Practice question

Lowering post-induction temperature to 16-25°C markedly improves solubility of recombinant proteins expressed in Escherichia coli. Reduced temperature slows transcription and translation rates, decreasing nascent chain concentration and allowing more time for co-translational folding assisted by chaperones GroEL and DnaK. Hydrophobic interactions driving aggregation are temperature dependent and weakened at lower temperatures, thus inclusion body formation is minimized. High IPTG or high temperature exacerbates aggregation, while glucose suppresses induction via catabolite repression but does not enhance folding quality of already induced protein.