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热诱导型表达系统在大肠杆菌中生产重组蛋白:进展与见解。

Thermoinducible expression system for producing recombinant proteins in Escherichia coli: advances and insights.

机构信息

Unidad de Bioprocesos, Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, CP 04510, Ciudad de México, México.

Probiomed S.A. de C.V. Planta Tenancingo, Cruce de Carreteras Acatzingo-Zumpahuacan SN, 52400 Tenancingo, Estado de México, México.

出版信息

FEMS Microbiol Rev. 2021 Nov 23;45(6). doi: 10.1093/femsre/fuab023.

Abstract

Recombinant protein (RP) production from Escherichia coli has been extensively studied to find strategies for increasing product yields. The thermoinducible expression system is commonly employed at the industrial level to produce various RPs, which avoids the addition of chemical inducers, thus minimizing contamination risks. Multiple aspects of the molecular origin and biotechnological uses of its regulatory elements (pL/pR promoters and cI857 thermolabile repressor) derived from bacteriophage λ provide knowledge to improve the bioprocesses using this system. Here, we discuss the main aspects of the potential use of the λpL/pR-cI857 thermoinducible system for RP production in E. coli, focusing on the approaches of investigations that have contributed to the advancement of this expression system. Metabolic and physiological changes that occur in the host cells caused by heat stress and RP overproduction are also described. Therefore, the current scenario and the future applications of systems that use heat to induce RP production are discussed to understand the relationship between the activation of the bacterial heat shock response, RP accumulation and its possible aggregation to form inclusion bodies.

摘要

大肠杆菌中重组蛋白(RP)的生产已经得到了广泛的研究,以寻找提高产物产量的策略。在工业水平上,通常采用热诱导表达系统来生产各种 RP,从而避免了化学诱导剂的添加,从而最大限度地降低了污染风险。噬菌体 λ 的调控元件(pL/pR 启动子和 cI857 热敏阻遏物)的分子起源和生物技术用途的多个方面为改进使用该系统的生物工艺提供了知识。在这里,我们讨论了 λpL/pR-cI857 热诱导系统在大肠杆菌中生产 RP 的潜在用途的主要方面,重点介绍了有助于该表达系统发展的研究方法。还描述了宿主细胞因热应激和 RP 过度产生而发生的代谢和生理变化。因此,讨论了使用热诱导 RP 生产的系统的当前情况和未来应用,以了解细菌热休克反应的激活、RP 积累及其可能聚集形成包涵体之间的关系。

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