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贻贝仿生修饰多孔磁性材料的制备及其在固定化脂肪酶中的应用

李佥 王添誉 孙西同 陈晓艺 李苗 韩雨擎 曾祥冰 孙芳鸿 李宪臻

李佥, 王添誉, 孙西同, 等. 贻贝仿生修饰多孔磁性材料的制备及其在固定化脂肪酶中的应用[J]. 复合材料学报, 2024, 42(0): 1-14.
引用本文: 李佥, 王添誉, 孙西同, 等. 贻贝仿生修饰多孔磁性材料的制备及其在固定化脂肪酶中的应用[J]. 复合材料学报, 2024, 42(0): 1-14.
LI Qian, WANG Tianyu, SUN Xitong, et al. Preparation of mussel-inspired porous magnetic materials for application in immobilized lipase[J]. Acta Materiae Compositae Sinica.
Citation: LI Qian, WANG Tianyu, SUN Xitong, et al. Preparation of mussel-inspired porous magnetic materials for application in immobilized lipase[J]. Acta Materiae Compositae Sinica.

贻贝仿生修饰多孔磁性材料的制备及其在固定化脂肪酶中的应用

基金项目: 国家自然科学基金青年基金项目(31601411,21804129,22108024);辽宁省教育厅基本科研项目(JYTMS20230423)
详细信息
    通讯作者:

    李佥,博士,副教授,研究方向为贻贝仿生技术、生物纳米复合材料 E-mail:liqian19820903@163.com

    李宪臻,博士,教授,研究方向为微生物资源与生物催化 E-mail: bct-lab@dlpu.edu.cn

  • 中图分类号: Q814;TB332

Preparation of mussel-inspired porous magnetic materials for application in immobilized lipase

Funds: National Natural Science Foundation of China, China(31601411,21804129,22108024);Scientific Research Project of Liaoning Provincial Department of Education, China(JYTMS20230423)
  • 摘要: 本研究制备了一种以磁性壳聚糖为基材的多孔复合材料,通过在其表面涂覆聚多巴胺涂层替代传统的交联剂,用于脂肪酶的固定化研究。该材料具有优异的孔结构和较大的比表面积,孔容积可达0.6028 mL/g,比表面积可达106.8239 m2/g,经优化后固定化脂肪酶的酶活可达7392.91±121.22 U/g-载体。进一步探究了固定化酶的酶学性质,得到最佳反应温度为50 ℃,最佳反应pH为7.0,制备的固定化酶具有优异的热稳定性和pH稳定性,经过5次循环使用后,该固定化酶可以保持80%以上的初始酶活,经过10次循环使用后仍能保持52%的初始酶活。最后,将固定化酶应用于生物柴油转化并优化了相关工艺参数。

     

  • 图  1  Fe3O4(a)、Cs-Fe3O4(b)和聚多巴胺(PDA)-Cs-Fe3O4(c)的扫描电镜图及能谱图,Fe3O4(d)和PDA-Cs-Fe3O4(e)的透射电镜图

    Figure  1.  SEM and EDS images of Fe3O4 (a), Cs-Fe3O4 (b) and polydopamine (PDA)-Cs-Fe3O4 (c), TEM images of Fe3O4(d) and PDA-Cs-Fe3O4(e)

    图  2  Fe3O4、Cs-Fe3O4、PDA-Cs-Fe3O4和Lip-PDA-Cs-Fe3O4的红外光谱图

    Figure  2.  FTIR spectra of Fe3O4, Cs-Fe3O4, PDA-Cs-Fe3O4 and Lip-PDA-Cs-Fe3O4

    图  3  Lip-PDA-Cs-Fe3O4的X射线衍射谱图

    Figure  3.  X-ray spectrum of Lip-PDA-Cs-Fe3O4

    图  4  Fe3O4、Lip-PDA-Cs-Fe3O4的磁滞回线图谱

    Figure  4.  Hysteresis loop spectra of Fe3O4 and Lip-PDA-Cs-Fe3O4

    图  5  固定化温度对Lip-PDA-Cs-Fe3O4酶活的影响

    Figure  5.  Effect of immobilized temperature on the activity of Lip-PDA-Cs-Fe3O4

    图  6  固定化时间对Lip-PDA-Cs-Fe3O4酶活的影响

    Figure  6.  Effect of immobilized time on the activity of Lip-PDA-Cs-Fe3O4

    图  7  固定化pH对Lip-PDA-Cs-Fe3O4酶活的影响

    Figure  7.  Effect of immobilized pH on the activity of Lip-PDA-Cs-Fe3O4

    图  8  初始酶浓度对Lip-PDA-Cs-Fe3O4酶活的影响

    Figure  8.  Effect of initial enzyme concentration on the activity of Lip-PDA-Cs-Fe3O4

    图  9  游离酶与Lip-PDA-Cs-Fe3O4的最佳反应温度

    Figure  9.  Optimal reaction temperature of free enzyme and Lip-PDA-Cs-Fe3O4

    图  10  游离酶与Lip-PDA-Cs-Fe3O4的热稳定性

    Figure  10.  Thermal stability of free enzyme and Lip-PDA-Cs-Fe3O4

    图  11  游离酶与Lip-PDA-Cs-Fe3O4的最佳反应pH

    Figure  11.  Optimal reaction pH of free enzyme and Lip-PDA-Cs-Fe3O4

    图  12  游离酶与Lip-PDA-Cs-Fe3O4的pH稳定性

    Figure  12.  pH stability of free enzyme and Lip-PDA-Cs-Fe3O4

    图  13  Lip-PDA-Cs-Fe3O4的重复使用性

    Figure  13.  Reusability of Lip-PDA-Cs-Fe3O4

    图  14  Lip-PDA-Cs-Fe3O4的储藏稳定性

    Figure  14.  Storage stability of Lip-PDA-Cs-Fe3O4

    图  15  水油比对Lip-PDA-Cs-Fe3O4转化生物柴油的影响

    Figure  15.  Effect of water-oil ratio on the biodiesel conversion of Lip-PDA-Cs-Fe3O4

    图  16  加酶量对Lip-PDA-Cs-Fe3O4转化生物柴油的影响

    Figure  16.  Effect of enzyme quantity on the biodiesel conversion of Lip-PDA-Cs-Fe3O4

    图  17  反应时间对Lip-PDA-Cs-Fe3O4转化生物柴油的影响

    Figure  17.  Effect of reaction time on the biodiesel conversion of Lip-PDA-Cs-Fe3O4

    表  1  PDA-Cs-Fe3O4与Lip-PDA-Cs-Fe3O4的BET分析

    Table  1.   BET analysis of PDA-Cs-Fe3O4 and Lip-PDA-Cs-Fe3O4

    Name Surface area/(m2·g−1) Pore volume/(mL·g−1)
    PDA-Cs-Fe3O4 106.8239 0.6028
    Lip-PDA-Cs-Fe3O4 77.6027 0.4449
    下载: 导出CSV

    表  2  PDA-Cs-Fe3O4的孔径分析

    Table  2.   Pore size analysis of PDA-Cs-Fe3O4

    Pore size/nm Pore volume/(mL·g−1) Percentage/%
    Micropore 0.35-2 0.0081 1.35
    Mesopore 2-10 0.0589 9.84
    10-50 0.2175 36.31
    Macropore 50-120 0.3145 52.50
    下载: 导出CSV

    表  3  Lip-PDA-Cs-Fe3O4与其他研究中固定化酶热稳定性的比较

    Table  3.   Comparison of thermal stability of Lip-PDA-Cs-Fe3O4 with previous publications for immobilized lipase

    NameTemperature/℃Time/hRelative activity/%References
    PEG/PLA/CRL50℃2<70[39]
    L-PHM34<85[40]
    PFL@EMMS3<80[41]
    Lip-PDA-Cs-Fe3O4690This work
    Notes:Time is the duration of staying at 50℃; PEG is polyethylene glycol; PLA is polylactic acid; CRL is Candida rugosa lipase; L is Candida antarctica lipase; PHM is polyacrylamide hydrogel microspheres; 3 is the concentration of phosphate buffer solution of lipase immobilized on polyacrylamide hydrogel microspheres; PFL is Pseudomonas fluorescens lipase; EMMS is epoxy-functionalized macroporous and mesoporous SiO2.
    下载: 导出CSV

    表  4  Lip-PDA-Cs-Fe3O4与其他研究中固定化酶pH稳定性的比较

    Table  4.   Comparison of pH stability of Lip-PDA-Cs-Fe3O4 with previous publications for immobilized lipase

    NamepH rangeReferences
    CRL-BSA-CELLAs7.0-8.0[44]
    PPL@COF7.5-9.0[45]
    Fe3O4-COOH@UiO-66-NH2@PPL7.0-9.0[46]
    Lip-PDA-Cs-Fe3O46.0-9.0This work
    Notes:pH range is pH range in which the relative enzyme activity remains higher than 50%; CRL is Candida rugosa lipase; BSA is bovine albumin; CELLAs is cross-linked enzyme aggregates; PPL is porcine pancreatic lipase; COF is covalent organic framework; Fe3O4-COOH is carboxylic-functionalized magnetite; Uio-66-NH2 is zirconium aminobenzenedicarboxylate metal organic framework.
    下载: 导出CSV
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出版历程
  • 收稿日期:  2023-12-28
  • 修回日期:  2024-02-29
  • 录用日期:  2024-03-16
  • 网络出版日期:  2024-04-16

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