通过位点特异性聚乙二醇化改善重组嗜热栖热放线菌聚半乳糖醛酸酶的性质。

Improving Properties of Recombinant SsoPox by Site-Specific Pegylation.

作者信息

Parikh Harsh, Bajaj Priyanka, Tripathy Rajan K, Pande Abhay H

机构信息

Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), Sector 67, S.A.S. Nagar, (Mohali)-160 062, Punjab, India.

出版信息

Protein Pept Lett. 2015;22(12):1098-103. doi: 10.2174/0929866522666151002122751.

DOI:10.2174/0929866522666151002122751

PMID:26428299

Abstract

SsoPox, a ~35 kDa enzyme from Sulfolobus solfataricus, can hydrolyze and inactivate a variety of organophosphate (OP)-compounds. The enzyme is a potential candidate for the development of prophylactic and therapeutic agent against OP-poisoning in humans. However, the therapeutic use of recombinant SsoPox suffers from certain limitations associated with the use of recombinant protein pharmaceuticals. Some of these limitations could be overcome by conjugating SsoPox enzyme with polyethylene glycol (PEG). In this study, we report generation and in vitro characterization of N-terminal mono-PEGylated rSsoPox(2p) (a variant of rSsoPox(wt) having enhanced OP-hydrolyzing activity). The enzyme was PEGylated with mPEG-propionaldehyde and the PEGylated protein was isolated using ion-exchange chromatography. Compared with the unmodified enzyme, mono-PEGylation of rSsoPox results in improvement in the thermostability and protease resistance of the enzyme. PEGylated rSsoPox(2p) can be developed as a candidate for the prevention / treatment of OP-poisoning.

摘要

SsoPox是一种来自嗜热栖热菌的约35 kDa的酶，它能够水解并使多种有机磷酸酯（OP）化合物失活。该酶是开发用于预防和治疗人类OP中毒的药物的潜在候选物。然而，重组SsoPox的治疗应用存在一些与重组蛋白药物使用相关的局限性。通过将SsoPox酶与聚乙二醇（PEG）偶联，可以克服其中一些局限性。在本研究中，我们报告了N端单聚乙二醇化的rSsoPox(2p)（rSsoPox(wt)的一种具有增强的OP水解活性的变体）的产生及其体外特性。该酶用mPEG-丙醛进行聚乙二醇化，并使用离子交换色谱法分离聚乙二醇化的蛋白。与未修饰的酶相比，rSsoPox的单聚乙二醇化导致酶的热稳定性和蛋白酶抗性得到改善。聚乙二醇化的rSsoPox(2p)可开发成为预防/治疗OP中毒的候选药物。