Has anyone tried to perform separation using cellulose digestion? Using cellulase, ligase, etc., it should be possible to completely dissolve the cellulose that composes the cell structure.
The breakdown products from the cellulose are simple sugars, easily removed.
With the cellulose gone, the cell contents are released. I would assume that desirable compounds could then be recovered from the slurry.
Cellulose digestion would be a pre-processing step, followed by compound separation, via different solvents.
My thinking is that 100% extraction efficiency should be the goal.
By dissolving the cell membranes, everything will be released into the sugary liquid slurry.
And by everything, I mean exactly that. At the point where the cell walls are digested, you have a liquid that contains the entire essence of the plant.
Discussion about compounds of benefit, beyond THC, is common. If you wash your biomass, with the cellulose intact, it is something like spraying a muddy fence with a hose. You can clean off all the mud on this side, but the fence is in the way, so mud lingers.
Remove all the cellulose first, then wash away the sugars, leaving everything else. Use washes to get what you want.
I think it’s an interesting idea, are those enzymes the same kind used for biofuels?
Would the enzyme treatment be done in water or alcohol? Maybe even you could isolate the sugar and make rock candy.
feed it to yeast and make extraction solvent or fuel.
the strategy of dissolving the cell walls works best if what you’re after is INSIDE those cell walls. If you’ve got a use/market for all (or at least some of) the other components, it might be a win.
dissolving fresh biomass in a vat with enzymes, then centrifuging to get the water solubles separate from the non-solubles, and then extracting the hydrophobic fraction with your solvent of choice might get you there…but you’d lose the water soluble terps.
I think his point was that by degrading cellulose it would not be able to adsorb/absorb/retain his product in theory, am I right? So is our cellulosic biomass causing cannabinoid retention (acting like a paper or cotton filter?). I hadn’t thought about it until now. I wonder what kind of yeild difference we’d be looking at here
so long as you want the cellular contents.
my contention is that we don’t.
works great if you’re after say lycopene.
high energy sonication also isomerizes cannabinoids
(which was a win with lycopene…)
http://www.plandaibiotech.com/ was on this for a bit. they seem to have vanished. they didn’t believe me 5 years ago when I told them that screaming at my girls was a bad idea…but it clearly wasn’t a good one…
cellulose makes awesome filters. isolating hemp cellulose and selling it to folks for filtration media makes a lot of sense. but I personally would go at that post cannabinoids
Not certain on “100%”, and there are certainly trichomes that are not up on pedestals to make them easy targets.
In intact trichomes, the cannabinoids are dissolved in terpenes (terpenoids, terpenols, etc)…they’re not soluble in water, so the plant only has so many places to stash them. The solvent(s) are also fairly specialized secondary metabolites.
Are all the terps in the trichomes?
No. Isoprene is made elsewhere, and it would be ludicrous to suggest that building block was used nowhere else.
Dissolving the cell walls and blowing every compartment open to get at the cannabinoids feels more like bulldozing the house to get at the beer sitting in the fridge on the front porch.
Don’t forget about the phospholipid bi-layer and the work required to get through extracellular matrix proteins. I think the pre-processing step might need a pre-processing step.
Enzymes for most things can now be selected. Once we really get a handle on protein structure, we should be able to design them deliberately. Once we really get into synthetic biology, and get to play with hundreds of different amino acids rather than a couple of dozen, the reactions that can be enzymatically catalyzed should be essentially infinite.
I have been looking at a variety of sources and it appears that cellulase is not extremely expensive. It should be noted that enzymes act as catalysts, and are not consumed by the reactions that they catalyze…a little goes a long way.
Cellulase is an enzyme that breaks cellulose down into glucose. So after a passive treatment (stuff sits in a vat), your biomass is dissolved and you end up with sugar water that has all of the water insoluble stuff either floating on it, suspended in it, or precipitated out.
Plant elements (non-polar compounds) that are not dissolved in the water phase, of the enzyme bath, can be separated through filtration. It may be possible to isolate desirable compounds, using just this filtration.
The water phase would not be wasted either. Separate out the sugar water and use it to make ethanol, if you want to, just throw in some brewer’s yeast.
I have suggested enzymes as a pre-treatment, but it could just as easily be used to post-treat whatever biomass remains after standard extraction. No more spent biomass waste disposal, plus you reclaim that final bit that would have been thrown away.
If you don’t want to make alcohol, you could also process pure glucose powder. And in the case that you just don’t want to be bothered at all, it is sugar water, nothing hazardous at all. Disposal should be simple, unlike biomass that can contain residual solvents.
if the “additional” cannabinoids are essentially a side effect of turning a waste stream into an income stream, the strategy makes way more sense to me.
I understand the concept of wanting to use the whole plant.
I just have a hard time with adding a whole bunch of stuff you don’t want to the bit you do want before you try and purify it.
The general concept of using cellulose to make alcohol is not normally a preferred commercial method, but as a secondary income stream, it makes a lot of sense.
For the producer that is already using ethanol in their SOP, it makes even more sense. I suspect that post-treatment of spent biomass containing residual ETOH (after a wash) would not be problematic. In fact, it has been found that the cellulases work better if yeast is around too. The sugars produced are quickly converted to ethanol, and don’t hinder the enzymes.
