fermentation/enzymatic conversion of ibotenic acid to muscimol / amanita muscaria / pantherina

[Prima Materia]

Greetings!

The patent on conversion of ibotenic acid to muscimol suggests best result is achieved with treating the mushroom water extract (tea) with GAD (glutamate decarboxylase) enzymes. It further explains that many of the commonly used Lactobacillus species (such as L. plantarum, L. paracasei, L. lactis, L. brevei, L. delbrueckii, ...) actually produce these enzymes in fermentation.

So the question is --> Did any member of the given forum try to ferment their Amanita muscaria/pantherina tea with LB ? If yes, what was the result achieved, what was the experience like ? Do you think judging by effects that there was a good conversion rate of IBO to MUS ?

Also --> I will conduct the experiment myself. I still have some dry and powdered A. muscaria caps. I'll make some tea with the powder, filter and decant, add table sugar (sucrose) to it as food for the LB and then add a culture to start the fermentation. I'll take the LB from sauerkraut. According to a quick google search sauerkraut should contain the correct LB species to make a good conversion. I intend to ferment at room temperature for a few days and observe the process.

If there are any ideas you would like to contribute or if I had left out anything important, please remind me so I can add or correct.
If any member would like to join in with the experiment, it would be very interesting to compare the results and have a little discussion in the forum.

In yet another alternative embodiment, the filtrate is combined with one or more Lactobacillus bacteria Such as L. plantarum, L. paracasei, L. lactis, L. brevei, L. delbrueckii, or any other fermenting bacteria containing glutamate decarboxylase, or a Substance containing glutamate decarboxylase, Such as rice bran. Thereafter, according to certain embodiments, the filtrate is optionally fermented with the bacteria. According to certain embodiments, the fermented product is
filtered and clarified with or without pasteurization. According to at least one embodiment, the fermented product is filtered and clarified through cotton or other filtration material, and/or filtered through an activated carbon filter.

According to certain embodiments, the filtrate is combined with one or more Lactobacillus bacteria Such as L. plantarum, L. paracasei, L. lactis, L. brevei, L. delbrueckii, or any other bacteria known to contain glutamate decarboxylase (“GAD'). Thereafter, the filtrate and approximately 150,000 colony forming units (CFU’s) of the bacteria per ounce of filtrate are optionally adjusted to a pH of 3.8-5.5 and incubated at a temperature of approximately 98°-155° F.

According to certain embodiments, approximately 0.4 g of CaCO or CaCl per 64 ounces of filtrate is added, along with a prescribed amount of P-5-P as a cofactor (typically 10 mg), and approximately 4.5 teaspoons of table Sugar. Initial pH of the combination of the filtrate and bacteria is approximately 6, and typically drops rapidly within 12 to 24 hours of fermentation to just under a pH of 4. After approximately 3 days of fermentation, the product is filtered, refrigerated, and clarified. The fermented, filtered product is thereafter available for use.

Bioassays of the resultant product show acceptable taste, mouthfeel, and appearance, and may be mixed with fruit juice. The resultant product did not display the undesirable effects noted in fresh A. muscaria tissue.

 

[Prima Materia]

Let me post the notes of how my experiment was/is going.


--> I got sauerkraut today and will start with the process today.

--> It started to ferment nicely at 24 hours from adding the culture from sauerkraut.

--> I'm not very experienced in LAB fermentations. However the fermentation looks very much like yeast fermentation, which I have more experience with. After it will finish I'll check the pH value - hopefully it will be about pH 3.

--> In the fermentation going on there is yeast present as well. Next time I do this I'll have to make everything cleaner so the yeast from my kitchen does not end up in the AM tea. However it will still be very interesting to check the final pH and do a bioassay. The fermentation is still going strong. Will keep you updated.

--> I've also read on the web about yeast also producing the GAD enzmyes. I think the yeast fermented stuff will be less palatable. Hopefully not too bad.

--> It's almost 72 hours from when the fermentation started. The pH strips show it's now at pH 4. I am trying out a small dose of 0,75 grams worth of liquid which is exactly one 30 ml shot. The aroma is not bad at all. Quite strong, mushroomy, slighty acidic and some residual sugar remaning. Maybe a slight hint of alcohol.

--> The effect is not anything worth mentioning. Maybe a slight reduction of anxiety. I had a quite full stomach. I will let thing ferment until it stops. I felt judging by the flavors present that it's not ready yet.

--> It think I'll let it ferment for two days more and check the pH and taste again. My gut feeling also tells I should get more acidity. The residual sugar also show that fermentation is not mature yet.

--> Almost seven days past since the fermentation started and it seems the microorganisms more or less ate all the sugars available. The stuff is now in the fridge. I will soon decant it and filter it if needed and also check the pH again.

--> After 8 days of fermentation I've measured the pH value again - it dropped a little more, now it's at aprox. pH 3,5 - also the perceived sourness increased a bit. Regarding the taste -> It did develop even more and I'm very happy with it. It's very fruity, reminding me of pears and apples. Very interesting stuff. Interestingly the dried and cured A. muscaria also reminded me of fruit. One shot of 30 ml will give a slight reduction in anxiety, very smooth almost non perceptible. Next phase for me will be to bioassay 100 ml of this brew. So far I'm very happy with the result.

--> The recipe: Make a water extract (tea) with 25 grams of Amanita muscaria (fly agaric) mushroom powder. Add 1L of water and a pinch of citric acid and 25 g of AM powder. Aim for a pH of about 4. Don't boil the water but only cook below boiling point and stir well for about 45 min or 60 min. Add 50 g of white table sugar into the mix at least 15 min before the end of the cooking. Pour the tea from your pot into your preferably glass container. Wait until the powder settles on the bottom of your container (best is to wait overnight) - then decant the liquid into another glass container. Then add a few pieces of sauerkraut into the now room temperature tea and mix a little but by turning the container up and down. The volume of your tea should be 1L - this is important for dosing. Make a seal from a piece of aluminium foil - with that you will enable the CO2 from fermentation to escape the vessel and thus prevent a potential explosion. Now wait until the fermentation starts and leave it at room temperature. It should start in about 24 hours after adding the sauerkraut. Wait until the fermentation ends - this can take up to a week or more. After that put the ferment in the fridge. You can also filter everything to get rid of sauerkraut pieces. Now you know - for dosing with this recipe --> for every 100 ml of this brew you have 2,5 g of AM powder. I will continue my research with AM fermentation and keep you updated. If anyone is willing to repeat this experiment and share results I would be very happy.

 

[AliRadicali]

I think this is a fascinating experiment, and I thank you for sharing it, but for A. Pantherina specifically, I don't think this process is worth the effort if the data from this paper are representative: http://citeseerx.ist.psu.edu/viewdoc/download?doi=10.1.1.688.466&rep=rep1&type=pdf
According to the paper, they found 2-3 times more muscimol than ibotenic acid in the (dried) pantherina caps, whereas in (fresh) A. Muscaria you get up to ten times more ibotenic acid than muscimol. As such the main concern should be properly dosing these mushrooms, not so much converting IBO into MUS, although cooking at the right temp and adding lemon to the tea is always a good idea.
Still, I love the experiment and I might try it with some of my muscaria samples.

 

[Prima Materia]

I think this is a fascinating experiment, and I thank you for sharing it, but for A. Pantherina specifically, I don't think this process is worth the effort if the data from this paper are representative: http://citeseerx.ist.psu.edu/viewdoc/download?doi=10.1.1.688.466&rep=rep1&type=pdf
According to the paper, they found 2-3 times more muscimol than ibotenic acid in the (dried) pantherina caps, whereas in (fresh) A. Muscaria you get up to ten times more ibotenic acid than muscimol. As such the main concern should be properly dosing these mushrooms, not so much converting IBO into MUS, although cooking at the right temp and adding lemon to the tea is always a good idea.
Still, I love the experiment and I might try it with some of my muscaria samples.

AliRadicali, thank you for uploading the study. I'll have a look at it. It seems like great info.

I would have to get the sample tested to really be sure about the IBO to MUS conversion.

So far I have not delved yet into bigger doses to have a bioassay and see the potential unwanted side effects of possible ibotenic acid in the fermented sample.

Interesting info about the pantherinas as well.
I have yet to try those. I still have some dried caps. I have found when curing the dried caps that the smell of dry A. muscaria has a much richer aroma, very fruity, floral like. I have also observed this with this fermentation experiment - again getting those fruity aromas. The panther caps had a more dry or chemical aroma to it when curing - thus suggesting that they have a much different chemical composition. I have a feeling that the various other compounds in A. muscaria contribute very much to it's healing potential. Uploaded is a study explaining into detail about the other compounds as well, and not only IBO and MUS. Also going by gut feeling I would choose A. muscaria over pantherina for trying to get the health benefits seeking. However, I am interested to know about the other compounds in pantheria as well.

I have another fermentation experiment going that I'll be reporting about as well.

Please do write or upload, if you find any relevant info on the topic.

 

[AliRadicali]

I concur, Muscaria has the more pleasant aroma, dried Pantherina has a strong, sour smell to it, which is present but not dominant in the muscaria.