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磁性纳米粒子固定化皱褶假丝酵母脂肪酶在生物柴油酶化学联合酯化中的评价。

Evaluation of Candida rugosa Lipase Immobilized on Magnetic Nanoparticles in Enzymatic/Chemical Hydroesterification for Biodiesel Production.

机构信息

Department of Engineering of Bioprocesses and Biotechnology, School of Pharmaceutical Sciences, São Paulo State University (UNESP), Araraquara, SP, 14800-903, Brazil.

Department of Organic Chemistry, Center for Monitoring and Research of the Quality of Fuels, Biofuels, Crude Oil and Derivatives, Institute of Chemistry, São Paulo State University (UNESP), Araraquara, SP, 14800-900, Brazil.

出版信息

Appl Biochem Biotechnol. 2022 Nov;194(11):5419-5442. doi: 10.1007/s12010-022-04046-9. Epub 2022 Jul 5.

Abstract

This study aimed to (i) prepare functionalized maghemite nanoparticles for immobilization of Candida rugosa lipase (CRL) by covalent binding, (ii) evaluate the application of the immobilized derivative in the hydrolysis of waste cooking oil (WCO) to fatty acids, and (iii) assess the potential of the hydrolyzed material for biodiesel production by hydroesterification. Maghemite (γFeO) obtained by precipitation of FeCl with NHOH served as an efficient support for covalent immobilization of CRL. Fourier-transform infrared spectroscopy and hydrolytic activity analysis indicated that CRL was covalently immobilized on the surface of the maghemite support. The derivative showed an activity of 166.62 ± 8 U g in WCO hydrolysis at 40 °C and pH 6. Scanning electron microscopy revealed that, after lipase immobilization, nanoparticles became more dispersed, which is advantageous for biocatalysis reactions, as it increases the contact area with the substrate. WCO hydrolysis afforded 96 ± 0.2 wt% free fatty acids. In the second step, free fatty acids were subjected to chemical esterification with sulfuric acid, affording 94.4 ± 0.02 wt% fatty acid methyl esters (biodiesel). The findings of this study contribute to the field of biotechnology and may promote the development of enzymatic technologies for the synthesis of products of economic and social interest.

摘要

本研究旨在

(i)通过共价键合制备功能化的磁赤铁矿纳米粒子,用于固定化 Candida rugosa 脂肪酶(CRL);(ii)评估固定化衍生物在水解废食用油(WCO)制备脂肪酸中的应用;(iii)评估水解产物通过氢酯交换制备生物柴油的潜力。通过 NHOH 沉淀 FeCl 获得的磁赤铁矿(γFeO)作为 CRL 共价固定化的有效载体。傅里叶变换红外光谱和水解活性分析表明,CRL 已通过共价键合固定在磁赤铁矿载体表面。该衍生物在 40°C 和 pH 6 的条件下,对 WCO 水解的酶活为 166.62 ± 8 U g。扫描电子显微镜显示,脂肪酶固定化后,纳米颗粒变得更加分散,这有利于生物催化反应,因为它增加了与底物的接触面积。WCO 水解得到 96 ± 0.2 wt%的游离脂肪酸。在第二步中,游离脂肪酸与硫酸进行化学酯化,得到 94.4 ± 0.02 wt%的脂肪酸甲酯(生物柴油)。本研究的结果为生物技术领域做出了贡献,并可能促进具有经济和社会意义的产品的酶促技术的发展。

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