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Enzyme-Catalyzed Hydrolysis of Cellulose in Ionic Liquids: A Green Approach Toward the Production of Biofuels

机译：离子液体中酶催化的纤维素水解：绿色途径生产生物燃料

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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 °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 °C in buffer while they are stable to temperatures as high as 115 °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.
机译：我们通过光学和量热技术研究了纤维素酶在八种离子液体中的商业混合物的反应性和稳定性。首先，将在这些离子液体中来自里氏木霉的纤维素酶的水解与在水性缓冲液中的水解进行基准。仅1-甲基咪唑氯化物（mim Cl）和三-（2-羟乙基）甲基硫酸甲基铵（HEMA）提供了可发生水解的介质。最初在缓冲液中于65°C水解要比在这两种液体中快得多，但在2小时后达到稳定水平，而在两种离子液体中反应则单调进行。水解速率的差异主要归因于两个因素：（1）离子液体的粘度较高；（2）酶在50°C的缓冲液中不可逆地变性，同时对高达115°C的温度稳定。 HEMA中的C。我们研究了酶的芳香族氨基酸的荧光猝灭是否确实是蛋白质变性的标志（如文献中所建议的），并得出结论，猝灭不一定与变性有关。当确实发生这种情况时，例如，在存在由咪唑阳离子和氯离子形成的离子液体的情况下，它是由咪唑而不是氯离子产生的。最后，我们得出结论，HEMA是进行纤维素水解反应以将生物质转化为生物燃料的一种有前途的新颖绿色介质。由于HEMA赋予酶的热稳定性，溶解生物质的能力以及不会淬灭色氨酸荧光的事实（因此允许通过荧光光谱法监控酶），因此HEMA为离子液体的设计提供了理想的起点不仅用于生物质的水解，而且还用于各种酶促反应。

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