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Novel Thermophilic Cellobiohydrolase






Researchers at the Joint BioEnergy Institute (JBEI) have developed a hyperthermophilic cellobiohydrolase protein (CBH) to break down cellulose into cellobiose at high temperatures. The JBEI CBH can be used in concert with JBEI endoglucanases (or any other endoglucanase with a similar temperature and ph profile) in high concentrations of ionic liquid. There is no loss of activity in ionic liquid concentrations up to 20%, and the enzyme cocktail is tolerant of ionic liquid concentrations of up to 50%. Therefore, the JBEI technology opens the possibility of one pot saccharification of lignocellulosic biomass. In addition, the JBEI technology is compatible, theoretically, with other pretreatment methods such as dilute acid and ammonia fiber explosion.

The JBEI CBH, isolated from Caldicellulosiruptor saccarolyticus, shows high CBH activity against insoluble substrates and has a higher tolerance to ionic liquid than acommercial enzyme cocktail developed for enzymatic hydrolysis of biomass. The enzyme retained more than 75% of the original activity over four days of incubation at 70ºC. Further, the technology functions at high temperatures (up to 80ºC), thus reducing the risk of contamination by other microorganisms

Current techniques treat biomass with a combination of high temperatures and an acid or base, or chemicals such as lime. One disadvantage of this method is that the industrial enzymes used to break down cellulose are not compatible with such harsh treatments. Therefore, single pot pretreatment/saccharification is impossible. In addition, current approaches create unwanted byproducts that interfere with the downstream hydrolysis and fermentation and do not lead to complete hydrolysis of the cellulose to sugars.

Ionic liquids have been shown to be effective in fully solubilizing lignocellulosic biomass. However, the addition of anti-solvents to separate the solubilized components of biomass (cellulose, hemicellulose and lignin) is an expensive and time consuming step. This new JBEI technology eliminates this step and maximizes hydrolysis.

EIO-2745 is compatible with JBEI invention EJIB-2666, Thermophilic Cellulases Compatible with Ionic Liquid Pretreatment.

The Joint BioEnergy Institute (JBEI, is a scientific partnership led by the Lawrence Berkeley National Laboratory and including the Sandia National Laboratories, the University of California campuses of Berkeley and Davis, the Carnegie Institution for Science and the Lawrence Livermore National Laboratory. JBEI’s primary scientific mission is to advance the development of the next generation of biofuels.


DEVELOPMENT STAGE: Proven principle.

STATUS: Patent pending. Available for licensing or collaborative research.


Fractionation and Removal of Solutes from Ionic Liquids, EJIB-2729

Recovery of Sugars by Solvent Extraction, EJIB-2750

Thermophilic Cellulases Compatible with Ionic Liquid Pretreatment, EJIB-2666

Thermophilic Endoglucanase Enzymes Engineered for Increased Activity, EIO-2667



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