Re: [DIYbio] Re: Kombucha as-is or isolated cultures for gel electrophoresis or other DIYbio uses

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@Nathan
I wanted a pure line of Xylinus so I could mix it with a lab strain of
yeast and have a "lab strain" kombucha. It wouldn't be as robust or as
good for brewing, probably, but might be interesting as a
kitchen-friendly "model organism"! :)

As to antibiotics, there's loads of supposition out there that, in
addition to the acid production, one or more of the species in
kombucha SCOBYs produce bacteriocins or other antibiotics. You could
test this by doing an exclusion assay, of course! Filter-sterilise
some freshly brewed kombucha (or wait til it's a bit stale to check
for stationary phase secretion), soak some sterile discs of filter
paper in that and normal boiled (and filter sterilised) vinegar, and
apply the two discs to separate freshly streaked agar plates of a test
species, like E.coli.

Preferably pick a few test species from different genuses, as
bacteriocins are rarely broad spectrum.

Compare growth exclusion radius, if any, to establish whether there's
cause to suspect more than just the pH is to blame (would want to
ensure by dilution that the pH of either sample is the same, and the
buffering capacity of the agar is the same; same batch for e.g)

@Patrik:
Spider silk requires spinning to form polymers, whether by
electrospinning or spinneret, but glue proteins or chitin sound like
nice leads!
Another thought is: I found some patents for cellulose binding
peptides, and separate patents for calcium carbonate-binding peptides.
Possibly there was a compromise patent involving combining the two to
make cheap bio-bleach for paper industry. Can't recall!
But, there were libraries in each patent of artificially-evolved small
peptides for binding to calcium carbonate and cellulose; could
conjugate those to one another to make cellulose/calcium adaptors to
perhaps help precipitate calcium carbonate into a crust over the
cellulose fibers.

If it worked, you'd probably end up with something that looked a bit
like spongy coral, provided enough CaCl to build up a sufficient crust..

On 02/26/2013 12:33 PM, Patrik D'haeseleer wrote:
> You know, I usually try to make things *easier* to degrade, but
> what if you could engineer G.xylinus to produce something a bit
> stronger than amorphous cellulose?
>
> Add an acetyltransferase and get it to produce chitin?
>
> Mix in some bioplastics?
>
> Try to secrete silk, spidersilk, or mussel glue proteins, possibly
> modified with some cellulose binding modules?
>
> Patrik
>
> On Monday, February 25, 2013 5:49:27 PM UTC-8, Nathan McCorkle
> wrote:
>
> I just got a kombucha SCOBY in the mail... I haven't brewed
> kombucha for years and the vinegary smell out of the box is a bit
> unpleasant (I know brew doesn't need to be so acidic). Other than
> drinking and electrophoresis gel+buffer, what other uses can we
> think of?
>
> Cathal mentioned isolating G.xylinus, presumably in the line of
> thinking that we could scale up production and have a nice pure
> cellulose/other-ose producer for various uses.
>
> What other ideas are there for kombucha as-is or as isolates? For
> instance, I knew a guy who grafted a bunch of cacti, then applied
> blended SCOBY to the fresh graft wounds. He said the cacti all
> healed with no failures, and thought the SCOBY may have added some
> antibiotic factor (or at least antibiotic for plant diseases).
>
> On Wed, Jan 30, 2013 at 4:44 AM, Cathal Garvey
> <cathal...@cathalgarvey.me <javascript:>> wrote:
>> Sadly I let my Kombucha stale in the fridge for too long and
>> couldn't recover it. And, my isolated culture of G.xylinus is
>> long dried
> out too,
>> due to a gap in the parafilm. So, I'll have to re-order Kombucha
>> and re-isolate G.xylinus! Thankfully, the latter isn't too
>> challenging.
>>
>> You make agar containing calcium carbonate powder, stir and pour
> right
>> before solidifying so you get evenly dispersed CaCO4 particles
> through
>> the gel, then streak and re-streak your kombucha on the surface.
>> The colonies that create a broad area of clear agar (acid
>> dissolving the CaC04) are your Geo/Acetobacter, of which there
>> may be one or two
> species.
>>
>> Isolate those and grow to stationary, the plates that form
>> nodules of whitish cellulose after a week or so are your guys.
>> :)
>>
>> I have the agar recipe lying around somewhere, will dig up soon.
>>
>> On 30/01/13 06:06, Patrik D'haeseleer wrote:
>>> Hey Cathal - do you have some Kombucha going right now? I'd
>>> love
> to see
>>> what happens if you take a nice thick "mother" SCOBY, cut some
>>> wells into it, and use it as an electrophoresis gel. :-). Food
>>> dyes electrophorese pretty well.
>>>
>>> Patrik
>>>
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