Microwave assisted solubilization of inclusion bodies

Sustainable Chemical Processes

Ishara Datta¹  Saurabh Gautam¹  Munishwar N Gupta¹ 

[1] Department of Chemistry, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India

关键词: Thioredoxin;    Maltose binding protein;    Microwave assisted reactions;    Protein refolding;    Solubilization of inclusion bodies;   

Others  :  789191
DOI  :  10.1186/2043-7129-1-2

 received in 2013-01-14, accepted in 2013-03-26,  发布年份 2013

【 摘 要 】

Background

Production of recombinant proteins in bacterial hosts often produces insoluble intracellular particles called inclusion bodies. Recovery of active protein from inclusion bodies generally requires their solubilization in chemical denaturants followed by a refolding strategy. The solubilization is carried out with shaking/stirring and takes several hours.

Results

Using inclusion bodies of seven diverse kinds of recombinant proteins [mutants of controller of cell division or death protein B (CcdB), human CD4D12, thioredoxin fusion protein (malETrx), mutants of maltose binding protein (MBP), single chain variable fragment (ScFv) b12 and single chain antigen binding fragment (ScFab) b12 (anti-HIV-1)], it is shown that exposure to microwave irradiation (200 W) for 2 min, solubilized these inclusion bodies completely. This was confirmed by data based upon turbidity measurements at 400 nm and dynamic light scattering studies. These solubilized inclusion bodies could be refolded correctly in all the cases by known methods. The refolding was confirmed by fluorescence emission spectra and biological activity studies.

Conclusion

Solubilization of the inclusion bodies before refolding is a part of protein production processes for several recombinant proteins which are overexpressed in the bacterial host systems. Our results show that microwave assistance can considerably shorten the process time.

【 授权许可】

   
2013 Datta et al.; licensee Chemistry Central Ltd.

【 图 表 】

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