Sometimes I need to centrifuge reactions containing enzymes, however, I've realized I have absolutely no idea if spinning very fast could damage the enzymes.
Does anyone have any data on safe centrifuge speeds for enzymes?
I imagine the most sensitive enzymes might be multi-protein complexes -- those might be easily pulled apart by high speeds.
Not really, no. They're too small to feel the g-force. You can sediment entire viruses in an ultracentrifuge at 100,000xG without killing them. They just feel a gentle pull. Individual proteins are too small to sediment at all. The machine would tear itself apart before it could ever go fast enough to affect an individual protein.
You can actually pellet proteins at ultracentrifuge speeds (which is the principle behind analytical ultracentrifugation), but generally speaking they will not be damaged. Large macromolecular complexes, such as ribosomes, may be more delicate.
Here are a few articles that describe methods for purification of ribosomes by centrifugation. They should give you a sense of tolerable g-forces.
http://cshprotocols.cshlp.org/content/2015/3/pdb.prot081331.full
Article Ribosome Purification Approaches for Studying Interactions o...
Thank you very much -- it seems clear anything below 10 xg should be tolerated easily for even large complexes, and then speeds of 10s or even 100s of xg should probably be fine too.
So 100xg is probably fine even for large complexes, but a little vortexing may completely kill an enzyme.
Is vortexing bad because of the foaming, or the shear forces?
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