Hi:
we are working on isolating bacterial strains that produce secreted cholesterol oxidase using a minimum medium with cholesterol as the sole carbon source. I noticed that the cholesterol forms an emulsion and obviously doesn't like to dissolve well. So the bugs might see very little cholesterol in solution. This problem is not mentioned in the paper we follow.
The questions are: a) does that matter? and if so b) What do I do to get cholesterol dissolved? What would be a good detergent to use without killing the bugs?
I like your cyclodextrin idea - we have it in the lab. Otherwise we try with Tween as you suggested.
I have tried getting cholesterol into solution for bacterial growth, and it is possible. I too was following a method in a paper, but when I contacted the authors for some advice they admitted they could not say just how much cholesterol had gone into solution as they filtered it.
I managed to get 0,1mg/ml of cholesterol into my media and it was sufficient to see bacterial growth. I used tyloxapol and ethanol to dissolve the cholesterol and added it at 80 degrees celcius to pre-heated media. More growth was seen in the colesterol than on either tyloxapol or ethanol controls.
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