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静电纺丝法固定化果糖基转移酶在可生物降解聚合物中的比较研究。

A Comparative Study on Immobilization of Fructosyltransferase in Biodegradable Polymers by Electrospinning.

机构信息

Institute of Technical Chemistry, Section of Carbohydrate Technology, Technical University Braunschweig, Gaußstraße 17, 38106, Braunschweig, Germany.

出版信息

Appl Biochem Biotechnol. 2018 Jul;185(3):847-862. doi: 10.1007/s12010-018-2694-6. Epub 2018 Jan 24.

Abstract

Commercial application of biocatalysts depends on the efficiency of the immobilization method and residual enzyme activity. Electrospinning offers a simple and versatile route to immobilize enzymes in submicron-sized fibers and thus improved mass transfer characteristics. Performance of encapsulation of fructosyltransferase from Bacillus subtilis by emulsion, suspension, and coaxial electrospinning was compared. We particularly focused on the effect of hydrophilic properties of a set of biodegradable polymers on support's activity. Bioactivity of electrospun support in aqueous medium increased in order of the matrix hydrophilicity. Additionally, the efficiency of electrospun fibers was compared with Sepabeads®, commercial epoxy-activated resins. In fibers, enzyme loading of 68.1 mg/g and specific enzyme activity of 5.5 U/mg was achieved compared to 49.5 mg/g and 2.2 U/mg on Sepabeads. Fructosyltransferase exhibited high sensitivity towards organic solvents and covalent attachment, respectively. Immobilization of native enzyme in coaxial fibers increased the specific activity to approx. 30 U/mg which corresponds to 24% of that of the free enzyme. Finally, operational stability of fiber supports was examined in a plug-flow reactor and 5% of initial substrate conversion remained after > 2000 cycles. The efficiency of core-shell immobilizates compared to one-dimensional fibers was both in batch and continuous reaction at least 4.4-fold higher.

摘要

商业应用生物催化剂取决于固定化方法的效率和残余酶活性。静电纺丝为在亚微米级纤维中固定化酶提供了一种简单而通用的方法,从而改善了传质特性。比较了通过乳液、悬浮液和同轴静电纺丝包封来自枯草芽孢杆菌的果糖基转移酶的性能。我们特别关注了一组可生物降解聚合物的亲水性对载体活性的影响。水介质中静电纺丝载体的生物活性按基质亲水性的顺序增加。此外,还比较了静电纺纤维与商业环氧活化树脂 Sepabeads®的效率。在纤维中,与 Sepabeads 上的 49.5mg/g 和 2.2U/mg 相比,酶载量达到了 68.1mg/g,比酶活达到了 5.5U/mg。果糖基转移酶对有机溶剂和共价结合分别具有高灵敏度。将天然酶固定在同轴纤维中,使比酶活提高到约 30U/mg,相当于游离酶的 24%。最后,在推流式反应器中考察了纤维载体的操作稳定性,在>2000 个循环后,仍有 5%的初始底物转化率。与一维纤维相比,核壳固定化剂在间歇和连续反应中的效率至少提高了 4.4 倍。

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