Gad Shubhada, Ayakar Sonal
Department of Biotechnology, Institute of Chemical Technology - IndianOil Odisha Campus Bhubaneswar, Odisha 751013, India.
Biotechnol Rep (Amst). 2021 Sep 6;32:e00670. doi: 10.1016/j.btre.2021.e00670. eCollection 2021 Dec.
The synthesis of complex molecules using multiple enzymes simultaneously in one reaction vessel has rapidly emerged as a new frontier in the field of bioprocess technology. However, operating different enzymes together in a single vessel limits their operational performance which needs to be addressed. With this respect, scaffolding proteins play an immense role in bringing different enzymes together in a specific manner. The scaffolding improves the catalytic performance, enzyme stability and provides an optimal micro-environment for biochemical reactions. This review describes the components of protein scaffolds, different ways of constructing a protein scaffold-based multi-enzyme complex, and their effects on enzyme kinetics. Moreover, different conjugation strategies viz; dockerin-cohesin interaction, SpyTag-SpyCatcher system, peptide linker-based ligation, affibody, and sortase-mediated ligation are discussed in detail. Various analytical and characterization tools that have enabled the development of these scaffolding strategies are also reviewed. Such mega-enzyme complexes promise wider applications in the field of biotechnology and bioengineering.
在一个反应容器中同时使用多种酶合成复杂分子，已迅速成为生物工艺技术领域的一个新前沿。然而，在单个容器中同时操作不同的酶会限制它们的操作性能，这是需要解决的问题。在这方面，支架蛋白在以特定方式将不同的酶聚集在一起方面发挥着巨大作用。这种支架作用提高了催化性能、酶的稳定性，并为生化反应提供了一个最佳的微环境。本文综述了蛋白质支架的组成部分、构建基于蛋白质支架的多酶复合物的不同方法及其对酶动力学的影响。此外，还详细讨论了不同的缀合策略，即；dockerin-黏连蛋白相互作用、SpyTag-SpyCatcher系统、基于肽接头的连接、亲和体和分选酶介导的连接。还综述了推动这些支架策略发展的各种分析和表征工具。这种巨型酶复合物有望在生物技术和生物工程领域得到更广泛的应用。
