I would like to know the culture collections or other private institutions who could provide yeast strains for protein production possessing strong promoters, selection markers and possessing other properties of a strong expression system.
And also the names of the commercial Saccharomyces expression systems.
Thanks!
I am not aware of commercially available strains with optimized properties. What you should use is protease-deficient strains, these will more likely produce your protein in a full-length form. look for strains with a mutation in the master protease Pep4. In pep4 mutants, many other proteases (those that are activated by Pep4) will also have reduced activity.
There are however commercially available systems for protein expression in the yeast Pichia which is widely used for high level production. You might wish to look into Pichia instead of Saccharomyces.
I don’t think these are available for free, they tend to be company IP and they usually require payment for their use. Companies like Ingenza offer a protein production service with shared IP, they normally charge about £10,000 per protein for the initial cloning and validation.
proteases are a problem and can be knocked out if needed but a protease free yeast strain I doubt exists. You can do some bioinformatics which highlight protease sites on your protein and that would be your first port of call.
Hope this helps?
Hi Darren,
sure, there is no protease-FREE strain. Proteases have many important biological functions. This is why the strains are called protease-deficient. I have been working with a strain that had 4 mutations: pra1-1 (synonym of pep4), prb1-1, prc1-1, cps1-3. It did not require any special treatment, had normal doubling time and was easy to use. Use of strains like this one is a safety measure.
Jürgen
Your right of course, we used to use a Pep4 ko for protein expression. We also knocked out Kex1 and kex2 when I worked in industry, interesting you have a multi protease gene knockout and it still has normal functionality. Thanks for sharing
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