Enzyme immobilization has been known to be one of the methods to improve the stability and reusability of enzyme. In this study, a strategy to optimize laccase immobilization on polyethylene terephthalate grafted with maleic anhydride electrospun nanofiber mat (PET-g-MAH ENM) was developed. The development involves the screening and optimization processes of the crucial factors that influence the immobilization yield such as enzyme concentration, pH values, covalent bonding (CV) time, CV temperature, crosslinking (CL) time, CL temperature and glutaraldehyde concentration using two-level factorial design and Box-Behnken design (BBD), respectively. It was found that laccase concentration, pH values and glutaraldehyde concentration play important role in enhancing the immobilization yield of laccase on PET-g-MAH ENM in the screening process. Subsequently, the optimization result showed at 0.28 mg/ml laccase concentration, pH 3 and 0.45% (v/v) glutaraldehyde concentrations gave the highest immobilization yield at 87.64% which was 81.2% increment from the immobilization yield before optimization. Under the optimum condition, the immobilized laccase was able to oxidize 2, 2-azino-bis 3-ethylbenzothiazoline-6- sulfonic acid (ABTS) in a broad range of pH (pH 3-6) and temperature (20- 70 °C). Meanwhile, the kinetic parameters for Km and Vmax were 1.331 mM and 0.041 mM/min, respectively. It was concluded that the optimization of immobilized laccase on PET-g-MAH ENM enhance the performance of this biocatalyst.

译文

酶固定化已被认为是提高酶的稳定性和可重复使用性的方法之一。在这项研究中,开发了一种优化在马来酸酐电纺纳米纤维毡 (PET-g-MAH ENM) 接枝的聚对苯二甲酸乙二醇酯上固定漆酶的策略。该开发涉及影响固定化产率的关键因素的筛选和优化过程,例如酶浓度,ph值,共价键合 (CV) 时间,CV温度,交联 (CL) 时间,CL温度和戊二醛浓度分别采用二级析因设计和Box-Behnken设计 (BBD)。研究发现,在筛选过程中,漆酶浓度,ph值和戊二醛浓度对提高漆酶在PET-g-MAH ENM上的固定化率具有重要作用。随后,优化结果显示在0.28 mg/ml漆酶浓度下,pH 3和0.45% (v/v) 戊二醛浓度在87.64% 时给出了最高的固定化产率,比优化前的固定化产率增加了81.2%。在最佳条件下,固定化的漆酶能够在较宽的pH (pH 3-6) 和温度 (20- 70 °C) 范围内氧化2,2-氮杂-双3-乙基苯并噻唑啉-6-磺酸 (ABTS)。同时,Km和Vmax的动力学参数分别为1.331 mM和0.041 mM/min。结论是,在PET-g-MAH ENM上固定化漆酶的优化可增强该生物催化剂的性能。

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