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Best method available for Yeast cell lysis
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Boopathi S
Aug 16, 2011, 4:29:02 AM8/16/11
to ye...@magpie.bio.indiana.edu
Dear Members
Please kindly provide the information about the best method available today to lyse the Yeast cell. We are using Pichia Expression system to express an thermophilic enzyme. Pichia X-33 is used for the expression. I found in some reference that the efficiency in using Glass beads for breaking the cell wall is low when compared to Yeast extraction reagents. But we cant use the Yeast Extraction Reagent due to the presence of deteregents which may affect the enzyme activity and whether the sonication is a better choice over Glass beads. Kindly help me in this regard.
Boopathi
Please kindly provide the information about the best method available today to lyse the Yeast cell. We are using Pichia Expression system to express an thermophilic enzyme. Pichia X-33 is used for the expression. I found in some reference that the efficiency in using Glass beads for breaking the cell wall is low when compared to Yeast extraction reagents. But we cant use the Yeast Extraction Reagent due to the presence of deteregents which may affect the enzyme activity and whether the sonication is a better choice over Glass beads. Kindly help me in this regard.
Boopathi
Martin Wang
Aug 16, 2011, 1:57:13 PM8/16/11
to Boopathi S, ye...@magpie.bio.indiana.edu
Hi Boopathi,
Sonication may increase the sample temperature significantly. Something you need be aware of.
In case glass bead surface interacts to protein you are working on, there are other types of beads, such as zirconium silicate, zirconium oxide, stainless steel beads. Zirconium beads have quite inert surface.
If foaming is an issue for your protein, none of the beads based method will help for they all creating bubbles during homogenization.
Martin
Martin Wang
WISBIOMED LLC
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