Psilocybin production increase

[boingo]

greetings!
 have any bees successfully increased the psilocybin content of their homegrown p. cubensis (rice cake method)?

one method i saw was mentioned by psylocybe fanatacus, who used tryptamine in the bell-jar water.  they claim 3x the potency of
the unenhanced fungi. 

for something that looks so easy to do, i'm surprised i haven't heard more about it.

bees?
peace,
boingo

[DroYd]

No, it allways add enough in there, for me.

Nothing like a freshly cultivated Stropharia Cubensis. Awesome, first thing in the morning, and I'm good all day.

Oh, but, you are talking about a differant sub species. I think I read once, that differant species, had differant amounts, of Psilocybin and Psilcyn, in them.

[MethGn0me]

there are other substituted precursors you can give the plants that will yield very good chemicals
i've heard of giving some hallucinogenetic plant(i dunno which one) dopamine and it somehow converts it into mescaline.
there was a thread at lycaeum called biosynthesis via substituted precursors
read it

[psyloxy]

those hallucinogenic plants are peyote/san pedro cacti

[jonah3]

I read something on the shroomery about using Desmanthus Illinoesis bark or root bark. I'll see if I can find it.

[jonah3]

Potency of P. Cubensis: A Successful Dream.
by phase two

This is a fascinating technique just send in by an unidentified shroomer out there. If anyone tries this, please post the results on the message board.

Last night, I dreamed I took part in a successful experiment involving strange plants and lifeforms I did not completely understand . When I awoke, I wrote the following monologue in my dream journal. It is a work of -fiction- from my creative unconscious. I take no responsibility for anyone attempting to live out my strange dreams!

"120 grams of Desmanthus Illinoesis rootbark was broken into small pieces by hand, and these 'chips' were added to a blender. They were blended on high speed, and shaken, until the bark was reduced to a fine, rusty-colored powder. The blender was left with the lid on for about 10 minutes, allowing the fine particles to completely settle before opening. The contents of the blender were carefully emptied into a metal cooking pot (with a fitting lid). The pot was then partially filled with Denatured Alcohol. This pot was taken outdoors in a well ventilated area, covered, and gently boiled on an electric hotplate for several hours. (DANGER: DO NOT BOIL DENATURED ALCOHOL IN AN UNVENTILATED KITCHEN!). After several hours, the solution was allowed to cool , and then strained through a screen strainer with a coffee filter resting on top of it. The resulting alcohol was deeply rust colored. (A bit of this alcohol solution was 'tested' for possible tryptamine content, by pouring a small amount onto a glass dish and evaporating it. The resulting material was a yellow-tan crystalline substance that looked excitingly like synthetic DMT is described. Scraping this material with a razorblade and smoking the residue showed it to have a -threshold- of psychedelic activity, yet beyond any possible placebo effect, in the opinion of this researcher.)

A 'deep dish' baking pan was filled with approximately 3-4 cups of horticultural vermiculite, of a grade suitable for use as a mycological substrate. The rusty-colored alcohol solution was stirred slowly into the vermiculite carefully, to expose it to the vermiculite as evenly as possibly. This baking pan was put outside, on the hotplate (medium setting) to expedite the evaporation of the alcohol. The vermiculite was stirred approximately every 5 minutes to make sure the alcohol was evaporating and leaving tryptamine deposits on the vermiculite evenly. After approx. . one hour of heating and stirring, the vermiculite in the pan -appeared- to be completely dry, sandy in texture. After CAREFULLY wafting (smelling) the pan from a distance, it was determined that it still smelled like alcohol. (DO NOT SNIFF UP A LUNG FULL OF HOT DENATURED ALCOHOL FUMES!!!) After heating and stirring for 15 more minutes, the vermiculite was carefully determined, by smell and texture, to be dry and completely free of alcohol. It was then taken inside, and put in the freezer to cool.

This 'enhanced' vermiculite was soon used to prepare a mycological substrate, adhering to the PF method. The jars were prepared, sterilized, cooled and inoculated as normal. Mycelium growth occurred at the normal rate. The cakes were cultivated in a styrofoam cooler, with perilite humidification. The fungi grew quite normally. Upon drying and sampling, they were subjectively considerably more potent than p. cubensis grown on normal vermiculite. One freshly dried mushroom, weighing .6g, was all that one experienced explorer could comfortably handle. For information as to how this process may have increased the potency, see page 280 in TIHKAL. These are strong entheogens! Treat them with love and conscious respect, or they may 'punish' you. Please keep them sacred and do not involve those souls who do not genuinely seek to experience and live with the Divine Light!"

[jonah3]

TRYPTAMINE CUBENSIS
entered: Jan 10 1997 by Psylocybe Fanaticus - Seattle
In the following transcription of the science paper and discovery of Dr. Jochen Gartz, he describes adding a 25 millimolar concentration of Tryptamine HCL (a psilocin and psilocybin precurser) to the Cubensis substrate and under lab control conditions, discovered the potentiation of psilocin into never before measured levels in Cubensis fruitbodies of up to 3.3% psilocin which is several times the potency as regular Cubensis.

PF TEK application of the Gartz Tryptamine tecknique.
1/2 pint jar:

1/2 - 2/3 cup of vermiculite + 1/8 cup of brown rice powder and (45 milliliters of water with .16 grams of Tryptamine HCL added)

PF experiment results:

The fungus cultured as usual except that the fruitbodies grew dwarfed. Bioassay showed that they are at least 3 times the usual potency.

PRESERVING THE PSILOCIN - Use the cool desiccation tecknique of the PF TEK. Dry the shrooms in a refridgerator under COLD conditions. Store the dried fungi in a tight container with desiccant in the freezer.

March 6 1997 entry - Tryptamine formula update
The above formula is a bit to much for the pf tek. That is why the shrooms grew dwarfed and some jars failed to fruit. The answer is that the pf substrate is much lighter and thinner than Gartz' substrate. Gartz used cooked brown rice and cow dung. This is heavier, thicker and more nutritious than the pf substrate formula, so therefore, the tryptamine hcl content should be less also. PF has received some very reliable information that 1/2 to 3/4 of the above formula should be used. So instead of .16 grams of tryptamine hcl, use .1 or less grams of tryptamine hcl.
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Planta Medica 55 (1989) page 249 - 250 Jochen Gartz
BIOTRANSFORMATION OF TRYPTAMINE IN FRUITING MYCELIA OF PSILOCYBE CUBENSIS.

Jochen Gartz
Institute of Biotechnology, Academy of Sciences of the GDR,
Permoserstrasse 15, GDR-7050 Leipzig, German Democratic Republic

Received: March 13, 1988

ABSTRACT

Mycelial cultures of Psilocybe Cubensis, with the ability to form psilocybin and psilocin de-novo, also hydroxylated and methylated fed tryptamine to give psilocin in up to 3.3% dry mass of the obtained fruit bodies. By using HPLC and TLC, it was found that these mushrooms contain only a small amount of psilocybin (0.01-0.2% dry mass). The values of psilocin are the highest described in any mushrooms.

INTRODUCTION

Psilocybe Cubensis (Earle) Sing, is a substropical mushroom and contains the indole alkaloid psilocybin and only small amounts of its dephosphorylated counterpart psilocin (1-4). Variations in these metabolites have been well demonstrated by investigations of fruit bodies cultivated under controlled conditions of a rye-grain medium (2) and rice substratum (3), respectively.

The study of psilocybin biosynthesis in submerged culture of P. Cubensis showed that radioactive tryptamine functioned as a better precurser than tryptophan (5-7). It was found that not less than 22.4% of the psilocybin formed was derived from the labelled presursor tryptamine (5). The level of psilocin was generally zero in the mycelial tissue from these experiments (5-7).

In the present paper, the biotransformation of fed tryptamine in fruiting mycelia of P. Cubensis is described.

MATERIALS and METHODS
Cultivation of Psilocybe Cubensis

A dried cow dung/rice-grain mixture (2:1) with twice the amount of water was used to obtain fast fructifications without casing of a strain (3) of P. Cubensis. A 25 mM concentration of tryptamine (as hydrochloride) was added to this medium. Cultivations without the addition of tryptamine were also tested. The methods of cultivations were described in (3).

The first sporocarps were produced by cultures of P. Cubensis in 3 to 4 weeks. The cultures continued to produce mushrooms in five flushes. Each flush was harvested as soon as the sporocarps were mature. The mushrooms were immediately freeze-dried, sealed in plastic, and stored at -10 degrees C until analysis.

EXTRACTION and ANALYSIS The extraction procedure and the analysis of the indole alkaloids by using HPLC and TLC were described in the previous papers (3,8-10). The presence or absence of tryptamine was demonstrated by TLC as described by Stijve et al. (11).

RESULTS and DISCUSSION

The cow dung-rice mixture actually produced the first flush of mushrooms earlier than the cultivations on ry (with casing) (2) and rice (3), respectively. They yielded an average of 3 g dry mass per 10 g substratum.

Under the same culture conditions, the fructification times, the yields, and sizes of the mushrooms as well as the blueing feature (3) were equal when the growth media also contained high concentrations of tryptamine. Initial experiments without the addition of tryptamine were performed to determine the content of psilocybin and psilocin in comparison with experiments using other culture conditions and/or media (2,3).

The levels of psilocybin and psilocin varied from one flush to the next, but generally were much the same as those in the other experiments (2,3) (table 1). Consistently low levels of psilocin were found in the mushrooms without the addition of tryptamine to the substratum. Additionally, psilocin generally was absent in the first flush as was also observed in earlier investigations (2,3). Table 1 shows that the fed tryptamine gives high values of psilocin in each flush from the cultures.

Table 1 Variation of psilocybin and psilocin levels in P. Cubensis as a function of flush number from the cultivations with (a) and without (b) addition of tryptamine (25 mM concentration).

Flush no. Psilocin Psilocin Psilocybin Psilocybin
 a  b  a  b 
1. 2.1 - 0.01  0.55
2. 3.3 0.01 0.02 0.48
3. 2.8 0.02 0.2 0.51
4. 3.1 0.09 0.07 0.46
5. 2.9 0.15 0.13 0.61

These psilocin levels are uncommonly high (from 2.1 to 3.3%) since values reported for psilocin in dried mushrooms are always below 1% (1-4,12,13).

Inocybe Aeruginasens Babos contains only traces of psilocin but high amounts of the incompletely methylated psilocybin (baeocystin) (9). In contrast to the intitial experiments without an addition of tryptamine, the mushrooms generally contained only small amounts of psilocybin. The tryptamine level was always zero in each mushroom. In this case no tryptamine was additionally found in the methanolic extract of the vegetative mycelia from the substratum.

In a previous report, Gartz (3) was unable to detect baeocystin in P. Cubensis. But Repke et al. (14) reported traces of baeocystin in other strains of P. Cubensis about 10 years ago. They suggested that many non-specific enzyme systems exist in fungi which have the ability to oxidise exogenously added compounds, as well as normal, obligatory intermediates (14).

The results in Table 1 show that the enzyme systems in P. Cubensis have a high hydroxylation and methalation capacity to convert added Tryptamine to psilocin. It is possible that a reduced amount of phosphate in the culture media decreased the biosynthesis of psilocybin from psilocin in the media.

P.Cubensis also failed to produce detectable amounts of baeocystin under these culture conditions.

Acknowledgements

The author thanks the following persons: G. Drewitz, T. Stijve, G.K.Muller, and M. Gey who generously supplied valuable information.

REFERENCES

Heim, R., Hoffman, A. (1958) Compt. Rend. 247,557.
Bigwood, J.. Beug, M.W. (1982) J. Ethnopharm. 5, 287.
Gartz, J. (1987) Beitrage zur Kenntnis der Pilze Mitteleuropas 3, 275.
Badham, E. (1984) J. Ethnopharm. 10, 249
Agurell, S., Blomkvist, S., Catalfomo, P. (1966) Acta Pharm. Suecica 3, 37.
Agurell, S., Nilsson, J.L.G. (1968) Acta Chem. Scand. 22, 1210.
Agurell, S., Nilsson, J.L.G. (1968) Tetrahedron Lett. 1063.
Gartz, J. (1985) Pharmazie 40, 134.
Gartz, J. (1987) Planta Med. 53, 539.
Semerdzieva, M., Wurst, M., Koza, T., Gartz, J. (1986) Planta Med. 52, 83.
Stijve, T., Hischenhuber, C., Ashley, D. (1984) Z. Mykol. 50, 361.
Beug, M.W., Bigwood, J. (1982) J. Ethnopharm. 5, 271.
Ohenoja, E., Jokiranata, J., Makinen, T., Kaikkonen, A., Airaksinen, M.M. (1987) J. Nat. Prod. 50, 741
Repke, D.B., Leslie, D.T., Guzman, G. (1977) Lloydia 40, 566.

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