Disruption of fungal cells

[stewart white]

Dear Michel,
            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. 
student)