Lipase extraction

[cristianammg]

Hello everyone (This is my first post here)

 

I'm a PhD student (More Info) and I need a method to extract the produced lipase from an effluent. This lipase is from Y. lipolytica or C. cylindracea. I found lots of methods on the Internet and articles/papers but I want something really simple/rudimentary that doesn’t significantly modify the medium were lipase was produced.

 

 

Could you please help me?

 

 

Thank you all…

[CaptainPanic]

Lipases are enzymes - proteins essentially.

 

As far as I know, a change in pH can change the solubility of proteins a lot. If they are soluble, you might convert it into insoluble, and use a filter.

 

Alternatively, you might try a real extraction, with a solvent (any water-insoluble solvent might work). But in a biological medium, any solvent will extract a lot more than just your enzyme... and it might kill your microorganisms.

 

Can't you somehow immobilize the enzymes while they're still in your medium? I know there are tricks to immobilize enzymes, but I am not familiar with the techniques.

 

p.s. (1) I'm a chemical engineer - therefore only half-qualified to give you advise.

p.s. (2) have a look in our other sub-forums too. We also have a Biochemistry and Molecular Biology forum that might interest you.

[cristianammg]

Lipases are enzymes - proteins essentially.

 

As far as I know, a change in pH can change the solubility of proteins a lot. If they are soluble, you might convert it into insoluble, and use a filter.

 

Alternatively, you might try a real extraction, with a solvent (any water-insoluble solvent might work). But in a biological medium, any solvent will extract a lot more than just your enzyme... and it might kill your microorganisms.

 

Can't you somehow immobilize the enzymes while they're still in your medium? I know there are tricks to immobilize enzymes, but I am not familiar with the techniques.

 

p.s. (1) I'm a chemical engineer - therefore only half-qualified to give you advise.

p.s. (2) have a look in our other sub-forums too. We also have a Biochemistry and Molecular Biology forum that might interest you.

 

Ok... thank you very much... I'll search in other areas and I'll take your tips in consideration

[hermanntrude]

two possibilities you should always be aware of when doing a PhD:

 

1) no-one has ever done it before. you may have to painstakingly and slowly develop the method over several years

2) it cannot be done. You may have to spend years and years painstakingly developing the best possible method only to discover it's impossible.

[Psycho]

I think the best option would be to use various gel chromatography, to purify the protein based of size, charge or any unique feature of the protein. If worst comes to worst you could always raises antibodies against it and attach them to beads within the column, hardly an economical way of achieving purification though.