I am evaluating the effect of my compound on pyocyanin inhibition. For that, I used different concentration of compound along with P. aeruginosa (ATCC) and it was incubated for 36 hours. I did with standard protocol for estimation of pyocyanin. I could see visually the pink colour after extraction with 0,2N HCl. I found gradual decrease of pink colour along with the increased concentration of compound. But while I am taking spectrum from 400 to 600nm, the peak started to raise at 420nm and I found maximum at 520nm. But all the values were negative. Means graph exist below 0. I used 0.2N HCl as a blank. Though the solution was intense pink colour, the readings were negative. What might be the reason.?
Hi, has anyone extracted pyocyanin from P.aeruginosa PAO1? I tried the pyocyanin extraction using the commonly used chlorofom/HCl method but did not notice any color change. The method did show a characteristic color change for one of my isolates. Literature says PAO1 produces pyocyanin. I am not sure what went wrong or is it PAO1 only produces pyocyanin when grown in any particular medium?
when i cultured it in minimal media it did not produce pyacyanin appreciably. it produces more in peptone rich medium, try that out. give atleast 48 hours incubation.
To Syed Shifat Ahmed: Same for me. When I used minimal media did not produce it. Like said previously, try with a rich medium (like GSNB or PPGAS) for at least 48 hrs (at 37C worked for me)
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