Re: Get a plasmide - buy? share?

It's messy, so you're far better off buying your lysozyme from
www.brouwland.com - But if you really want to, it goes roughly like this:
1) Mix 10mls egg *white* with 30mls Vodka (assuming 40% ethanol vodka:
you are aiming for a final egg/vodka ethanol concentration of 30%)
2) Leave at room temperature for about 3 hours.
3) Dilute in water at least 50/50 to stop precipitation of further
protein from occurring.
4) Filter out all the crap using a coffee filter
5) Dry it out gently, perhaps on a wide, flat, very clean pan.
6) Collect crystallised residue, which should have lysozyme activity.

To determine how *much* lysozyme you've got, you may have to standardise
your batches by adding some in different concentrations to
just-inoculated batch cultures of a susceptible bacterium, and
standardising against the minimal inhibitory concentration. In other
words, for each batch, make five or so different dilutions of the powder
and add 1ml of each dilution to a 9ml potato-dextrose-broth culture
you've just spiked with a fresh overnight culture of B.subtilis or the like.

You could also use freshly spiked yoghurt, as both relevant cultures in
natural yoghurt are gram-positive and should be susceptible to lysozyme.

Bear in mind that while this protocol is based upon genuine literature
hunting, I've never tested the resulting lysozyme. Like the agarose
misadventure, it was a proof-of-principal thing for me: I just wanted to
know that I *could* have lysozyme if I really needed it and supply
sources dried up! :)

On 02/01/12 11:43, Mega wrote:
> Lysozyme from eggs? that sounds great!! Do you have a protocol/
> instruction about this?
>
> Do I just have to mix egg (the yellow and the white piece - both of
> them?) with ethanol and kind of 'shake' it (using a kitchen
> machine)?
> Aren't there any other proteins/enzymes that will be soluted by the
> ethanol??

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