纤维素酶催化Aldol反应的研究  

Promiscuous Cellulase-Catalysed Aldol Reaction

作　　者：赖依峰[1,2] 葛新 罗能镇 相咸高 徐义明 章鹏飞 周俊超 冯烈 陈新志[1] 

机构地区：[1]浙江大学生物质化工教育部重点实验室,浙江大学化学工程与生物工程学系,浙江杭州310027 [2]浙江科技学院生物与化学工程学院,浙江杭州310012 [3]中建安装工程有限公司,江苏南京210046

出　　处：《高校化学工程学报》2013年第5期825-829,共5页Journal of Chemical Engineering of Chinese Universities

基　　金：高等学校博士学科点专项科研基金(20120101110062);浙江省低碳脂肪胺工程技术研究中心项目(2012E10033)资助

摘　　要：研究了纤维素酶在有机介质中多功能性催化芳香醛和丙酮的Aldol反应。考察了不同酶源对催化Aldol反应的影响,筛选出Cellulase from Trichoderma viride在DMSO中催化芳香醛和丙酮的Aldol反应的活性最高,与无酶空白对照实验和失活的Cellulase from Trichoderma viride的催化实验做了比较,未检测到产物的生成,确定纤维素酶可以催化苯甲醛和丙酮的Aldol反应。优化了反应溶剂、体系含水量、反应温度和酶浓度的工艺参数,在50℃,体系含水量为15%,酶浓度为10 mg·mL-1,在恒温振荡器中反应24 h,苯甲醛和丙酮的Aldol反应收率能达到85%。研究了各类的取代芳香醛与丙酮的Aldol反应情况,发现纤维素酶对于Aldol反应有着较强的底物适应性。同时提出了纤维素酶催化芳香醛和丙酮的Aldol反应的可能机理。It was observed that cellulase has a promiscuous ability of catalyzing Aldol reactions in the organic media. Six cellulases were screened according to their abilities of catalyzing the Aldol reaction between benzaldehyde and acetone. It was found that the enzyme of Cellulase from Trichoderma viride has the highest catalytic activity with using DMSO as the solvent, while using the denatured enzyme or the control experiment （in the absence of enzyme） demonstrates that no product of Aldol reaction can be detected. These results show that cellulase can catalyze Aldol reactions and also indicate that the enzymatic tertiary structure plays a critical role. The influence factors including solvents, temperature, water concent and enzyme concentration were optimized. When the Aldol reaction was conducted at 50℃ for 24 h under 15% water content and 10 mg-mL-1 enzyme concentration, the yield of 85% can be achieved. This paper studies the Aldol reaction between aromatic aldehydes and acetone. It was found that cellulase has a good adaptability for different substrates of the Aldol reaction and can produce high yield. The mechanism of the Aldol reaction catalyzed by cellulase was also proposed.

关 键 词：酶催化 纤维素酶 多功能性 ALDOL反应 

分 类 号：Q555.2[生物学—生物化学]