I was checking the heat tolerance of bacteriocin (from K. pneumoniae) and the same suspension was heated at different temperatures 30C, 40C, upto 80C, for 10 minutes each.
I did this procedure so to remove other proteins from the suspension but is it possible that bacteriocin are also degraded?
Yes, nonetheless with your experiments you will be able to determine how stable or labile your toxin is upon heating. If the toxin is small it should be more stable.
All proteins and peptides are destroyed by heat, but they differ in the temperature and the time they can withstand. Thus, heat-resistant proteins can be separated from heat-sensitive ones. First thing you need is a good assay for the activity you wish to purify, which should be sensitive, reliable, cheap and quick (as you will perform it many times).
Then you incubate samples of your source at different temperatures for different times, carefully establishing conditions that your activity is just able to survive. It is important to do this reproducibly: heat the sample in a boiling waterbath in small aliquots in vessels of high heat conductivity with constant shaking until they have reached target temperature, then transfer to a second waterbath set to the temperature under study. Keep them for the desired time, than rapidly chill in an ice/water mixture. Spin down precipitated proteins, then check the supernatant for activity.
In some cases, it is possible to increase the specific activity (activity units per mg of protein) by a factor of 3-4.
Yes the purpose of heating the bacteriocin sample was to check if they are heat labile and to what temperature. I heated the samples in water bath at various temperatures, one sample was good, I was able to isolate and identify but in the other bacteriocn sample, I think I degraded the protein completely during the heating process. Thank you so much, the answers are very helpful.
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