Enzyme Assisted Extraction (EAE)

Carbon Chemistry has pioneered the use of both acid and enzymatic degumming, simultaneously distributing materials for using those methods at the top level. This post is meant to announce a quite outstanding alternative use of our enzyme product, Enzyme Assisted Extraction…a new type 6 extraction method that needs further field trials to verify.

“Demand for new and novel natural compounds has intensified the development of plant-derived compounds known as bioactives that either promote health or are toxic when ingested. Enhanced release of these bioactives from plant cells by cell disruption and extraction through the cell wall can be optimized using enzyme preparations either alone or in mixtures. However, the biotechnological application of enzymes is not currently exploited to its maximum potential within the food industry. Here, we discuss the use of environmentally friendly enzyme-assisted extraction of bioactive compounds from plant sources, particularly for food and nutraceutical purposes. In particular, we discuss an enzyme-assisted extraction of stevioside from Stevia rebaudiana, as an example of a process of potential value to the food industry.”

So what does that mean and why the F do you care?

Enzymatic degumming attacks lipids and hydrolyzes specific portions to break a complex compound into its constituent compounds, which makes them easier for us to remove.

Have you ever wondered what the wall of a trichome is made of? Its made of lipid compounds that the enzyme degrades. What happens when lipophillic compounds are released in water? They float to the surface. The thesis of this method is ‘Liberating the compounds in trichomes without a carrier solvent interacting with the yield’.

Ok, so here is the money shot: You place cannabis trim/flower in water, adjust to proper pH and temp, add and agitate enzyme gently, press the biomass to the bottom like a French Press, decant the organic phase off the top of the water, dehydrate into cannabinoid/terpene/etc product. The primary post processing requirement is removing fatty acids form the organic phase.

Water extraction with a supercharger.

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Sounds legit.

You got a link to that paper that isin’t behind a paywall?

or u can extract with countercurrent compressed co2 and avoid the organic solvent.
i cant find a real copy but
Publication : USDA ARS

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I have found that Natures Miracle Dog Urine Remover will remove the stickiness that accumulates on surfaces like counter tops from this extract and fiddling with it like I do. Little drips and drops makes things sticky.

You have to let the urine remover sit for a while and I dose it a few times but then extract residue can be wiped up mostly with very little or any solvent except soap and water after several hours. The stuff is pet and children safe.

It is millions of live bacteria in the stuff that neutralize the dog urine and it seems to work great around my lab areas where I clean up to get that stickiness from the extract labs off of everything. The bacteria eat the stuff by secreting enzyme and when the food is gone then they go dormant. When more food shows up they start munching again so it tends to also keep the counter from staying sticky long. First soak has to be long but after it sure stays brighter with time.

That is 100% enzyme action.

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If you load the DOI number from the title of the paper into www.Sci-Hub.hk you can read the entire paper.

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So that would be:

  1. Submerge material in warm water (for enzyme optimization)
  2. Add enzyme and agitate gently.
  3. Compress biomass to yield aqueous and organic layers.
  4. Decant oil/water from extraction vessel.
  5. Countercurrent CO2 to separate the organic phase?

We will be helping people test this method with our enzymes over the next month or two until we have some results to verify the usefulness of this method.

The primary concerns are removing fatty acids from the oil yield, and achieving optimal extraction efficiency.

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I can haz?

I don’t have access to the tek for Countercurrent CO2, but I’ve got kief and a press pot :wink:

I havent read anything with countercurrent co2 for cannabis so i am very interested in that test. what kind of countercurrent column ru going to use?

Tanks. sci-hub.hk no good. sci-hub.tw does the trick.

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We are still developing the method from a theoretical to a practical. The primary data point we need before moving that direction would be extraction efficiency prior to separation, followed by analytics of the yield.

Equipment wise, we seek to observe what options are available for an industrial french press and a decanting method(centrifuge?) that can be performed under a nitrogen backfill. Shooting for a low oxidation, ultra high quality, non-volatile production.

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what about the equipment for the actual countercurrent co2 part? the liquid- liquid centrifuges sound lik a good choice for seperation

This work is super interesting to me after all the research ive been doing recently is related

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From what I gather about co2 liquid expansion, a pressure vessel with a collection method at the top would be ideal. Expanding the liquid level to the collector for partitioning while improving penetration is a desirable system design.

This method uses water as the co-solvent so I wonder if there would be any side reactions.

depends on the pressure u go at. if ur talking about “liquid” co2 not “supercritical” co2 cause both have been used in this process.
the pressure determines the ph do to the in situ formation of carbonic acid
Dense CO2 as a Solute, Co-Solute or Co-Solvent.pdf (179.9 KB)
Dynamic pH determination at high pressure of aqueous additive mixtures in contact with dense CO2.pdf (639.6 KB)
Reversible in Situ Catalyst Formation.pdf (366.9 KB)
Title Fractionation technologies for liquid mixtures using dense carbon dioxide.pdf (702.8 KB)
Deodorization and Deacidification of Edible Oils with Dense Carbon Dioxide .pdf (619.9 KB)
Countercurrent fractionation of aqueous apple aroma constituents using supercritical carbon dioxide.pdf (901.1 KB)
Packed column supercritical fluid chromatography of hydrophilic analytes via water-rich modifiers.pdf (812.3 KB)
The Use of Dense Carbon Dioxide Extraction and Fractionation to Recover and Refine Natural Food Ingredients from Food Processing Wastes.pdf (5.7 MB)

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Can’t we use more water to wash/float the organic layer away?

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I’m assuming it will contain more then just cannabinoids that way but if that ends in a good end product then that would awesome. plus without an extraction step there might still be enzymes in it at the end

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That’s the direction I’m leaning towards for R&D. The ‘tilt and fill’ method :wink:

You can deactivate the enzyme at 60C and use a filtration element in the press or pour through one.

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Via counter current distribution, manys thanks to good old Lyman

Has anyone followed up on this? I would like to collaborate on this and possibly get the research funded. Let me know if anyone is interested.

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I followed the papers and attempted to order one of the enzymes but the chemical company told me the product was a controlled substance and refused to sell it to me.