Beating with sand/beads in a motorized vessel containing buffer looks much simpler to me to deal with several hundred grams of mycelium at once.
You could also think about the TCA method:
- Put the mycelium in a 50 ml tube
- Add 20 ml 25 % TCA, mix the contents of the tube thoroughly
- Pipet aliquots of 2 ml suspension in 2 ml eppendorf tubes
- Place the eppendorf tubes at -20 °C for at least a few hours (overnight is also OK)
- Thaw the eppendorf tubes on ice
- Pellet the hyphae by centrifugation (5 min, 14,000 rpm, eppendorf centrifuge, 4 °C)
- Wash the pellet twice with 80 % acetone (-20 ºC), take off as much aceton as possible
- Air dry the protein pellet at RT (this takes at least 30 min)
- Suspend the pellet in 250 microliters of 1 % SDS/0.1 M NaOH.
(It takes time to completely solubilize the pellet - at least 30 min)
- Add an equal amount of 2x gel loading buffer, immediately boil the sample for 5 min at 100 ºC
- the samples can now be analyzed via SDS-PAGE
Do you have a bead beater. It is a cryo homogenizer. You can also freeze the mycelia and then grind it in mortar pestle with liquid nitrogen in the mortar all the time.
Use HEPES at pH 7.4 or whatever you prefer, DTT, PMSF, and other protease inhibitors.
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