A fusion protein designed for soluble expression, rapid purification, and enhanced stability of parasporin-2 with potential therapeutic applications

Q1 Immunology and Microbiology

Biotechnology Reports Pub Date : 2024-08-05 DOI:10.1016/j.btre.2024.e00851

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引用次数: 0

Abstract

Bacillus thuringiensis parasporin-2 (PS2Aa1 or Mpp46Aa1) selectively destroys human cancer cells, making it a promising anticancer agent. PS2Aa1 protoxin expression in Escherichia coli typically results in inclusion bodies that must be solubilized and digested by proteinase K to become active. Here, maltose-binding protein (MBP) was fused to the N-terminus of PS2Aa1, either full-length (MBP-fPS2) or truncated (MBP-tPS2), to increase soluble protein expression in E. coli and avoid solubilization and proteolytic activation. Soluble MBP-fPS2 and MBD-tPS2 proteins were produced in E. coli and purified with endotoxin levels below 1 EU/μg. MBP-fPS2 was cytotoxic against T cell leukemia MOLT-4 and Jurkat cell lines after proteinase-K digestion. However, MBP-tPS2 was cytotoxic immediately without MBP tag removal or activation. MBP-tPS2′s thermal stability also makes it appropriate for bioproduction and therapeutic applications.

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为可溶性表达、快速纯化和增强寄生虫素-2 的稳定性而设计的融合蛋白，具有潜在的治疗用途

苏云金芽孢杆菌寄生虫素-2（PS2Aa1 或 Mpp46Aa1）可选择性地破坏人类癌细胞，是一种很有前途的抗癌剂。在大肠杆菌中表达 PS2Aa1 原毒素通常会产生包涵体，这些包涵体必须经过蛋白酶 K 的溶解和消化才能具有活性。在这里，麦芽糖结合蛋白（MBP）与 PS2Aa1 的 N 端融合，无论是全长（MBP-fPS2）还是截短（MBP-tPS2），都是为了增加大肠杆菌中可溶性蛋白的表达，避免溶解和蛋白水解活化。可溶性 MBP-fPS2 和 MBD-tPS2 蛋白在大肠杆菌中产生并纯化，内毒素水平低于 1 EU/μg。经蛋白酶-K 消化后，MBP-fPS2 对 T 细胞白血病 MOLT-4 和 Jurkat 细胞株具有细胞毒性。然而，MBP-tPS2 在未去除 MBP 标记或未激活的情况下立即具有细胞毒性。MBP-tPS2 的热稳定性也使其适用于生物生产和治疗应用。

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来源期刊

Biotechnology Reports Immunology and Microbiology-Applied Microbiology and Biotechnology

CiteScore

15.80

自引率

0.00%

发文量

审稿时长

55 days

期刊介绍： Biotechnology Reports covers all aspects of Biotechnology particularly those reports that are useful and informative and that will be of value to other researchers in related fields. Biotechnology Reports loves ground breaking science, but will also accept good science that can be of use to the biotechnology community. The journal maintains a high quality peer review where submissions are considered on the basis of scientific validity and technical quality. Acceptable paper types are research articles (short or full communications), methods, mini-reviews, and commentaries in the following areas: Healthcare and pharmaceutical biotechnology Agricultural and food biotechnology Environmental biotechnology Molecular biology, cell and tissue engineering and synthetic biology Industrial biotechnology, biofuels and bioenergy Nanobiotechnology Bioinformatics & systems biology New processes and products in biotechnology, bioprocess engineering.