Enzyme-catalyzed hydrolysis of cellulose in ionic liquids: a green approach toward the production of biofuels.
We investigated the reactivity and stability of a commercial mixture of cellulases in eight ionic liquids by optical and calorimetric techniques. First, hydrolysis by cellulases from Tricoderma reesei in these ionic liquids was benchmarked against that in aqueous buffer. Only 1-methylimidazolium chloride (mim Cl) and tris-(2-hydroxyethyl)methylammonium methylsulfate (HEMA) provided a medium in which hydrolysis could occur. While hydrolysis at 65 degrees C is initially much faster in buffer than in these two liquids, it reaches a plateau after 2 h, whereas the reaction progresses monotonically in the two ionic liquids. This difference in the rate of hydrolysis is largely attributed to two factors: (1) the higher viscosity of the ionic liquids and (2) the enzymes are irreversibly denatured at 50 degrees C in buffer while they are stable to temperatures as high as 115 degrees C in HEMA. We explored whether fluorescence quenching of aromatic amino acids of the enzymes was indeed a signature of protein denaturation, as has been suggested in the literature, and concluded that quenching is not necessarily associated with denaturation. When it does occur, for example, in the presence of ionic liquids formed from imidazolium cations and chloride anions, it arises from the imidazolium rather than the chloride. Finally, we conclude that HEMA is a promising, novel, green medium for performing cellulose hydrolysis reactions to convert biomass into biofuels. Because of the thermal stability it imparts to enzymes, its ability to solubilize biomass, and the fact that it does not quench tryptophyl fluorescence (thus permitting monitoring of the enzymes by fluorescence spectroscopy), HEMA provides an ideal starting point for the design of ionic liquids, not only for the hydrolysis of biomass, but also for use with a wide spectrum of enzymatic reactions.
Department of Chemistry, Iowa State University, Ames, Iowa 50011, USA.
Sayantan
S
Bose
Daniel W
DW
Armstrong
Jacob W
JW
Petrich
D055162:Biocatalysis; D056804:Biofuels; D018533:Biomass; D044602:Cellulases / Q000378:metabolism*; D002482:Cellulose / Q000378:metabolism*; D006868:Hydrolysis; D052578:Ionic Liquids / Q000737:chemistry*; D055550:Protein Stability; D013696:Temperature; D014242:Trichoderma / Q000201:enzymology
D056804:Biofuels; D052578:Ionic Liquids; D002482:Cellulose; D044602:Cellulases