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将嗜热脂环酸芽孢杆菌的葡萄糖异构酶固定在 Sepabeads EC-HA 上及其在填充床反应器中连续生产高果糖糖浆中的有效应用。

Immobilization of the glucose isomerase from Caldicoprobacter algeriensis on Sepabeads EC-HA and its efficient application in continuous High Fructose Syrup production using packed bed reactor.

机构信息

Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia.

Institute of Catalysis and Petrochemistry, CSIC, Marie Curie 2, 28049 Madrid, Spain.

出版信息

Food Chem. 2020 Mar 30;309:125710. doi: 10.1016/j.foodchem.2019.125710. Epub 2019 Oct 19.

Abstract

The glucose isomerase GICA from Caldicoprobacter algeriensis was immobilized by ionic adsorption on polymethacrylate carriers (Sepabeads EC-EA and EC-HA) or covalent attachment to glyoxal agarose. The Sepabeads EC-HA yielded the highest recovery of activity (89%). The optimum temperature and pH of immobilized GICA were 90 °C and 7.0, respectively, similar to the corresponding values of free enzyme. Nevertheless, the adsorbed enzyme displayed higher relative activity at acidic pH, greater thermostability, and better storage stability, compared to the free form. Moreover, the immobilized enzyme showed an excellent operational stability, in 15 successive 3 h reaction cycles at 85 °C under a batch reactor, preserving 83% of its initial activity. Interestingly, a continuous process for High Fructose Syrup (HFS) production was established with the adsorbed GICA using a packed bed reactor during eleven days at 70 °C. HPAEC-PAD analysis showed a maximum bioconversion rate of 49% after 48 h of operation.

摘要

从 Algeriensis 热脂环芽孢杆菌中分离得到的葡萄糖异构酶 GICA 可以通过离子吸附固定在聚甲基丙烯酸酯载体(Sepabeads EC-EA 和 EC-HA)上,或者通过共价键连接到乙二醛琼脂糖上。Sepabeads EC-HA 产生了最高的酶活回收率(89%)。固定化 GICA 的最适温度和 pH 值分别为 90°C 和 7.0,与游离酶的相应值相似。然而,与游离酶相比,吸附酶在酸性 pH 值下显示出更高的相对活性、更好的热稳定性和更好的储存稳定性。此外,固定化酶在分批反应器中 85°C 下连续 15 个 3 小时反应循环中表现出极好的操作稳定性,保持了初始活性的 83%。有趣的是,使用填充床反应器在 70°C 下进行了 11 天的实验,建立了使用吸附 GICA 生产高果糖糖浆(HFS)的连续工艺。HPAEC-PAD 分析显示,在 48 小时的运行后,生物转化率达到了最大的 49%。

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