Gil-Garcia Marcos, Ventura Salvador
Institut de Biotecnologia i de Biomedicina and Departament de Bioquímica i Biologia Molecular, Universitat Autònoma de Barcelona, Bellaterra, Spain.
Front Bioeng Biotechnol. 2021 Aug 17;9:734068. doi: 10.3389/fbioe.2021.734068. eCollection 2021.
The production of recombinant proteins using microbial cell factories is frequently associated with the formation of inclusion bodies (IBs). These proteinaceous entities can be sometimes a reservoir of stable and active protein, might display good biocompatibility, and are produced efficiently and cost-effectively. Thus, these submicrometric particles are increasingly exploited as functional biomaterials for biotechnological and biomedical purposes. The fusion of aggregation-prone sequences to the target protein is a successful strategy to sequester soluble recombinant polypeptides into IBs. Traditionally, the use of these IB-tags results in the formation of amyloid-like scaffolds where the protein of interest is trapped. This amyloid conformation might compromise the protein's activity and be potentially cytotoxic. One promising alternative to overcome these limitations exploits the coiled-coil fold, composed of two or more α-helices and widely used by nature to create supramolecular assemblies. In this review, we summarize the state-of-the-art of functional IBs technology, focusing on the coiled-coil-assembly strategy, describing its advantages and applications, delving into future developments and necessary improvements in the field.
利用微生物细胞工厂生产重组蛋白常常与包涵体（IBs）的形成相关。这些蛋白质实体有时可能是稳定且有活性的蛋白质的储存库，可能具有良好的生物相容性，并且生产效率高、成本效益好。因此，这些亚微米级颗粒越来越多地被用作生物技术和生物医学用途的功能性生物材料。将易于聚集的序列与目标蛋白融合是将可溶性重组多肽隔离到包涵体中的一种成功策略。传统上，使用这些IB标签会导致形成淀粉样支架，目标蛋白被困在其中。这种淀粉样构象可能会损害蛋白质的活性并具有潜在的细胞毒性。一种克服这些限制的有前景的替代方法利用了卷曲螺旋结构，它由两个或更多个α螺旋组成，自然界广泛使用它来创建超分子组装体。在这篇综述中，我们总结了功能性包涵体技术的最新进展，重点关注卷曲螺旋组装策略，描述其优点和应用，深入探讨该领域的未来发展和必要改进。
