Hello everyone,
I'm working on a protein, homologue of which is associated with cell wall (I don't know about my protein yet, but it should be in the cell wall as well).
I expressed this protein in Pichia pastoris under AOX1 promoter control with alpha-factor for secretion to medium. However, I could not detect any activity after 4d expression.
Is it possible that the protein would stay in the yeast cell wall and thus could not be detected in medium?
Do you know, if there is some protocol for extraction of proteins from yeast cell wall, which would not disrupt the cells? Could I use the protocol for plant cell wall-associated proteins?
Thank you.
Looking on the internet for This query's search purpose may be to determine whether a plant cell wall-associated protein can remain in the cell wall when produced in yeast, Pichia pastoris. “Plant cell wall-associated protein expression in Pichia pastoris" might be a search phrase.
Plant cell wall-associated proteins (CWAPs) are proteins located in the cell wall of plant cells that are engaged in a variety of tasks such as cell wall construction, signalling, and pathogen defence [1]. For research reasons, CWAPs can be heterologously expressed in other species, such as the yeast Pichia pastoris [2]. However, it is unknown if the expressed CWAPs would persist in P. pastoris cell walls. More studies would be required to establish whether CWAPs can be used in the future.
Sources:
1 - https://doi.org/10.3389/fpls.2019.00165
2 - https://doi.org/10.1007/978-1-4939-3572-7_8
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