Immobilized protease on the magnetic nanoparticles used for the hydrolysis of rapeseed meals
Immobilized protease on the magnetic nanoparticles used for the hydrolysis of rapeseed meals
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DOI：
10.1016/j.jmmm.2010.01.029
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发表时间：
2010-07-01
影响因子：
2.7
通讯作者：
Chen, Hong
中科院分区：
文献类型：
--
作者：
Jin, Xin;Li, Ju-Fang;Chen, Hong
采用水相共沉淀法合成了平均粒径为25.4 nm的(3-氨丙基)三乙氧基硅烷及其改性磁性纳米粒子。然后以1，4-苯二异氰酸酯为偶联剂，将这些纳米粒子与碱性蛋白酶进行共价偶联。实验表明，固定化蛋白水解酶具有较高的催化活性，以酪蛋白为底物时，其催化活性可达47.8%。结果表明，固定化酶在磁性纳米粒子上的催化活性在60d后仍能保持98.63+/-2.37%。在货架期试验中，它比游离的蛋白酶更稳定。最适反应温度、最适pH等酶反应条件与游离蛋白酶相同。混合分离实验表明，经过生物催化处理后的固定化酶可以通过磁性纳米颗粒回收利用。以菜籽粕为底物，固定化碱性蛋白酶水解度为9.86%，游离水解度为10.41%。利用固定化蛋白酶将菜籽粕中的大分子蛋白质水解成多肽或氨基酸等小分子物质。因此，基于这些磁性纳米粒子，为酶的回收利用在活性多肽的连续生产中的应用提供了一条新的、高效、经济的途径。(C)2010爱思唯尔B.V.保留所有权利。
(3-aminopropl) triethoxysilaneand modified magnetic nanoparticles with the average diameter of 25.4 nm were synthesized in water-phase co-precipitation method. And then these nanoparticles were covalently coupled with alkaline protease as enzyme carrier by using 1,4-phenylene diisothlocyanate as coupling agent. Experiments showed that the immobilized protease can keep the catalytic bioactivity, which can reach to 47.8% when casein was served as substrate. Results showed that the catalytic activity of immobilized protease on these magnetic nanoparticles could retain 98.63 +/- 2.37% after 60 days. And it is more stable than the free protease during the shelf-life test. The enzyme reaction conditions such as optimum reaction temperature and pH are the same as free protease. Furthermore, mix-and-separate experiments showed that the immobilized protease could be recycled through the magnetic nanoparticles after the biocatalysis process. When the rapeseed meals were used as substrate, the degree of hydrolysis of immobilized alkaline protease achieved 9.86%, while it was 10.41% for the free protease. The macromolecular proteins of rapeseed meals were hydrolyzed by immobilized protease into small molecules such as polypeptides or amino acids. Thus, a novel efficient and economic way for the recycling of enzymes in the application of continuous production of active peptides was provided based on these magnetic nanoparticles. (C) 2010 Elsevier B.V. All rights reserved.