Isolating penicillin ?

[Open BioLab Graz - Austria]

Heyo,

We would like to produce our own penicillin from mold, does anyone know a simple method to isolate the penicillin? (We could use a FPLC or a capillary electrophoresis too, but i'd prefer to do it without big machines)

[Mega]

Hi...
As far as I remember, Penicillium also produces Mycotixins. If that is true, better din't burn your fingers by playing with a potential pathogene. Why not usolate e.g. Streptomycin from some kind of actinomycetes?

[Mega]

By the way, better try producing EPO... My biotechnology professor always mentions thr price of EPO being 300-500 million euros per kilogram :D

It should be legal to produce, but you are not allowed to sell it as a medicine.

[Koeng]

Mother of god

[Dakota Hamill]

I tried a ghetto liquid culture technique
http://basementbiotech.org/penicillium-isolation-from-citrus-fruit/
with inconclusive results.

Isolating a strain of pencillium is pretty easy, just let some citrus
fruit rot for a little and look for the olive green colored mould that
arises. You can make a simple microscope slide as shown, and
pencillium has very unique fan-like conidia. To make isolation of
the secondary metabolites (of which penicillin should be one) easier,
culturing the strain in a liquid culture is advised. I just boiled
some lemon chunks for a while and threw them in coffee cups and seeded
the liquid cultures with mycelium / spore swabs off the rotting citrus
fruit.

You could try some different media, there are tons of papers out there
about pencillium cultivation. I have some growing in potato dextrose
broth at the moment and it seems to be growing fine.

The reason the experiment ended in inconclusive results is that I
waited 3-4 days to check the disc diffusion assays, enough time for
the gram positive bacteria to grow back over its fallen comrades into
any area once made clear by the diffused penicillin.

I generally do DCM workups of fungal fermentation liquid media, save
and concentrate both the organic and aqueous partition, and will
sometimes macerate the mycelial mat and do an MeOH extraction. Ethyl
acetate is also a good organic solvent choice, probably better than
DCM.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1258127/pdf/biochemj00965-0101.pdf

That's an old paper but, sometimes old papers can use more simple
equipment, since fancy stuff wasn't available yet, though the catch-22
is using solvents and reagents most people would try to avoid now a
days.

I'd say, isolate some pencillium and start some pure cultures on a few
different liquid media types, then do some more reading while it grows
(around 10-12 days is when I believe optimal penicillin conc. is
reached).

Liquid liquid extraction with a separatory funnel would be pretty
easy, then you could do minimum inhibitory concentration tests by eye,
or with a spec at OD600, or disc diffusion.

[Mega]

Won't it be difficult to isolate it? I was told that wild-type strains only produce 1/10 the penicilline as bred strains and <1/1000 as engineered strains.

I don't clearly remember the numbers, it could also be 1/10,000 the amount.

But that will be quite low amounts, I assume.

[leaking pen]

those strains are for mass production at the factory level. concentrations in wild were quite succesfully isolated before!

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[Open BioLab Graz - Austria]

It's more about having some oldschool fun like the good ol' Fleming, than producing tons of penicillin :P

[Dakota Hamill]

One of the original industrial strains (perhaps still used to this day I had read) was isolated from a grapefruit found in a fruit market somewhere in Ohio or Idaho or something...so you never know what nature might give you or where it will show up!  Streptomycin was found from a bacterial strain taken from a dirt sample behind the barn of the principle investigator  credited with its discovery by one of his graduate students.  Rapamycin was found in a bacteria from a dirt sample taken on the edge of an old volcano on Easter Island....so, natural products are everywhere!