MOFs固定5-羟甲基糠醛氧化酶及其催化活性的研究 <sup>*</sup>

doi:10.13523/j.cb.20190606

中国生物工程杂志

2019, Vol. 39

Issue (6): 41-47 DOI: 10.13523/j.cb.20190606

研究报告

MOFs固定5-羟甲基糠醛氧化酶及其催化活性的研究 ^*

巩凤芹^1,²,刘启顺²,谭海东²,金花³,谭成玉^1,^**(

),尹恒^2,^**(

)

1 大连海洋大学海洋科技与环境学院大连 116023
2 中国科学院大连化学物理研究所天然产物与糖工程组大连 116023
3 宁波大学材料科学与化学工程学院宁波 315211

Immobilization of 5-Hydroxymethylfurfural Oxidase within MOFs for Catalysis

Feng-qin GONG^1,²,Qi-shun LIU²,Hai-dong TAN²,hua JIN³,Cheng-yu TAN^1,^**(

),Heng YIN^2,^**(

)

1 College of Marine Science and Environment, Dalian Ocean University, Dalian 116023, China
2 Dalian Institute of Chemical Physics, Chinese Academy of Sciences, CAS, Dalian 116023, China
3 College of Material and Engineering, Ningbo University, Ningbo 315211, China

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摘要：

固定化酶作为一种绿色高效的生物催化剂,其性能远超游离酶。目前酶的固定化技术适用范围仍然较小,酶的研究范围多停留在模型酶阶段,扩大固定化酶的研究范围具有十分重要的意义。金属有机骨架材料(MOFs)作为酶固定化的载体在近些年得到了广泛的探索,但是具有生物功能的酶-MOFs复合材料的许多特性仍有待挖掘。采用仿生矿化的合成方法将5-羟甲基糠醛氧化酶(HMFO)固定到以沸石咪唑酯(ZIF-8)为代表的MOFs材料中,制备得到一种新的生物催化剂HMFO@ZIF-8,扫描电子显微镜表征其形态区别于经典的菱形十二面体。采用考马斯亮蓝法测定蛋白质浓度,计算得到酶的固定化效率达到89.0%。HMFO@ZIF-8催化5-羟甲基糠醛的转化率达到84.3%,收率和选择性均高于游离酶。拓展了MOFs固定化酶的研究范围,为研究其他生物大分子复合材料的生物催化剂提供一定的借鉴意义。

关键词： 金属有机骨架材料; 5-羟甲基糠醛氧化酶; 固定化酶; 生物催化剂

Abstract:

Immobilized enzymes are biocatalysts that typically have higher functional efficiency and reproducibility than free enzymes. However, the use of enzyme immobilization is still limited, and the research is mainly conducted within model enzymes. In recent years, the study of metal organic frameworks (MOFs) as enzyme immobilization carriers has attracted great attention, and the nature of MOF-enzyme complexes remains to be elucidated. In the present study, a novel immobilized biocatalyst was prepared by embedding a 5-hydroxymethylfurfural oxidase (HMFO) in zeolite imidazole ester skeleton ZIF-8(a typical MOF) through biomimetic mineralization. The morphology of the composite catalyst under scanning electron microscope was different from the classical rhombohedral dodecahedron of MOF immobilized enzyme. Protein concentration assay conducted by Commassie brilliant blue G-250 method indicated that the immobilization efficiency was reached 89.0%. Moreover, the conversion efficiency of HMFO@ZIF-8 to 5-hydroxymethylfurfural reached 84.3%, and the yield and selectivity were significantly improved as compared to free HMFO.New knowledge for the research of MOFs immobilized enzymes and facilitates the development of immobilized biomacromolecules and synthetic biocatalysts were provided.

Key words: MOFs 5-Hydroxymethylfurfural oxidase Immobilized enzyme Biocatalyst

收稿日期: 2018-10-30 出版日期: 2019-07-12

ZTFLH:

Q814

基金资助: * 国家自然科学基金(31670803);大连化物所创新基金资助项目(DICP ZZBS201704);大连化物所创新基金资助项目(DICP BioChE-X201801)

通讯作者: 谭成玉,尹恒 E-mail: tanchyu@dlou.edu.cn;yinheng@dicp.ac.cn

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引用本文:

巩凤芹,刘启顺,谭海东,金花,谭成玉,尹恒. MOFs固定5-羟甲基糠醛氧化酶及其催化活性的研究 ^*[J]. 中国生物工程杂志, 2019, 39(6): 41-47.

Feng-qin GONG,Qi-shun LIU,Hai-dong TAN,hua JIN,Cheng-yu TAN,Heng YIN. Immobilization of 5-Hydroxymethylfurfural Oxidase within MOFs for Catalysis. China Biotechnology, 2019, 39(6): 41-47.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20190606 或 https://manu60.magtech.com.cn/biotech/CN/Y2019/V39/I6/41

图1 HMFO@ZIF-8的负载量

图2 HMFO和HMFO@ZIF-8的SDS-PAGE分析图

图3 FITC-HMFO@ZIF-8激光共聚焦显微镜图

图4 HMFO@ZIF-8(a)、BSA@ZIF-8(b)的扫描电镜图

图5 不同浓度的2-甲基咪唑对HMFO酶活的影响

图6 不同浓度的乙酸锌对HMFO酶活的影响

图7 HMFO和HMFO@ZIF-8催化HMF后的色谱图

图8 HMF、DFF、FFA的标准曲线

表1 HMFO@ZIF-8催化HMF的转化率

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