Hello,
I need help to purify my product benzyl butyrate from my lipase-katalyzed esterification. I am using Novozym ® 435 and in addition molecular sieves. I want to quantify the amount of product with GC analytic but I don‘t know how to remove the enzyme and the sieves.
I tried filtration with several wash steps with aceton but I am not able to get all the product and the remaining substrates out of my reaction vessel. Furthermore I don‘t know how to separate the enzyme and the sieves efficiently.
Thank you in advance for your help :)
How about extracting the mixture with a non-miscible organic solvent, such as ether ? In the old days, I would have suggested benzene, but nowadays it raises too many safety problems, being a class 1 carcinogen.
Pierre Béguin thank you!
Any chance you know how to separate the enzyme from the molecular sieves?
Considering solubility of the reagents and the product, it is best to use solvent like methyl isobutyl ketone (MIBK) or 2-methyl tetrahydrofuran. The Novozyme 435 activity is not affected.You could probably use 10 volumes of solvent (5+5) to wash the beads. The molecular sieves would be fine. You can simply decant the upper organic layer and reuse the mixture of enzyme beads and mol.sieves.
It would be difficult to separate the molecular sieves from the enzyme beads. We generally use them in a separate tea-bag hanging in the reactor.
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