Disruption of fungal cells
s.white at strath.ac.uk
Thu Oct 26 13:52:40 EST 1995
I am working on autolysis in filamentous fungi, specifically,
P.chrysogenum. We are attempting to quantify autolysis using image
analysis, and enzymology. To determine the level of enzymological activity
in the samples taken from the fermenter, we disrupt the centrifuged
biomass in a Braun Homogeniser. This requires CO2 cooling, and the
addition of glass beads. The homogeniser vibrates at approx. 2000 to 4000
rpm, disrupting the hyphae. We then centrifuge this, assaying the
supernatant for Total proteolytic activity, and glucanase activity.
Cell disruption by this method is crude and time consuming.
However, we had a loan of a cell disrupter, from constant systems in
England. This was excellent, using high pressure, forcing the sample thru
a tiny orifice (cost £4000). With regard to your method, even with the
homogeniser, and the cell disrupter, you will always see broken hyphae
etc. This is why we wash and centrifuge 2 or 3 times.
I hope this has been of some use, please feel free to contact me,
as literature in this area is not so clear (at least to me!).
Best Regards from rainy Glasgow.
Stewart White (research assisstant soon to be PhD.
More information about the Mycology