Stable cellulases are critical for in situ hydrolysis of ionic liquid-pretreated lignocellulose, and surfactants were reported to improve the stability of cellulase in practical circumstances. In this study, synergistic effect of Tween-80 and [Emim]OAc was first identified to enhance the hydrolysis of lignocellulose by Paenibacillus sp. LLZ1 cellulase, proving the effectiveness of surfactants in in situ hydrolysis. Protein fluorescence and near-UV circular dichroism spectroscopy analyses illustrated that ionic liquids might move local residues and alter global tertiary structure of cellulase, leading to higher instability. The addition of Tween-80 might turn the microenvironment of cellulase residues more hydrophilic and reduce the contact of protein with deactivation factors. The synergistic system was further applied in the in situ hydrolysis of microcrystalline cellulose and bagasse cellulose by Paenibacillus cellulase. The enzymatic hydrolysis rate of microcrystalline cellulose was enhanced by 2.5-fold and the final conversion was improved by 45%. Scanning electron microscopy demonstrated the substrate solubilizing effect during reaction by monitoring the morphological changes of lignocellulosic substrates. This study highlighted the potential of applying synergistic reaction system in in situ enzymatic hydrolysis of lignocellulose, and revealed the inherent mechanism initially.

稳定的纤维素酶对于离子液体预处理的木质纤维素的原位水解至关重要，据报道，在实际情况下，表面活性剂可改善纤维素酶的稳定性。在这项研究中，首先确定了Tween-80和[Emim] OAc的协同作用可增强Paenibacillus sp。对木质纤维素的水解作用。LLZ1纤维素酶，证明表面活性剂的原位有效性水解。蛋白质荧光和近紫外圆二色性光谱分析表明，离子液体可能会移动纤维素酶的局部残留并改变纤维素酶的整体三级结构，从而导致更高的不稳定性。添加Tween-80可能会使纤维素酶残基的微环境更亲水，并减少蛋白质与失活因子的接触。该协同系统进一步应用于Paenibacillus原位水解微晶纤维素和蔗渣纤维素纤维素酶。微晶纤维素的酶水解速率提高了2.5倍，最终转化率提高了45％。扫描电子显微镜通过监测木质纤维素底物的形态变化证明了反应过程中底物的溶解作用。这项研究突出了在木质纤维素原位酶水解中应用协同反应系统的潜力，并初步揭示了其内在机理。