iwilson at molbiol.ox.ac.uk iwilson at molbiol.ox.ac.uk
Wed Nov 3 07:47:28 EST 1993

In article <9311011254.AA02186 at dxi.nih.gov>, lichten%bchem.dnet at DXI.NIH.GOV (Michael Lichten) writes:
> Jens Alfken asks about zymolyase and the "yeast lytic enzyme" sold by ICN.  
> The first thing to get is the ICN 1992-1993 catalog.  In it you will find a 
> more complete listing.  ICN will be happy to sell you Zymolyase (either 20T 
> or 100T) which is repackaged from the original supplier.  They also sell a 
> variety that they just call yeast lytic enzyme.  All are prepared from 
> Arthrobacter luteus and all are (presumably) the same enzyme, just with 
> different levels of activity and different levels of contaminating 
> activities (i.e. proteases, DNases, RNases, etc.).
> We find that, for routine DNA preparations, the yeast lytic enzyme (cat 
> #152270) is quite satisfactory and is by far the cheapest ($0.3 per 1K 
> units).  We use it routinely.  For more demanding purposes, such as 
> chromatin preparation, we still use zymolyase 100T (cat #32-093-1) which is 
> much more expensive ($12 per 1K units), and there are those in the 
> chromatin business who claim that you really should purify your own 
> lyticase to get rid of all the proteases.  We have no experience with 
> spheroplast transformation, so I can offer you no help on that account.
> [...]
> Michael Lichten, lichten at helix.nih.gov

Dear Michael,
	On what scale do you do this? How many grams of cells? We want a 
gentle technique of releasing microsomal enzymes. Since we get no activity in 
one of the enzymes (oligosaccharyltransferase) after French pressing (we do 
get activity of mannosyltransferases), perhaps this lytic enzyme would help. I 
would be grateful if you could e-mail a detailed protocol.

Iain Wilson
Dyson Perrins Laboratory, University of Oxford
iwilson at molbiol.ox.ac.uk

More information about the Yeast mailing list