广西师范大学学报（自然科学版） ›› 2022, Vol. 40 ›› Issue (2): 158-169.doi: 10.16088/j.issn.1001-6600.2021032201

• • 上一篇    下一篇

裂解多糖单加氧酶纳米花固定化研究

任文文, 孙云泽, 李蓉*   

  1. 大连工业大学 生物工程学院,辽宁 大连 116000
  • 收稿日期:2021-03-22 修回日期:2021-04-30 发布日期:2022-05-31
  • 通讯作者: 李蓉(1987—),女,浙江奉化人,大连工业大学讲师,博士。E-mail: ryong_li@126.com
  • 基金资助:
    国家自然科学基金(31600640);辽宁省教育厅科学研究项目(J2019015)

Study on Lytic Polysaccharide Monooxygenase Nanoflower for Immobilization

REN Wenwen, SUN Yunze, LI Rong*   

  1. School of Biological Engineering,Dalian Polytechnic University,Dalian Liaoning 116000, China
  • Received:2021-03-22 Revised:2021-04-30 Published:2022-05-31

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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=7.4 0.01 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。

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

Abstract: 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.

Key words: immobilization, nanoflower, lysing polysaccharide monooxygenase, enzymatic properties, inorganic crystal complex

中图分类号: 

  • Q814.2
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