There is an enzyme which pH optima is 6.5, we want to make this enzyme working well at pH 4.5, without disturbing its catalytic efficiency for a reason of industrial fermentation process. The enzyme two two important Glutamate residues in active site, which we are thinking to change its pKa value so that it would work well at pH 4.8. Please suggest what kind of mutation we should do in the active site to attain this goal. Thank you.
Hi there,
What is the actual architecture of the active site and what is the mechanism of the reaction? I guess the reaction is based on glutamate reactivity so basically you aim at lowering the respective pK of these residues. The other issue is the substrate: is it sensitive to pH from a chemical point of view?
Try to find an homologous protein in a organism which lives at low ph. Then you can align sequences and maybe find the good one. Good luck
Are there any research papers or Patents which mention the enzyme (different source/family/superfamily) working at low pH? May get a primary clue from that. If the enzyme is of industrial importance, there are high chances that there are patent(s) around it, specially filed by big enzyme companies.
The other way is use of specialized enzyme designing softwares. That can suggest you the probable changes required and predict its effect on 3D structure of the enzyme.
hi
you can take the enzyme from fungi source, always it work at the acidic medium and tolerant the high acidity.
It is very interested view can you change the pKa , if that can done is the enzyme stile a have the activity for the substrate . You can change the source of enzyme that cane produce this enzyme in acidic culture Or Expose your organism to UV mutation then culture it in acidic medium.
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