I'm studying about recombinant protein production now. When producing recombinant protein in large scale using E. coli, there are several problems including endotoxin and bad smell etc. I want to know how to solve these problems. Thank you.
E. coli smells, but you get used to it. Besides, there is much worse available in each and every household.
The smell is no safety issue. Unless you swallow a larger amount of the E.coli suspension (this happens easily when you try to pipette with your mouth - make sure not to do it!! Neither the pipetting with your mouth nor the swallowing of E. coli!!), handling the bacterium is safe, assuming it is from a known type collection and not something isolated from the wild.
If you swallow some of it, you will likely have a couple days of diarrhea. I know of no case, in which it got worse than that. But, always safety first - try not to do it and wear protective clothing, gloves, etc.
LPS can (usually) be removed in purification; depending on the protein, it may need some tinkering. I once dealt with a class of proteins that actively binds LPS - even those we got clean.
Proteins purified from E.coli culture, and in fact all pharmaceutical products intended for parenteral administration, must be tested for endotoxin. This test can be outsourced to specialized companies.
Hi Garshab,
One of the most odorous compounds from E. coli is indole which is derived from the amino acid tryptophan. What about reducing the protein/tryptophan content of the media or work with a mutant with no indole producing capability? Replace with inorganic nitrogen.
They are also more likely to produce odour is the system is not fully aerobic. This could tie in with the above, use nitrate because it can also act as an electron acceptor as well as an nitrogen source.
Andrew
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