Ion-exchange Chromatography with Enzymes

Rod sciteach at droog.mbt.washington.edu
Tue Jul 18 12:52:18 EST 1995


We're working in a summer program for Middle School Teachers, trying to 
develop some labs to use with kids in the 11-14 age range.  One method 
we're trying is using an ion exchange column to separate two enzymes in 
solution and show their presence in collected fractions by showing their 
activity.  We've been using alkaline phospatase (bovine intestinal 
mucosa) as a sample, but aren't having much luck in getting it to bind 
to the matrix-- (Bio-Rex 70 - cation) and DE-52 (anion) are two that we 
have on hand.  We've been testing for AP with pNPP at this point. 

We've tried a ph range from 4-8.  Is there a way to find the optimum pH 
for a particular enzyme/resin combination?

 Ideally we'd like to find two enzymes from some natural source--like 
banana or potato extract, or saliva, or beef liver--maybe 
peroxidase,amylase, catalase or AP that would would show up with a color 
indicator that is relatively safe to use in the classroom.   We've   
been using "fast red" as and indicator for AP in some labs, but any 
enzyme/source/indicator combinations that you know might help us.  Or 
where else would you send us to ask?

Thanks in advance for your help,

Rod Margason
Integrated Science Partners 



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