Singh Anupam, Upadhyay Vaibhav, Upadhyay Arun Kumar, Singh Surinder Mohan, Panda Amulya Kumar
Product Development Cell, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, 110067, India.
Microb Cell Fact. 2015 Mar 25;14:41. doi: 10.1186/s12934-015-0222-8.
Formation of inclusion bodies in bacterial hosts poses a major challenge for large scale recovery of bioactive proteins. The process of obtaining bioactive protein from inclusion bodies is labor intensive and the yields of recombinant protein are often low. Here we review the developments in the field that are targeted at improving the yield, as well as quality of the recombinant protein by optimizing the individual steps of the process, especially solubilization of the inclusion bodies and refolding of the solubilized protein. Mild solubilization methods have been discussed which are based on the understanding of the fact that protein molecules in inclusion body aggregates have native-like structure. These methods solubilize the inclusion body aggregates while preserving the native-like protein structure. Subsequent protein refolding and purification results in high recovery of bioactive protein. Other parameters which influence the overall recovery of bioactive protein from inclusion bodies have also been discussed. A schematic model describing the utility of mild solubilization methods for high throughput recovery of bioactive protein has also been presented.
在细菌宿主中形成包涵体对大规模回收生物活性蛋白构成了重大挑战。从包涵体中获得生物活性蛋白的过程劳动强度大，而且重组蛋白的产量往往很低。在这里，我们回顾该领域的进展，这些进展旨在通过优化该过程的各个步骤，特别是包涵体的溶解和溶解蛋白的重折叠，来提高重组蛋白的产量和质量。我们讨论了温和的溶解方法，这些方法基于对包涵体聚集体中的蛋白质分子具有类似天然结构这一事实的理解。这些方法在保留类似天然蛋白质结构的同时溶解包涵体聚集体。随后的蛋白质重折叠和纯化导致生物活性蛋白的高回收率。还讨论了影响从包涵体中整体回收生物活性蛋白的其他参数。还提出了一个描述温和溶解方法用于高通量回收生物活性蛋白的示意图模型。
