How much methanol should be added to the fermenter in which we cultivate Pichia pastoris for the purpose of heterologous enzyme production? We will add methanol continuously with the pump x ul/min.
Thanks for responses
Methanol is added at a concentration of 0.5–1.0% (v/v) to induce expression of the target protein in Pichia pastoris.
That will definitely depend on size of your bioreactor (you're not fermenting;). Also, in the beginning, the yeast will require less methanol than later, when it will be grown alot. One more thing - usually you grow the yeast on glycerol or something else in the beginning and start inducing after some time. So, you can slowly decrease the feeding of the first carbon source and increase feeding with methanol.
You can start with some small flowrate and increase it over time. You should check oxygen level. If it will spike once you turn off the methanol, you're adding too little or about the right amount. If it will not spike, than you have some left over methanol in your medium and should decrease the flowrate.
That's funny that you have strain BG11. We use medium BG11 for microalgae :)
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