Hi,
I am interested in extracting polar lipids from my bacterial isolate and running them on a two-dimensional TLC.
Can anyone tell me how much starting material I need? My bacterium does not grow to very high ODS and it grows on an expensive novel medium. So, I cannot grow liters of it in a broth. I can grow 100 mL and have the bacteria grow to 0.6 to 1.0 OD. Is that sufficient?
Can anyone share a protocol to extract the p-lipids from the bacterium?
Article An integrated procedure for the extraction of bacterial isop...
Hi Birikan
I am doing something similar at the moment, and I grow them in 50mL falcon tubes, with 30mL of media. I find that combining 4 of these I obtain a pellet of around 300mg. The best is to freeze-dry this pellet and then extract the lipids.
I use the Folch method to extract total lipid, but with a modification, after extracting I leave the pellet in Cl:MeOH (0.01% BHT), sealed with N2, in the cold room for few hours, before continuing with the protocol. I introduced this step after having problems to get the expected amount of lipid, and after doing some research about it. Other researchers have encountered this problem before and leaving the bacteria in the solvent longer, helps.
This would give you the total lipid, which you can easily separate into polar and neutral lipids using TLC.
I hope it helps
Noemi
Hello Bikiran Pardesi Noemi Tejera
My bacteria is Acinetobacter baumannii. I want to extracted Lipid or glycolipid . Can you give me full protocol ? Thanks.
Hi Nazmul
Your bacteria is a gram neg, so it should be easier to extract than Staph. So I would try the Folch method to start with.
Check the paper I attach here. They compared the Folch method,Bligh and Dyer method and the Chandramouli’s protocol. This last one was key for me working with Staphylococcus, because it was really hard to break and I was having very low recovery of lipids.
Let me know if you have any questions
noemi
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts