[Yeast] Best method available for Yeast cell lysis

Martin Wang via yeast%40net.bio.net (by mwang from wisbiomed.com)
Tue Aug 16 12:57:13 EST 2011


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

1-866-692-1249

Office: 650-242-8368

Voice: 650-488-3685

Fax: 650-242-8244

 

mwang from wisbiomed.com

SKYPE: wisbiomed

web: www.wisbiomed.com

 

From: yeast-bounces from oat.bio.indiana.edu
[mailto:yeast-bounces from oat.bio.indiana.edu] On Behalf Of Boopathi S
Sent: Tuesday, August 16, 2011 1:29 AM
To: yeast from magpie.bio.indiana.edu
Subject: [Yeast] Best method available for Yeast cell lysis

 

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

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