I know the usual thing to do would be to induce expression for each recombinant on different batches, and collect/purify the proteins of interest separately, but has someone done this experiment before?
You can, but it's tricky to get it just right if you want the proteins to be expressed in certain amounts.
Any reason that you'd want to express both at the same time? It's super cheap and easy to have 2 flasks instead of one.
Hi, Katie A S Burnette thank you for answering. My initial experiment was to produce a single pichia that had both genes and generate the recombinant enzymatic reaction in situ (inducing expression in a medium that has the enzyme substrate). I had many issues producing this single recombinant with two genes, so I wanted to try the same thing but with two different pichia in a single medium, but I don't know if it's worth it nor am I sure if it is the correct way to approach the investigation. If it were possible to do, I also don't know how to correctly induce the two pichia in a single medium, since I should take into account elements such as standardizing the inoculum, adding more methanol, etc., and I can't find literature where someone has done this because the usual thing to do is to use a single flask for a single yeast. Thank you for reading :)
Whether it will work in your case is going to depend upon the product of the first reaction, is that product going to be excreted out of the cell in sufficient quantities for the second enzyme in the second strain to act on it. So it is hard to predict. I am sure it will work to some degree, but it may not be efficient.
There may also be a growth rate difference between the two strains. You could consider growing them separately until they reach a good density, then pool the two cultures into one flask and induce for expression.
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