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.