After lipase production using vegetable oil the samples were centrifuged and applied for ammonium sulphate precipitation. But there is a problem I faced of getting oils which confused me among enzyme precipitates and oil.
Therefore, kindly any one suggests me how to separate oils from enzyme solutions without hampering the enzymes because enzymes are thermo-labile in nature.
Thanking you in advance.
Hi Abhas the problem is you will never get rld of the oll an it will always by a neusance. Tkee heart, if yuo have an thermolable emzyme ln quarntty them you don't have to enplane why you needed to receclle them
My first thought was simply to freeze out the aqueous phase, but I guess I was wrong.
Hi Abhas, you can separate the excess oil by centrifugation before ammonium sulphate precipitation since oil and water have different densities. Use little oil for lipase production since it acts as an inducer, so that the oil you adds completely used up the cells.
Thank you Dr Kavitha... Actually I used 20% oil and that's the problem may be...
By the way thanks for the suggestion... Really appreciated...
Hey Abhas all lipids have critical micellar concentrations (CMCs). This means that they all form lipid particles in aqueous solutions.
You can add a truckload of (NH3)2SO4 to your assay but it will not change the fact that lipids will still be there and can screw up your results.
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