Ion-exchange Chromatography with Enzymes
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,
Integrated Science Partners
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