subsp.  mathranii  DSM 11426 produced only low amounts of ethanol.    Conclusions  The newly discovered  Caldicellulosiruptor  sp. strain DIB 004C was capable of producing unexpectedly large amounts of ethanol from lignocellulose in fermentors. The established co-cultures of new  Caldicellulosiruptor  strains with new  Thermoanaerobacter  strains underline the importance of using specific strain combinations for high ethanol yields. These co-cultures provide an efficient CBP pathway for ethanol production and represent an ideal starting point for development of a highly integrated commercial ethanol production process.

Single-step ethanol production from lignocellulose using novel extremely thermophilic bacteria