González-Ponce Karen Stephania, Celaya-Herrera Samuel, Mendoza-Acosta María Fernanda, Casados-Vázquez Luz Edith
Departamento de Alimentos. División de Ciencias de la Vida, Campus Irapuato-Salamanca, Universidad de Guanajuato, Carretera Irapuato Silao km 9, 36500, Irapuato, Guanajuato, México.
Departamento de Formación Integral e Institucional, Fraccionamiento Industrial Puerto Interior, Unidad Profesional Interdisciplinaria de Ingeniería Campus Guanajuato, Instituto Politécnico Nacional. Avenida Mineral de Valenciana 200, C.P. 36275, Silao de La Victoria, Guanajuato, México.
J Membr Biol. 2025 Feb;258(1):15-28. doi: 10.1007/s00232-024-00333-0. Epub 2025 Jan 6.
The Cell-Free Protein Synthesis (CFPS) is an innovative technique used to produce various proteins. It has several advantages, including short expression times, no strain engineering is required, and toxic proteins such as membrane proteins can be produced. However, the most important advantage is that it eliminates the need for a living cell as a production system. Membrane proteins (MPs) are difficult to express in heterologous strains such as Escherichia coli. Modified strains must be used, and sometimes the strain produces them as inclusion bodies, which makes purification difficult. CFPS can avoid the problem of toxicity and, with the use of additives, allows the production of folded and functional membrane proteins. In this review, we focus on describing what cell-free systems are. We address the advantages and disadvantages of the different organisms that can be used to obtain cell extracts, including PURE systems, where the components are obtained recombinantly, and the methodologies that allow the synthesis of membrane proteins in cell-free systems, which, given their hydrophobic nature, require additives for their correct folding.
无细胞蛋白质合成(CFPS)是一种用于生产各种蛋白质的创新技术。它具有几个优点,包括表达时间短、无需菌株工程改造,并且可以生产诸如膜蛋白等有毒蛋白质。然而,最重要的优点是它消除了将活细胞用作生产系统的需求。膜蛋白(MPs)在诸如大肠杆菌等异源菌株中难以表达。必须使用经过改造的菌株,有时该菌株会将它们产生为包涵体,这使得纯化变得困难。CFPS可以避免毒性问题,并且通过使用添加剂,可以生产出折叠且具有功能的膜蛋白。在这篇综述中,我们着重描述无细胞系统是什么。我们阐述了可用于获取细胞提取物的不同生物体的优缺点,包括通过重组方式获得组分的PURE系统,以及允许在无细胞系统中合成膜蛋白的方法,鉴于膜蛋白的疏水性,其正确折叠需要添加剂。