纤维素酶的低共熔溶剂双水相萃取研究

全文下载： 202303003.pdf

文章编号： 1672-6987（2023）03-0023-07； DOI： 10.16351/j.1672-6987.2023.03.003

付玉洁， 于海龙*(青岛科技大学 化工学院，山东 青岛 266042)

摘要： 为寻找新型绿色溶剂体系改善酶的活性和可回收性，本研究将低共熔溶剂双水相体系应用于纤维素酶的回收。为此合成了不同氯化胆碱（ChCl）基低共熔溶剂改善纤维素酶活性并构建双水相体系回收纤维素酶。将ChCl-聚乙二醇作为萃取溶剂，通过单因素实验考察了DES用量、盐浓度、纤维素酶质量、萃取时间和温度等因素对萃取效果的影响。实验结果表明，在优化的提取条件下（当双水相体系中K2HPO4浓度为025 g·mL-1，DES体积分数35%，在25 ℃下对较少纤维素酶萃取30 min时），可以从水解液中将90%左右的酶萃取到上相。通过对纤维素酶的紫外光谱和荧光光谱的研究发现，萃取过程中纤维素酶的构象没有发生变化。利用电导率和动态光散射（DLS）研究了萃取机理，结果表明：DES聚集体的形成在萃取过程中发挥了重要作用。所有结果表明，以低共熔溶剂为基础的双水相体系在纤维素酶回收方面提供了新的可能性。

关键词： 低共熔溶剂； 双水相体系； 萃取； 纤维素酶

中图分类号： TQ 413文献标志码： A

引用格式： 付玉洁， 于海龙. 纤维素酶的低共熔溶剂双水相萃取研究［J］. 青岛科技大学学报(自然科学版), 2023, 44(3): 23-29.

FU Yujie， YU Hailong. Deep eutectic solvent aqueous two-phase system for cellulase extracting ［J］. Journal of Qingdao University of Science and Technology（Natural Science Edition), 2023， 44（3）： 23-29.

Deep Eutectic Solvent Aqueous Two-phase System for Cellulase Extracting

FU Yujie， YU Hailong

(College of Chemical Engineering, Qingdao University of Science and Technology, Qingdao 266042, China)

Abstract: In order to find a new green solvent system to improve the activity and recyclability of cellulase, a deep eutectic solvent two-phase system was applied to the recovery of cellulase for the first time in this paper. Different choline chloride (ChCl)-based DES were synthesized to improve cellulase activity and constructed a two-phase system to recover cellulase. ChCl-polyethylene glycol was selected as the suitable extraction solvent，and single factor experiments have been done to investigate the effects of the extraction process, including the amount of DES, the concentration of salt, the mass of cellulase, the extraction time and the temperature. The experimental results show that under the optimized extraction conditions（When the K2HPO4 concentration in the dual-water phase system is 025 g·mL-1, the contont of the DES is 35%, when the less cellulose enzyme is extracted for 30 min at 25 ℃）, about 90% of the enzymes can be extracted from the hydrolysate to the upper phase. By studying the UV and fluorescence spectra of cellulase, it was found that the conformation of cellulase did not change during the extraction process; and the extraction mechanism was studied by electrical conductivity and dynamic light scattering (DLS). The results indicated that the formation of DES aggregates plays an important role in the extraction process. All results suggest that deep eutectic solvent-based aqueous two-phase systems offer new possibilities in cellulase recovery.

Key words: deep eutectic solvent; aqueous two-phase system; extraction; cellulase

收稿日期： 2022-04-24

基金项目： 国家自然科学基金项目（32171735）；山东省自然科学基金项目（ZR2019BC043）.

作者简介： 付玉洁 (1996—), 男, 硕士研究生.*通信联系人.