Resilient Enzymes through Immobilisation: Stable NDP Polyphosphate Phosphotransferase from Ruegeria pomeroyi for Nucleotide Regeneration

IF 4.7 3区材料科学 Q1 ENGINEERING, ELECTRICAL & ELECTRONIC ACS Applied Electronic Materials Pub Date : 2024-02-24 DOI:10.3390/catal14030165

Kristin Hölting, Sebastian Götz, M. Aßmann, P. Bubenheim, Andreas Liese, Jürgen Kuballa

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Abstract

Immobilisation plays an important role in the industrial application of enzymes. The stabilisation and reusability of immobilised enzymes reduce the cost of the catalyst and facilitate their use in continuously operated reactors. For this purpose, an applicable type of immobilisation needs to be identified. In this study, we investigate the conversion of CDP and PolyP to CTP by NDP polyphosphate phosphotransferase 3 from Ruegeria pomeroyi (RpPPK2-3) and describe the covalent immobilisation of RpPPK2-3. In order to select a suitable carrier for the immobilisation of RpPPK2-3, a screening with different amino methacrylate (glutaraldehyde-pre-activated) and epoxy methacrylate carriers was carried out. The epoxy methacrylate carrier ECR8209M (Purolite®) was found to be the most suitable. With a half-life of 462 d when stored at 6 °C and a 50-fold reusability with a 93% residual activity, the immobilised enzyme showed a higher stability compared to the soluble enzyme with a half-life of 0.04 d. Although the half-life of the soluble enzyme could be increased to 32 d by adding PPi, it could not reach the stability of the immobilisate. Due to the resilience of the immobilisate, it is suitable for application in continuous reactor set-ups, e.g., packed-bed reactors.

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通过固定化实现弹性酶：用于核苷酸再生的 Ruegeria pomeroyi 稳定 NDP 多磷酸磷酸转移酶

固定化在酶的工业应用中发挥着重要作用。固定化酶的稳定性和可重复使用性降低了催化剂的成本，便于在连续运行的反应器中使用。为此，需要确定一种适用的固定化类型。在本研究中，我们研究了 Ruegeria pomeroyi（RpPPK2-3）的 NDP 多磷酸磷酸转移酶 3 将 CDP 和 PolyP 转化为 CTP 的过程，并描述了 RpPPK2-3 的共价固定化。为了选择适合固定 RpPPK2-3 的载体，研究人员筛选了不同的甲基丙烯酸氨基酯（戊二醛预活化）和甲基丙烯酸环氧酯载体。结果发现环氧甲基丙烯酸酯载体 ECR8209M（Purolite®）最为合适。与半衰期为 0.04 d 的可溶性酶相比，固定化酶显示出更高的稳定性。虽然通过添加 PPi，可溶性酶的半衰期可延长至 32 d，但仍无法达到固定化酶的稳定性。由于固定化物的弹性，它适合应用于连续反应器装置，如填料床反应器。

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来源期刊

ACS Applied Electronic Materials Multiple-

CiteScore

7.20

自引率

4.30%

发文量

567

期刊介绍： ACS Applied Electronic Materials is an interdisciplinary journal publishing original research covering all aspects of electronic materials. The journal is devoted to reports of new and original experimental and theoretical research of an applied nature that integrate knowledge in the areas of materials science, engineering, optics, physics, and chemistry into important applications of electronic materials. Sample research topics that span the journal's scope are inorganic, organic, ionic and polymeric materials with properties that include conducting, semiconducting, superconducting, insulating, dielectric, magnetic, optoelectronic, piezoelectric, ferroelectric and thermoelectric. Indexed/Abstracted： Web of Science SCIE Scopus CAS INSPEC Portico

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