Preparation of mussel-inspired porous magnetic materials for application in immobilized lipase
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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。Abstract: In this study, a porous composite material based on magnetic chitosan was prepared for lipase immobilization by applying a polydopamine coating on its surface instead of the traditional cross-linking agent. The material had excellent pore structure and large specific surface area, with a pore volume of0.6028 mL/g and a specific surface area of106.8239 m2/g, and the highest enzyme activity of the immobilized lipase could reach up to7392.91 ±121.22 U/g-carrier. In addition, the enzymatic properties of the immobilized enzyme were investigated, and the optimal reaction temperature and pH were 50 ℃ and 7.0, respectively. Furthermore, the prepared immobilized enzyme exhibited good thermal and pH stabilities, and could maintain above 80% of the initial enzyme activity by the immobilized enzyme after five cycles of reuse, while 52% of the initial enzyme activity could be still maintained after ten cycles of reuse. Finally, the immobilized enzyme was applied in biodiesel conversion and the process was optimized.-
Key words:
- Magnetic materials /
- Mussel-inspired /
- Lipase /
- Enzymatic properties /
- Biodiesel conversion
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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 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 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
Name Temperature/℃ Time/h Relative activity/% References PEG/PLA/CRL 50℃ 2 <70 [39] L-PHM3 4 <85 [40] PFL@EMMS 3 <80 [41] Lip-PDA-Cs-Fe3O4 6 90 This 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. 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
Name pH range References CRL-BSA-CELLAs 7.0-8.0 [44] PPL@COF 7.5-9.0 [45] Fe3O4-COOH@UiO-66-NH2@PPL 7.0-9.0 [46] Lip-PDA-Cs-Fe3O4 6.0-9.0 This 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. -
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