The lipase enzyme should be soluble in aqueous buffer, but perhaps not at quite so high a concentration. Try making a 0.1 mg/ml solution. Care should be used when vortexing or magnetic stirring to avoid denaturing the protein. Mix gently so as to avoid creating foam.

Dear Ajay,

 Mixtures of hydrophilic organic solvent and PBS buffer such as glycerol/PBS,  ethanol/PBS, DMF/PBS or DMSO/PBS should be good solvent to dissolve lipases. In some cases tert-butanol was used as a cosolvent with buffer to enhance the activity of lipase.

Hoping this will be helpful,

Rafik

Dear Ajay,

 Mixtures of hydrophilic organic solvent and PBS buffer such as glycerol/PBS,  ethanol/PBS, DMF/PBS or DMSO/PBS should be good solvent to dissolve lipases. In some cases tert-butanol was used as a cosolvent with buffer to enhance the activity of lipase.

Hoping this will be helpful,

Rafik

Dear Dr. Adam, thank you for your help. When I tried last time, I vortex mixed for 15minutes and in the other I just mixed using magnetic stirrer for 3 hours and both created foam. Is that means denaturation of the lipase? Becuase my experiment was also a failure,

Dear Dr. Rafik, thanks for your suggestion. Doesn't the organic solvents may change the nature of lipase?

Hi Ajay, My experience with other lyophilized proteins, not lipase, is that they dissolve within seconds upon gentle vortexing, or just inverting the tube several times. I would never consider vortexing for 15 minutes or magnetic stirring for 3 hours, as that could easily denature enzymes, causing them to lose activity. I suggest you follow up on Rafik's suggestion and look for other solutions with which to make the stock solution. First, check the literature to see what other people have done.

Hello Ajay

Think the undissolved parts of your enzyme preparation result in the structure of the porcine pancreas. Usuallay it is a crudely purified quality, like an Aceton-powder, so it contains hydrophobic molecules from the original tissue that will not dissolve in water. you can add co-solvents, but in your case the foam might be a mycellar effect of hydrophopbic pancreas by-molecules

I would think that the lipase molecules can be dissolved in aqueous buffers.

Usually, Lipases are really stable, that's why you can easyly formulate them in long therm stability in washing powders, togehter with proteases. Im princibable there should be no denaturation of lipase enzymes under lab stirring conditions at room temeperature, you need much higher energies to really denaturate molecules (high temperature, ultrasonic, high energy mixing), but a deactivation can result also from other enzyme impurities, like proteolytic enzymes in your preparation.

If the inhomgenous structure or the foam is disturbing you, you can try to centrifuge and filter the aquoeus preparation before start, if you remove the surface active and mycellar particals, your active Lipase should be located in the clear solution. Undissolved solid lipase might not give any activity to your reaction at all (in case of an aqueous hydrolysis).

Good Luck

Hi Ajay, I bought lipase from porcine pancreas(sigma, L3126) and had the same problem. The lipase could not dissolve in PBS( 5mM) even with 0.1mg/ml and stirred in 37°C. Have you found a way to dissolve it now? I hope to get your help.

Hello tan please tro to dissolve in 1x PBS at pH 7.0 or 8.0. With gentle mixing

Hello,

I also had problems to dissolve the L3126 porcine pancreatic lipase i 1mM tris-HCl pH 7.4, but I managed to prepare a solution using 1 M tris-HCL pH8 (100mg in 2.5ml), gently mixing and waiting for about half en hour.

Please I am trying tO use Lipase from candida rugosa and at higher concentration when I stir, it foams. Does it means the enzymes have been denatured?

Hello Evelyn

No, your enzyme is not denatured, but it might be difficult to get read of the foam. Protein in solution easily foam when stir. So to avoid foam don't stir, but flip the tube gently upside down and wait. it was how I managed to dissolve the porcine pancreas lipase. Good luck!

Actually, foaming should be avoided as much as possible, because it can result in loss of protein activity.

Thank you Sylvie Bolla and Adam B Shapiro . I want to do a release study with Lipase on PCL. Please how do I get to dissolve the Lipase in PBS 7.4 sucessfuly without causing loss of protein activity using a high concentration of the enzyme. Or is there any solubility limit for this enzyme (candida rugosa)? thank you.

When i produce lipase from bacteria and precipitate it by acetone... How can l dissolve it?