I want to test whether the AI-2 produced by bacteria A impact the growth of bacteria B. I want to extract AI-2 but remove proteins, sugars and others from culture broth that might also stimulate bacterial growth. The extract does not need to be pure compound, but it is better to remove proteins and sugars from broth.
Do You need sole AI-2 molecule for structural research or You need it just to check if Your bacteria produce AI-2? In case of the latest issue, it is enough to centrifuge culture and use supernatant for bioluminescence test, utizling reporter, AI-2 deficient, Vibrio strain. In case of structural studies, You should go to Your Chemistry Department with big bottle of whiskey or bunch of flowers (depending what they like:) and tell them about Your problem. Maybe they can help You. There was a good paper that You may find interesting:
Identification, Quantification, and Determination of the Absolute Configuration of the Bacterial Quorum-Sensing Signal Autoinducer-2 by Gas Chromatography–Mass
Spectrometry.
BTW, it is easier to synthesise AI-2 molecule than to isolate it from bacterial culture:)
Best,
AJ
It seems that AI-2 is also soluble in water. How about some organic solvents that might help to extract AI-2 from culture broth?
AI-2 is highly soluble in water. So are 90% of compunds You have got in the culture broth. That is the problem:). Do You want to extract AI-2 for some specific purposes?
Hi Adam, I want to test whether AI-2 is used by bacteria A to interact synergistically with other species. I want to add the cell-free supernatant of A to bacteria B to see whether supernatant A enhance growth of B in a sugar-limited medium. So I have to remove residual sugars from A before adding to B.
Dear Daniel, Analytical Chemistry Department is the place You should go to. Ask about HPLC method. It should be appropriate for Your purposes. What kind of chemical molecule Your AI-2 is? Is it borate ester or R-THMF? I am asking, becasue maybe it would be easier for You to synthesize this compound and add it to Your bacteria B?
Thank you very much, Adam. Actually, I am not sure what types of QS molecules bacteria A and B are using to communicate. I think AI-2 might be a good start because it is found common in inter-species communication. I probably will use DPD as positive control and add to bacteria B.
It's clever - You want to use precursor molecule and let Your bacteria B do the rest?
https://docksci.com/determination-of-autoinducer-2-in-biological-samples-by-high-performance-liquid-_5a98e6cfd64ab2cfb20f883b.html
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