基于无定形沸石-咪唑框架-8材料固定米根霉源脂肪酶研究
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(1.北京工商大学 食品与健康学院/北京食品营养与人类健康高精尖创新中心, 北京 100048;2.北京市食品添加剂工程技术研究中心, 北京 100048)

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Immobilization of Lipase Derived from Rhizopus oryzae Based on Amorphous Zeolitic Imidazolate Framework-8 Material
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(1.School of Food and Health/Beijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, Beijing 100048, China;2.Beijing Engineering and Technology Research Center of Food Additives, Beijing 100048, China)

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    摘要:

    多级孔沸石-咪唑框架-8(zeolitic imidazolate framework-8,ZIF-8)材料相较于传统的ZIF-8材料具有更大的孔径,在固定化酶领域展现了较好的应用前景。目前,利用多级孔ZIF-8材料固定米根霉源脂肪酶(Rhizopus oryzae lipase,ROL)的效果仍不清楚。通过调节ZIF-8材料前驱体的初始物质的量比例,制备得到一种无定形ZIF-8(amorphous zeolitic imidazolate framework-8,aZIF-8)材料,基于ZIF-8和aZIF-8材料, 采用原位自封装法制备得到固定化脂肪酶ROL@ZIF-8和ROL@aZIF-8。 利用红外光谱仪、X-射线衍射仪和扫描电子显微镜等研究了ZIF-8、aZIF-8、ROL@ZIF-8及ROL@aZIF-8的形貌及结构差异,比较了在不同ROL质量浓度条件下制备的ROL@aZIF-8与相同质量浓度ROL的酶活力,考察了ROL和ROL@aZIF-8的酸碱稳定性、温度耐受性、储藏稳定性以及对邻苯二甲酸二丁酯(dibutyl phthalate, DBP)和邻苯二甲酸二异丁酯(diisobutyl phthalate, DIBP)的水解能力。结果表明,aZIF-8材料呈现出包含十字花形、正方形、球形等多种形态的无定形形貌,是一种具有微孔和介孔的多级孔材料。 在ROL的质量浓度为4mg/mL的条件下制备的ROL@aZIF-8 呈现最大酶活力,为5.69U/mg。在相同的ROL质量浓度条件下,aZIF-8材料的疏水性及多级孔特性促使ROL@aZIF-8的酶活力比ROL高。ROL@aZIF-8比ROL具有更好的碱稳定性、温度耐受性、储藏稳定性以及更高的对DBP和DIBP的水解能力。

    Abstract:

    Hierarchical porous zeolitic imidazolate framework-8 (ZIF-8) materials have larger pore sizes compared with conventional ZIF-8 materials, and show better application prospects in the field of immobilized enzymes. At present, the effect of immobilizing Rhizopus oryzae lipase (ROL) using hierarchical porous ZIF-8 material remains unclear. In this study, an amorphous zeolitic imidazolate framework-8 (aZIF-8) material was prepared by adjusting the initial molar ratio of the precursor of ZIF-8 material, and the immobilized lipase ROL@ZIF-8 and ROL@aZIF-8 were prepared by in situ self-encapsulation based on the ZIF-8 and aZIF-8 materials, respectively. The morphological and structural differences of ZIF-8, aZIF-8, ROL@ZIF-8 and ROL@aZIF-8 were investigated by using infrared spectroscopy, X-ray diffractometer and scanning electron microscope, etc. The enzyme activities of ROL@aZIF-8 prepared under different ROL mass concentrations were compared with those of the same mass concentration of ROL. The acid-alkali stability, temperature tolerance, storage stability, and hydrolysis ability of ROL and ROL@aZIF-8 against dibutyl phthalate (DBP) and diisobutyl phthalate (DIBP) were investigated. The results showed that the aZIF-8 material presented an amorphous morphology containing various forms such as cruciate flower-like, square, and spherical, and was a hierarchical porous material with micropores and mesopores. ROL@aZIF-8 prepared at ROL mass concentration of 4mg/mL showed the maximum enzyme activity of 5.69U/mg. The hydrophobicity and hierarchical porous properties of the aZIF-8 material contributed to the higher enzyme activity of ROL@aZIF-8 than that of ROL at the same mass concentration of ROL. ROL@aZIF-8 showed better alkali stability, temperature tolerance, storage stability, and higher hydrolytic capacity for DBP and DIBP than ROL.

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芦瑜涵,张成楠,李秀婷,徐友强,李微微.基于无定形沸石-咪唑框架-8材料固定米根霉源脂肪酶研究[J].食品科学技术学报,2023,41(6):39-51.

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  • 收稿日期:2022-10-27
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  • 在线发布日期: 2023-11-07
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