Hi, Research Community
I am working on a two-step enzymatic reaction, wherein 1st step requires Mn2+ while 2nd step needs Mg2+ supplementation for optimum cleavage of substrates. The presence of additional divalent cations in either of the steps triggers non-specific activity.
Any suggestion/recommendation on which chelator can be used to selectively chelate the Mn2+ post-1st step before I add Mg2+ and perform the 2nd-step reaction?
Thanks in advance.
To clarify: is the system you are testing using a single enzyme in which Mn is required in the first step, followed by Mg in the second step but the same enzyme, or are there two enzymes, one with Mn the other with Mg?
And then you want a chelator strong enough to remove Mn but not so strong or general to allow Mg to take its place in the enzyme?
My knowledge of chelators is limited to EDTA, HEDTA, EDDHA etc, but short of testing each one (at specific combinations concentrations of Mn Mg and chelate) and pH, you might be very lucky to find a specific chelator for Mn vs Mg. Perhaps some of the resins used in chromatography/water purification might work, but you may still need to do a lot of testing to find the right combination.
Hi Dr. Matthew, thank you for your suggestions. I am working on it to find an optimal combination.
To answer your query: Yes, its a single enzyme catalyzing both the reactions.
The chelator which can chelate magnesium will also do more or less the same effect on mangnese too. For the separation of these two metals you have to seek aid of definite ion exchange resins and also subjected to definite pH.
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