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Author: Subject: Action of enzymes on insoluble substances
Baphomet
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[*] posted on 15-5-2011 at 00:10
Action of enzymes on insoluble substances


I have a substance (daidzein) that is insoluble in water and only poorly soluble in organic solvents. I want to use enzymes to convert it to something else, but I'm worried that the nature of the substance will prevent enzymatic action. If it is insoluble then the surface would consist of a 'wall' of molecules, and the molecules would have trouble getting to the active site on the enzyme.

Is there any way around this problem? I'm thinking of glycating the daidzein with a sugar to form glycosides (which have fair water-solubility), maybe using a type of Maillard reaction. It is feasible?




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Baphomet
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[*] posted on 15-5-2011 at 03:02


It looks like it is possible in theory, but it's highly dependent on the substrate and enzymes involved:

http://www.poly.edu/grossbiocat/_doc/publication/Macromolecu...
http://www.nfri.affrc.go.jp/guidance/kankobutu/pdf/kanko_kik...

Sometimes the enzyme may even be adsorbed onto insoluble substances.. which would not be desirable at all.

I should note that the glycation idea would help because even though the glycoside cannot be acted upon by the original enzyme, there are other enzymes that easily convert glycosides back to their corresponding aglycons & sugars.




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[*] posted on 16-5-2011 at 03:26


According to literature, daidzein is soluble in dilute NaOH, and I've confirmed this with a small experiment.

Upon acidification, the daidzein precipitates again, but it now seems less hydrophobic (the material remains suspended in the solution to some extent instead of dropping to the bottom immediately). So maybe the particles are smaller - but still not nanoscale.

It might even be possible to use the enzyme directly on the NaOH-solvated material. Hopefully it will not be denatured by the high pH.




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