I am designing protein expression in Pichia Pastoris GS115 Mut+.
I wonder which is the best way to add MeOH; either once every 24h to final 1% concentration (pulsed mode) or continually at some speed published in the literature so that cells wouldn't grow at max speed since that should improve protein yield (μ
Hi Jure,
In our lab, we've always gone with feeding either once or twice a day. For us, any differences between feeding once per day and twice per day are negligible, with regards to protein yield). However, we feed with 2% MeOH, each feed. We tend to go for a single feed, just out of ease and minimal yeast handling (thus minimising any potential contamination).
Depending on the protein in question, we get a yield of between 5 to 20 mg of protein from a 1L culture (starting volume, then down to 250-300 ml for induction phase).
All the best with your protein expression!
Thank you for your answer. Currently our bioreactor doesn't work since we need to order some parts, so I will try with baffled flask and add MeOH once a day like you do. I guess it should be fine, however it would be interested when I repeat that experiment later in bioreactor.
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