Departamento de Química, Faculdade de Filosofia, Ciências E Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, São Paulo, 14040-901, Brazil.
Appl Biochem Biotechnol. 2022 Feb;194(2):848-861. doi: 10.1007/s12010-021-03688-5. Epub 2021 Sep 22.
Pectinases are widely used in a variety of industrial processes. However, their application is limited by low catalytic processivity, reduced stability, high cost, and poor re-use compatibility. These drawbacks may be overcome by enzyme immobilization with ferromagnetic nanoparticles, which are easily recovered by a magnetic field. In this work, an endopolygalacturonase from Chondrostereum purpureum (EndoPG) expressed in Pichia pastoris was immobilized on glutaraldehyde-activated chitosan ferromagnetic nanoparticles (EndoPG-MNP) and used to supplement a commercial enzyme cocktail. No significant differences in biochemical and kinetic properties were observed between EndoPG-MNP and EndoPG, although the EndoPG-MNP showed slightly increased thermostability. Cocktail supplementation with EndoPG-MNP increased reducing sugar release from orange wastes by 1.8-fold and showed a synergistic effect as compared to the free enzyme. Furthermore, EndoPG-MNP retained 65% of the initial activity after 7 cycles of re-use. These properties suggest that EndoPG-MNP may find applications in the processing of pectin-rich agroindustrial residues.
果胶酶广泛应用于各种工业过程中。然而，其应用受到低催化进程性、降低的稳定性、高成本和较差的重复使用兼容性的限制。通过用铁磁纳米粒子固定酶，可以克服这些缺点，铁磁纳米粒子可以很容易地通过磁场回收。在这项工作中，来自皱皮肿柄菇的内切多聚半乳糖醛酸酶（EndoPG）在毕赤酵母中表达，然后固定在戊二醛活化的壳聚糖铁磁纳米粒子（EndoPG-MNP）上，并用于补充商业酶混合物。尽管 EndoPG-MNP 显示出略微提高的热稳定性，但 EndoPG-MNP 和 EndoPG 的生化和动力学特性没有显著差异。与游离酶相比，用 EndoPG-MNP 补充酶混合物可使橙皮废物中的还原糖释放增加 1.8 倍，并显示出协同作用。此外，EndoPG-MNP 在重复使用 7 次后保留了初始活性的 65%。这些特性表明，EndoPG-MNP 可能在富含果胶的农业工业残渣的加工中有应用。
