Enzymes are typically large molecules with molecular weights >10,000. If the complex hydrocarbons are much smaller, you may be able to remove the enzymes by passing the seawater through an ultrafiltration membrane with a 10,000 molecular weight cutoff. You would have to validate that the hydrocarbons did not stick to the membrane.

Another approach would be to add an ion exchange resin such as Q-Sepharose to the seawater to adsorb the proteins on the basis of charge. Hydrocarbons, being uncharged, would not stick to the resin. Once again, you would have to validate that the hydrocarbons did not stick to the resin.

Another idea is to destroy the enzymes by adding another enzyme, a protease, to chop the enzymes up into smaller pieces. The pieces would remain in the sample, but would no longer have the undesired activity. The protease would also chop itself up after a while.

Still another idea is to heat the sample to 70 degrees C or so to denature the enzymes. Unless they are from thermophilic bacteria or are unusually stable, most enzymes would be inactivated by heating. You would have to be careful not to evaporate the volatile hydrocarbons.

Hello,

Depending on the optimal pH and temperature of your enzyme, you can change one of these variables to inactivate the enzyme. For example, adding 100-200 mM Na2CO3 (a weak base) will turn the pH of the solution around to11-12. This should not degrade carbohydrates, but inactivate most enzymes. Rather, you could add citric acid, which at low concentrations also doesn't break down carbohydrates. Also, a mild thermal shock, i.e. 70-85 ° C for 10 min, should not affect carbohydrates.

All the best

Carlos

You could try to use immobilized inhibitors or immobilized high affinity ligands (on beads). For intance lectins and/or large spectrum protease inhibitors. If you are in luck you will clear your solution of the toxic components by soaking with the bead and centrifuge.

Good luck

Philippe

Go heat inactivation both protease and amylase enzymes.

Cheers all. These are useful, although we are limited to approaches that are less likely to destroy or affect the hydrocarbons we are interested in. This rules out heating, and to some extent high temperatures, although what a high temperature is in this context is open to question. Adam's (and Philippe's) suggestions seem like the best solutions so far, in terms of practicality and probably cost as well. I will steer the research in that direction, and make good use of positive and negative controls (as always) to see if we lose anything in the process.