Cross-linked enzyme aggregate (CLEA) is a technology to overcome the limitation of enzymes for its application in chemical industries. The inability of repeated use of enzymes, stability and ease of separation from reaction mixture limits its applications. Here, magnetic combi-CLEA has been synthesised by adding amino-functionalized magnetic nanoparticles into pectinase ultra-clear (containing pectinases, xylanases and cellulases). Enzymes were precipitated on the surface of amino-functionalized magnetic nanoparticles with ethanol and cross-linked using glutaraldehyde. The structural characterization of magnetic combi-CLEA was studied by Scanning Electron Microscopy. Thermal stability was performed at 70 °C for pectinase and 80 °C for xylanase and cellulase respectively. Half-life (t1/2) of the xylanase, cellulase and pectinase in free form remarkably enhance from 84.51, 29.36, and 25.29 min respectively to 533.07, 187.29 and 147.44 min in magnetic-combi CLEA respectively. Magnetic combi-CLEA can be efficiently reused till 12th cycle after which pectinase, xylanase and cellulase retain 86.45%, 90.3% and 88.62% activity respectively. Using this CLEA preparation bioethanol concentration increases to 1.82-fold as compared to free enzyme, when simultaneous saccharification and fermentation was performed using wheat straw as the substrate. Magnetic combi-CLEA can be used for a variety of industrial applications like food processing, textile industry and bioethanol production.

中文翻译：

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可重复使用的纤维素酶和半纤维素酶磁性组合CLEA的制备与表征

交联酶聚合体（CLEA）是一种克服酶在化学工业中应用局限性的技术。酶不能重复使用、稳定性和易于从反应混合物中分离，限制了其应用。在这里，通过将氨基功能化的磁性纳米粒子添加到超透明果胶酶（含有果胶酶、木聚糖酶和纤维素酶）中合成了磁性组合 CLEA。酶用乙醇沉淀在氨基官能化磁性纳米粒子的表面，并用戊二醛交联。通过扫描电子显微镜研究了磁性组合-CLEA的结构特征。果胶酶的热稳定性分别在 70 °C 和 80 °C 下进行，木聚糖酶和纤维素酶。木聚糖酶的半衰期 (t1/2)，游离形式的纤维素酶和果胶酶在磁复合 CLEA 中分别从 84.51、29.36 和 25.29 分钟显着提高到 533.07、187.29 和 147.44 分钟。Magnetic combi-CLEA 可以有效地重复使用到第 12 个循环，之后果胶酶、木聚糖酶和纤维素酶分别保留 86.45%、90.3% 和 88.62% 的活性。当使用小麦秸秆作为底物同时进行糖化和发酵时，使用这种 CLEA 制剂生物乙醇浓度增加到游离酶的 1.82 倍。Magnetic combi-CLEA 可用于各种工业应用，如食品加工、纺织工业和生物乙醇生产。Magnetic combi-CLEA 可以有效地重复使用到第 12 个循环，之后果胶酶、木聚糖酶和纤维素酶分别保留 86.45%、90.3% 和 88.62% 的活性。当使用小麦秸秆作为底物同时进行糖化和发酵时，使用这种 CLEA 制剂生物乙醇浓度增加到游离酶的 1.82 倍。Magnetic combi-CLEA 可用于各种工业应用，如食品加工、纺织工业和生物乙醇生产。Magnetic combi-CLEA 可以有效地重复使用到第 12 个循环，之后果胶酶、木聚糖酶和纤维素酶分别保留 86.45%、90.3% 和 88.62% 的活性。当使用小麦秸秆作为底物同时进行糖化和发酵时，使用这种 CLEA 制剂生物乙醇浓度增加到游离酶的 1.82 倍。Magnetic combi-CLEA 可用于各种工业应用，如食品加工、纺织工业和生物乙醇生产。