To understand my Bioprocessing module and better, I have been reading a lot about bacterial fermentation. I came across a lot of literature that either stuck to heat induction or biochemical induction for the expression of recombinant proteins in the form of inclusion bodies. However, I have also heard and read about processes that use a combination of both. For example, perform biochemical induction at 37 degree C and after 4-5 hours increase the temperature to 42 degree C and run the process for another 9-10 hours. In such cases where a combined approach is taken, is heat induction done to promote inclusion body formation? Does increasing temperature promote better IB formation or does it allow for more prolonged expression of the recombinant protein? Hit a dead end thinking about this and would appreciate any responses. Thank you in advance.
The process of protein-overexpression leads to celular stress. In many cases , E. coli can deal with this stress and produce the proteins in the soluble fraction. In other cases, the over-expressed protein, which may be toxic or induce cellular stress, is sent to the inclusion bodies (insoluble fraction). If you increase the temperature you are stressing the cells even further, and this may promote the formation of inclusion bodies.
A heat shock at 42 degrees will result in heat shock proteins being produced, which can help in the formation of inclusion bodies.
its a complex situation! see here Article Challenges Associated With the Formation of Recombinant Prot...
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