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 precursor) 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 technique.
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 fruit bodies grew dwarfed.
Bio-assay showed that they are at least 3 times the usual potency.
PRESERVING THE PSILOCIN - Use the cool desiccation technique of the
PF TEK. Dry the shrooms in a refrigerator 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.
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 subs-tropical 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
precursor than tryptophan (5-7). It was found that not less than 22.4%
of the psilocybin formed was derived from the labeled precursor
tryptamine (5). The level of psilocin was generally zero in the
mycelial tissue from these experiments (5-7).
In the present paper, the bio-transformation of fed tryptamine in
fruiting mycelia of Psilocybe 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 Psilocybe 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 Psilocybe 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 bluing 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 Psilocybe 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 Psilocybe 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 Psilocybe 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 bio-synthesis of
psilocybin from psilocin in the media.
P.cubensis also failed to produce detectable amounts of baeocystin
under these culture conditions.
Acknowledgments
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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