一种通过固定在铜铁氧体纳米颗粒上来提高漆酶活性以降解木质素的新方法。
A novel method for improving laccase activity by immobilization onto copper ferrite nanoparticles for lignin degradation.
机构信息
Bioenergy Research Laboratory, Department of Biotechnology, Bannari Amman Institute of Technology, Sathyamangalam, Erode, Tamil Nadu, India.
Bioenergy Research Laboratory, Department of Biotechnology, Bannari Amman Institute of Technology, Sathyamangalam, Erode, Tamil Nadu, India.
出版信息
Int J Biol Macromol. 2020 Jun 1;152:1098-1107. doi: 10.1016/j.ijbiomac.2019.10.198. Epub 2019 Nov 18.
In this paper, a novel method for simultaneous enhancement of catalytic activity and reusability of laccase was carried out to overcome the limitations on industrial application of laccase. The immobilization of laccase onto copper ferrite magnetic nanoparticles (CuMNPs) and ferrite magnetic nanoparticles (MNPs) were optimized at 50 mM glutaraldehyde concentration and 1:5 enzyme:nanoparticles (NPs) ratio for 9 h of cross-linking time, yielding a maximum activity recovery of 94.68 ± 0.92% and 89.78 ± 1.24%, respectively. The laccase immobilized NPs were characterized using physico-chemical methods such as SEM-EDAX, FTIR, XRD, TGA and VSM and the laccase immobilized CuMNPs showed 18% higher activity as compared to free enzyme. The prepared CuMNPs and MNPs showed superior thermal stability (50-70 °C) with t increased by 5.7 and 4.1 folds, respectively, as compared to free laccase. The laccase in immobilized forms exhibited higher kinetic potential and stable at wide temperature and pH range. In addition, laccase immobilized NPs retained more than 70% residual activity during reuse up to 6 cycles and storing for 20 days at 4 °C. The laccase immobilized CuMNPs showed higher delignification (43.28 ± 1.46%) on Ipomoea carnea than laccase immobilized MNPs.
本文提出了一种同时提高漆酶催化活性和可重复使用性的新方法，以克服漆酶在工业应用中的局限性。通过优化戊二醛浓度为 50mM 和酶与纳米粒子（NPs）比例为 1:5，交联时间为 9 小时，实现了漆酶固定化到铜铁氧体磁性纳米粒子（CuMNPs）和铁氧体磁性纳米粒子（MNPs）上，最大酶活回收率分别为 94.68±0.92%和 89.78±1.24%。采用 SEM-EDAX、FTIR、XRD、TGA 和 VSM 等物理化学方法对固定化 NPs 进行了表征，固定化 CuMNPs 的酶活比游离酶高 18%。与游离酶相比，制备的 CuMNPs 和 MNPs 具有更好的热稳定性（50-70°C），t 值分别增加了 5.7 和 4.1 倍。固定化酶在较宽的温度和 pH 范围内具有更高的动力学潜力和稳定性。此外，固定化 NPs 的酶在重复使用 6 次和在 4°C 下储存 20 天的过程中，保留了超过 70%的残余活性。固定化 CuMNPs 的漆酶在Ipomoea carnea 上的木质素脱除率（43.28±1.46%）高于固定化 MNPs 的漆酶。
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