What are the drawbacks of Autoinduction of bacterial culture for protein purification? What about the stability of the protein expressed by such methods. Does anyone know about the report that protein expressed by this method got degraded after the purification?
This is totally depended on the protein you try to express. I succesfully expressed lots of proteins by now. For some proteins it worked perfectlx fine, even better than with normal expression. On the other hand for some it did not work at all.
And that's the major draw back i see, with autoinduction media. It is hard to finetune the the level of expression. With normal induction, for expamle with IPTG, you can play around with several parameters:
-concentration of IPTG:
The concentration of IPTG playes a huge role. For most proteins a standart concentration of 1mM works fine, for others it's to much and they will get insoluble. In some cases i had to go down to 0.25mM
-the OD where you start expression
-the temperature you express your culture at
I usually express proteins at a lower temperature than 37°C (in some cases down to 13°C). With autoinduction media, you can do this as well, but your cells will take much longer to grow
I never had reason to believe my proteins degraded, because of a certain method. But maybe someone knows more about that.
In my opinion, just try if it works for yourself. And good luck!
Cheers
Simone answered it quite very well. It totally depends on the protein which you want to express, for example in some cases the expressed protein might be toxic to cells so you may want to be able to induce the expression at a specific time and for a specific interval. Other than that, the previous answer is very well put.
Best
I have good experience with autoinducing medium (protein yield and solubility) and it worked best if I grew E.coli @37°C to a OD of about 0.3-0.6 (this is when protein expression is typically induced after consumption of glucose from the medium) and then continue shaking at 16-20°C over night for protein expression. But of course it very much depends on the protein itself what strategy for expression and purification works best.
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