β‐glucosidase (BglA) immobilization from Thermotoga maritima on magnetic nanoparticles (MNPs) functionalized with chitosan (Cs) were efficiently investigated to improve lactose conversion and galactooligosaccharides (GOS) production. We used a batch method in order to improve the conversion of lactose to GOS. The efficiency and yield of immobilization were 79% and immobilized BglA was effectively recycled via a magnetic separation procedure through a batch‐wise GOS with no activity lessening. Furthermore, analyses were done through screening kinetics of enzyme activity, sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE), Fourier transform infrared spectroscopy (FT‐IR), and transmission electron microscopy (TEM). Proposed methodology of immobilization shows a potential application as it is stable which was proved through many methods including pH, temperature, heat treatment, storage, and kinetics of the enzyme. GOS and residual enzyme activity showed to be 28.76 and 40.44%, respectively. However, free enzyme synthesis of GOS yield was just 24% after 12 hr. This study proposed applying magnet in the immobilization process of BglA on Cs‐MNPs to produce GOS as new method for immobilizing enzyme in a biostable and cost‐efficient way.

中文翻译：

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使用固定在壳聚糖功能化的磁性纳米粒子上的β-葡萄糖苷酶大批量生产低聚半乳糖

滨海嗜热菌的β-葡萄糖苷酶（BglA）固定化有效地研究了壳聚糖（Cs）功能化的磁性纳米颗粒（MNP）的性能，以改善乳糖转化和低聚半乳糖（GOS）的生产。为了提高乳糖向GOS的转化率，我们使用了分批方法。固定化的效率和产率为79％，固定化的BglA通过磁分离程序通过间歇式GOS有效回收，且活性未降低。此外，还通过筛选酶活性动力学，十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE），傅立叶变换红外光谱（FT-IR）和透射电子显微镜（TEM）进行了分析。拟议的固定化方法具有稳定的潜在应用前景，并已通过许多方法证明了这一应用，包括pH值，温度，热处理，储存，和酶的动力学。GOS和残留酶活性分别为28.76％和40.44％。但是，在12小时后，游离酶合成的GOS产量仅为24％。这项研究提出在BglA固定化Cs-MNPs的过程中使用磁铁生产GOS，以此作为一种以生物稳定且经济高效的方式固定酶的新方法。