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Bacterial cultures help needed

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Limpet Chicken:
Hi.

Just recently, I have been giving some serious thought into culturing some bacteria, pseudomonas in particular, to harvest a particular enzyme from them.

Pseudomonas I know are commonly found growing on old bath sponges/loofahs etc. and are known for causing problems when scratched into the skin by brushes or loofahs, has anyone got any details of how to identify pseudomonas bacteria, and pictures they could upload?

Also, I am very inexperienced with biological cultures, could some kind preson perhaps guide me through a good process to grow these bacteria, I have no fancy equipment, but making up nutrient solutions with multivitamin pills, minerals etc. and sterililizing solutions is no problem.

I aim for quite a large colony, so as to extract the enzyme out of the bacteria,
if possible, I want to be able to use a two-liter coke bottle as a growing chamber, to ensure a large crop of bacteria, petri dishes are too small by far, and I only have one (I "liberated" it from school about three years ago ;D)

Limpet Chicken:
I really could use a little help with this, I need to know what sort of nutrients pseudomonas sp. organisms thrive in, from what I know its warm and wet (sounds wrong doesn't it ;)) but I have no idea what sort of stuff it likes to feed off.

How easy would it be to produce a crude extract of the bacteria to obtain the enzyme I need? biotechnology/biology has never really been my speciality, would something such as penicilin to hydrolyse bacterial cell walls then subsequent filtration through active carbon possibly prove sufficient to gather my enzyme?

Mr Amino:
Typically cell walls are broken open by a type of pressure device...i think its called a french press.  I used to use one to harvest cytochrome c from bio-engineered E. coli.  What enzyme are you trying to obtain? Keep in mind that pseudomonas is a pathogenic bacteria and it is not typically grown to harvest bio-molecules for that reason.

limpet chicken:
Pseudomonas bacteria produce an enzyme that can "bee" used to produce indole-3-carboxaldehyde.

Perhaps the pseudoname "Hest" might mean something to you (no it isn't SWIM's), and something to do with indole-3-carboxaldehyde and a microwave :)

savoy7:

--- Quote from: Mr Amino on January 19, 2005, 10:40:41 PM ---Typically cell walls are broken open by a type of pressure device...i think its called a french press.  I used to use one to harvest cytochrome c from bio-engineered E. coli.  What enzyme are you trying to obtain? Keep in mind that pseudomonas is a pathogenic bacteria and it is not typically grown to harvest bio-molecules for that reason.

--- End quote ---


to reiterate - pseudomonas is an opportunistic pathogen - if it has the opportunity, it will cause infection - like in burn victims -

I would look at the initial incubating system that you are planning.  2 liter coke bottles are nice, but making sure they are sterile would be a little difficult.  Plus - growing that stuff where one doesn't have access to L-F-hoods, lack of equipment might be hard.  Hopefully, I did dash your idea, just wanted to give you some things to consider.

savoy

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