Immobilization of bromelain on cobalt-iron magnetic nanoparticles (CoFe2O4) for casein hydrolysis

IF 0.2 Q4 CHEMISTRY, MULTIDISCIPLINARY

Revista Colombiana de Quimica Pub Date : 2020-01-01 DOI:10.15446/rev.colomb.quim.v1n49.69474

José Mauricio García Colmenares, Julia Constanza Reyes Cuellar

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引用次数: 3

Abstract

By means of recycling an enzyme, bromelain was used in casein hydrolysis facilitated by a nanobiocatalyst consisting of bromelain, CoFe2O4 magnetic nanoparticles, chitosan, and glutaraldehyde. Bromelain was immobilized on the chitosan cobalt-magnetite nanoparticle surface via covalent bonds to form the nanobiocatalyst. Immobilized bromelain showed 77% immobilization binding, retaining 85 ± 2% of the initial catalytic activity. Nanoparticles and immobilized bromelain were characterized using UV-Vis and IR spectroscopies, X-ray, square wave voltammetry (SWV), cyclic voltammetry (CV), vibrating-sample magnetization (VSM), and transmission electron microscope (TEM). The Michaelis-Menten constant (KM) and VMAX of the free and immobilized enzyme were calculated: KM = 2.1 ± 0.18 mM and 1.8 mM, respectively and VMAX = 6.08 x 10-2 ± 2.1 x 10-2 U/min and 6.46 ± 0.91 U/min, respectively. The thermal stability of the free enzyme was higher than the immobilized enzyme: 95-98% and 83-87%, respectively. An optimum pH of 6 and a temperature of 20 °C were determined in both cases. Immobilized bromelain maintained 50% of the initial catalytic activity after the fifth use. The immobilized bromelain proved to be effective and reusable for casein hydrolysis. As novel contribution the characterization by VOC and CV was carried out.

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用钴铁磁性纳米粒子（CoFe2O4）固定化菠萝蛋白酶水解酪蛋白

通过酶的循环利用，以菠萝蛋白酶、CoFe2O4磁性纳米颗粒、壳聚糖和戊二醛组成的纳米生物催化剂促进酪蛋白的水解。将菠萝蛋白酶通过共价键固定在壳聚糖钴磁铁矿纳米颗粒表面，形成纳米生物催化剂。固定化菠萝蛋白酶的固定化结合率为77%，保留了85±2%的初始催化活性。采用紫外可见光谱(UV-Vis)和红外光谱(IR)、x射线、方波伏安法(SWV)、循环伏安法(CV)、振动样品磁化法(VSM)和透射电镜(TEM)对纳米颗粒和固定化菠萝蛋白酶进行了表征。计算游离酶和固定化酶的Michaelis-Menten常数(KM)和VMAX: KM分别为2.1±0.18 mM和1.8 mM, VMAX分别为6.08 × 10-2±2.1 × 10-2 U/min和6.46±0.91 U/min。游离酶的热稳定性高于固定化酶，分别为95-98%和83-87%。在这两种情况下，确定了最佳pH为6，温度为20°C。固定化菠萝蛋白酶在第五次使用后仍保持50%的初始催化活性。结果表明，固定化菠萝蛋白酶对酪蛋白的水解是有效且可重复使用的。作为新的贡献，用VOC和CV进行了表征。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Revista Colombiana de Quimica CHEMISTRY, MULTIDISCIPLINARY-

CiteScore

1.50

自引率

0.00%

发文量

期刊介绍： The Revista Colombiana de Química - Colombian Journal of Chemistry (Rev. Colomb. Quim.) Is a peer-reviewed scientific journal of the Department of Chemistry, Faculty of Sciences of the Universidad Nacional de Colombia, Bogotá. It currently publishes three volumes per year: January-April, May-August and September-December. All the content published by the Journal (available online) is under a Creative Commons attribution license type BY 4.0, that allows any person or entity in the world to freely access the content, share it, download it, adapt it or make derivative works without any restriction, provided that it adequately indicates the credit to the authors and to the Colombian Journal of Chemistry.