三种侧耳漆酶在烷基咪唑类离子液体中催化特性的研究  

Catalytic Properties of Three Pleurotus sp. Laccases in Alkylimidazolium Based Ionic Liquids

作　　者：孙见良 伍星[1] 刘浩[1] 刘明[2] 何焕清[2] 许向阳 Sun fian-liang Wu Xing Liu Hao Liu Ming He Huan-qing Xu Xiang-yang(College of Light Industry Science and Engineering , South China University of Technology, Guangzhou 510640, Guangdong, China Vegetable Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, Guangdong, China Zaozhuang Jienuo Enzyme Co. Ltd. , Zaozhuang 277100, Shandong, China)

机构地区：[1]华南理工大学轻工科学与工程学院,广东广州510640 [2]广东省农业科学院蔬菜研究所,广东广州510640 [3]枣庄市杰诺生物酶有限公司,山东枣庄277100

出　　处：《造纸科学与技术》2016年第5期48-53,共6页Paper Science & Technology

基　　金：国家自然科学基金项目(31570569);制浆造纸工程国家重点实验室项目(2015QN03)

摘　　要：漆酶在纸浆漂白、纤维改性、废纸脱墨、废水氯代酚降解等方面有广阔的应用前景,使用离子液体/水混合溶剂可以提高漆酶催化不溶性底物合成或降解的效率,但同时会造成酶的活力下降。作者从红平菇、榆黄菇和糙皮侧耳菌糠中分离得到漆酶并研究了它们在烷基咪唑类离子液体中的催化特性。结果表明,红平菇、榆黄菇和糙皮侧耳漆酶在三种离子液体中均受到抑制作用。几种溶剂的抑制作用排序如下:乙醇<丙酮<1-乙基-3-甲基咪唑硫酸乙酯盐([EMIM]EtOSO_3)<1-丁基-3-甲基咪唑磷酸二丁酯([BMIM]DBP)<1-乙基-3-甲基咪唑醋酸盐([EMIM]Ac)。动力学分析表示,离子液体[EMIM]EtOSO_3与底物2,2'-连氮-二(3-乙基苯并噻唑啉-6-磺酸)(ABTS)对酶分子活性位点发生了竞争性结合,因此推测,漆酶在离子液体中的表观活力与初始酶用量无关,增加ABTS浓度有利于提高反应速率和酶的利用效率。Laccase has broad application prospects in pulp biobleaching,fiber modification,wastepaper deinking and degradation of chlorophenols in pulping effluents. Using water- ionic liquid（ IL） mixed solvents could improve the catalytic efficiency of laccase in synthesis or degradation of water- insoluble substrates. However,the presence of ILs would also denature laccase. In this work,laccases were isolated from the mushroom culture waste of Pleurotus djamor,Pleurotus citrinopileatus and Pleurotus ostreatus. Their catalytic properties in alkylimidazolium based ILs were studied. Results showed all the isolated Pleurotus sp. laccases were strongly inhibited by the alkylimidazolium ILs. The inhibitory effects of organic solvents followed an order： ethanol 〈 acetone 〈 [EMIM]Et OSO3〈 [BMIM]DBP 〈 [EMIM]Ac. Kinetic analysis showed that inoic liquid [EMIM]Et OSO3 and 2,2’- azino- bis（ 3-ethylbenzothiazoline-6- sulphonic acid）（ ABTS） shared the same active sites of the enzyme molecule. The inhibition was attributed to a possible competitive binding,suggesting the apparent activity of laccase in ILs was independent of the initial enzyme concentrations. Therefore,the increase of ABTS concentration was conducive for improving the utilizaiton efficiency of enzyme as well as the reaction rate.

关 键 词：漆酶 离子液体 侧耳菌糠 催化特性 抑制作用 

分 类 号：G242[文化科学]