Does anyone know how to inexpensively obtain gluconacetobacter xylinus?

[Philipp Kollenz]

Hi there

I am trying to produce nanocellulose and the cultures you can buy on websites like this one (https://www.dsmz.de/catalogues/details/culture/DSM-2325.html) are too expensive. I heard it is part of a microbe mix that is used to make some kind of asiatic tea beverage called Kombucha. Is it possible to isolate it from there?

best regards,

Phil

[Sebastian S Cocioba]

Either from the tea or contact Kombucha in brooklyn and get a mother for $20 something USD. 

Sebastian S. Cocioba

CEO & Founder

New York Botanics, LLC

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[Sebastian S Cocioba]

http://www.kombuchabrooklyn.com/supplies/cultures/kbbk-scoby-371.html

Sebastian S. Cocioba

CEO & Founder

New York Botanics, LLC

On Jun 7, 2015, at 7:37 AM, Philipp Kollenz <philipp...@googlemail.com> wrote:

--

[Cathal (Phone)]

It's easy to isolate from Kombucha, and some literature suggests this strain gives the best cellulose, too. :)

Just make up "Gluconobacter agar" (look it up on thelabrat.com) and streak your kombucha culture over it thinly. When I isolated it in this way, my colonies were brown, cleared the CaCO3 quickly, and in *late* culture, white crystals (of cellulose?) appeared around, but rarely atop, colonies.

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[BraveScience]

Hi guys,

At the OpenWetLab in Amsterdam we did something similar to what Cathal suggested above.

 

Using calcium carbonate helps indeed isolating acetic acid producing strain, but as pH of the medium goes up, you will see pretty much the zygomyces taking over. You can spot them by their white colonies that create sort of a "bump" on the agar plate.

G.xylinus colonies are brownish also in my case. But I could spot them (after 3-4 days at 30°C) just if I added EtOH to the medium.

My suggestion is to add 4-6% alcohol to the medium, this will slow down yeasts and speed up Gluconacetobacter metabolism.

Be careful to streak the isolated colonies at least 3-4 times. Cleaning the strain from other microorganisms isn't always straightforward.

Best,

Fede

[Philipp Kollenz]

I have two different types of colonies: 

- Flat and brownish

- "Bumpy", glossy, and almost clear

They both aren't yeasts (as far as I could tell with my microscope) and grew after 5-6 days at 26°C 

Which of them is the one I'm looking for?

[Cathal Garvey]

IIRC (it was a while ago) G.xylinus is a dark brown and has little
distinctive shape; flat and brownish sounds about right. The
glossy/clear partner sounds like it might be a yeast.

Take pickings from distinct, separate examples of both colonies and
streak them out again on separate agar, you'd expect the brown guys to
*prodigiously* clear the CaCO3 in the agar and make it
clear/see-through, and in late-stage culture (+7 days or even later) to
see chunks of cellulose forming near, though not necessarily *on*,
colonies. That was my experience, at least.

As for the white guys, check 'em under a microscope; if they're yeast,
they're probably much larger than bacteria and easy to observe, for
starters. You might even see nuclei without a stain, though try
methylene blue for a cheap, cheerful DNA stain.

On 01/07/15 20:47, Philipp Kollenz wrote:
> I have two different types of colonies:
> - Flat and brownish
> - "Bumpy", glossy, and almost clear
>
> They both aren't yeasts (as far as I could tell with my microscope) and
> grew after 5-6 days at 26°C
> Which of them is the one I'm looking for?
>
> Am Sonntag, 7. Juni 2015 19:33:09 UTC+2 schrieb Cathal (Phone):
>
> It's easy to isolate from Kombucha, and some literature suggests
> this strain gives the best cellulose, too. :)
>
> Just make up "Gluconobacter agar" (look it up on thelabrat.com

> <http://thelabrat.com>) and streak your kombucha culture over it

> thinly. When I isolated it in this way, my colonies were brown,
> cleared the CaCO3 quickly, and in *late* culture, white crystals (of
> cellulose?) appeared around, but rarely atop, colonies.
>
> On 7 June 2015 12:37:54 GMT+01:00, Philipp Kollenz

> <philipp...@googlemail.com <javascript:>> wrote:
>
> Hi there
>
> I am trying to produce nanocellulose and the cultures you can
> buy on websites like this one
> (https://www.dsmz.de/catalogues/details/culture/DSM-2325.html

> <https://www.dsmz.de/catalogues/details/culture/DSM-2325.html>)

> are too expensive. I heard it is part of a microbe mix that is
> used to make some kind of asiatic tea beverage called Kombucha.
> Is it possible to isolate it from there?
>
> best regards,
> Phil
>
>
> --
> Sent from my Android device with K-9 Mail. Please excuse my brevity.
>

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[Patrik D'haeseleer]

Any recommendations for a good antifungal that could be used in such situations? Something that's going to inhibit all the yeast and molds from growing, but won't affect the bacteria.

I keep wondering if some of the usual athlete's foot / jock itch products can be used for this, as an easily accessible DIY solution...

Patrik

[Cathal (Phone)]

Huh, interesting thought. As long as they aren't also medicated for secondary bacterial infection (as some are)?

The powders are mostly talc, so unlikely to affect the agar formula much. Would need to be added after sterilising though, and are topical (ie designed for unbroken skin) formulas guaranteed sterile if unopened?

[Theresa Ramseyer]

What is keeping you from getting a bottle of one of the organic
versions of the tea and using it? Seems that would have antifungal
properties from being bottled. Does the bottling process kill off what
you're looking for?

(Probably a very dumb question, but I know/remember very little
biology. The subject intriques me. I'm on
this list to learn. )

Theresa

[Nathan McCorkle]

Not a bad idea, you'd need to get rid of any cells floating around. Maybe centrifuge hard, and then maybe even filter with something like a 0.2 micron filter.

[Patrik D'haeseleer]

Actually, many of the commercial "kombucha" brands that I've seen sold in stores are NOT actually made with G. xylinus. 

As I understand, natural kombucha cultures have a problem with too much pressure buildup after bottling. You can safely bottle ferment beer and champagne, because as the pressure in the bottle increases, additional CO2 production becomes less thermodynamically favorable, and eventually fermentation comes to a halt. I guess in a kombucha culture, that equilibriumis reached at a higher pressure than you would get with brewer's yeast, so you'd get exploding bottles on the store shelves. (Just speculating here, mind you.)

Patrik

[Mark Jones]

Would anyone know where I could purchase the kombucha yeasts and bacteria on a fairly large scale? As far as I know it 's the Saccharomyces and the Gluconacetobacter xylinus if thia woulod be correct?

:)