裂解多糖单加氧酶纳米花固定化研究  

Study on Lytic Polysaccharide Monooxygenase Nanoflower for Immobilization

作　　者：任文文 孙云泽 李蓉 REN Wenwen;SUN Yunze;LI Rong(School of Biological Engineering,Dalian Polytechnic University,Dalian Liaoning 116000,China)

机构地区：[1]大连工业大学生物工程学院,辽宁大连116000

出　　处：《广西师范大学学报（自然科学版）》2022年第2期158-169,共12页Journal of Guangxi Normal University:Natural Science Edition

基　　金：国家自然科学基金(31600640);辽宁省教育厅科学研究项目(J2019015)。

摘　　要：本文利用新型无机晶体复合物磷酸铜作为载体,对裂解多糖单加氧酶cx-LPMO-B进行固定化,研究其固定化过程的最适条件,并对游离酶及固定化酶的酶学性质进行对比。结果显示:在搅拌条件下,向含有裂解多糖单加氧酶pH=7.40.01 mol/L的磷酸盐缓冲溶液中滴加硫酸铜,可形成酶-磷酸铜纳米花,即cx-LPMO-B-NF;在25℃、14 h、酶含量为0.3 g/L条件下制备得到的cx-LPMO-B-NF活性最高,固定化酶重复使用6次后仍能保持60%以上的酶活;扫描电子显微镜(SEM)及透射电子显微镜(TEM)发现cx-LPMO-B-NF结构呈现分散均匀且单一的盛开花朵状;固定化酶最适反应pH=4.0,最适反应温度50℃。固定化酶重复使用性显著增强,花状结构增加了其表面积,更加有利于对游离酶的固定化,反应条件较温和使其具有较高的工业化应用前景。In this paper,a novel inorganic crystal complex copper phosphate was used as a carrier to immobilize the lytic polysaccharide monooxygenase(cx-LPMO-B),the optimal conditions for immobilization of cx-LPMO-B was investigated,and the enzymatic properties were compared with the free and immobilization of cx-LPMO-B.The results showed that,copper sulfate was added to the pH=7.4 and 0.01 mol/L phosphate buffer solution containing the cx-LPMO-B to form immobilized cx-LPMO-B-copper phosphate nanoflowers,namely cx-LPMO-B-NF.The cx-LPMO-B-NF reached the highest immobilized yield at 25℃for 14 h with the 0.3 g/L cx-LPMO-B.The immobilized cx-LPMO-B-NF retained a residual activity of approximately 60%after 6 cycles reaction.The nano structure of the immobilized cx-LPMO-B-NF was characterized with scanning electron microscope(SEM)and transmission electron microscope(TEM).It was performed that the immobilized cx-LPMO-B-NF was uniformly dispersed and single blooming flower shape.The optimal pH and temperature for immobilized cx-LPMO-B-NF were pH=4.0 and 50℃,respectively.The nano flower structure increased surface area of immobilized carrier to enhance cx-LPMO-B immobilization.Moreover,the reusability of immobilized cx-LPMO-B-NF was significantly enhanced.Therefore,the immobilized cx-LPMO-B-NF can promote advantages for cx-LPMO-B industrial application.

关 键 词：固定化 纳米花 裂解多糖单加氧酶 酶学性质 无机晶体复合物 

分 类 号：Q814.2[生物学—生物工程]