DD266811A1 - PROCESS FOR OBTAINING INDULSES BY CULTURING AGARICALES AND NUTRIENTS HEREFUER - Google Patents

Abstract

The invention relates to the microbial recovery of indoles. It is applicable for the production of a model substance for experimental, psychiatric research and neurobiological investigations. The invention consists in the use of a new nutrient medium, which consists of natural waste with the addition of water with or without the addition of urea. The non-sterile fermentation of Psilocybe cyanescens takes place at 2 to 32C, the extraction and isolation of the active ingredient in a known manner. The solution according to the invention leads to a largely constant yield of psilocybin with simple fermentation and separation.

Description

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title

Process for obtaining indoles by cultivating agarics and nutrient medium therefor

Field of application of the invention

The invention relates to a method for obtaining Tndolen by artificial cultivation of mushroom mycelia on suitable nutrient media. The substance psilocybin is applicable as a model substance in neurobiological research.

Characterization of the known technical solutions

The indole compound psilocybin occurs in natural fruiting bodies of various fungus species of the agaricales of the genera Psilooybe, Strophariaj conocybe, Inocybe, Pluteus and Panaeolue.

Ee is known that from the cultured mycelia of various species of the drug can be obtained in addition to small amounts of minor alkaloids such as Peilocin and Baeocyetin »

* Thus, a method (DE-OS 1087321) describes the recovery of the compound of mycelium and sclerotia, which have been grown by cultivating fungi of the genera Psilocybe and Stropharia in surface cultures using malt extract in the presence of various mineral salts; The disadvantage of the method is that must be fermented at constant temperatures, which requires additional equipment expenses. Finally, a sterile working is necessary / because the substrate used is also a suitable breeding ground for a variety of faster-growing bacteria and lower fungi H.

This Naohteil hot also a ready proposed method _for that when using

Synthetic nutrient media causes a higher accumulation of indole alkaloids at variable fermentation temperatures in the mycelia ale in the former method. The starting materials in both methods are expensive separation of the mycelia prior to the extraction of biomass is required as an additional process step each time.

Object of the invention

The aim of the invention is a process for obtaining indoles from fungus mycelia, in which using cheap »natural raw materials cultivation is effected under unsterile conditions and at variable temperatures, wherein the additional method step of the previous separation of the mycelia omitted and largely constant yields of psilocybin despite different substrates to be achieved.

Explanation of the essence of the invention

The object of the invention is to develop and use a substrate as a nutrient medium for the culture of the mycelia of psilocybin-producing species. According to the invention, the cultivation of the fungus mycelia is carried out on natural waste materials in the sense of a solid phase fermentation. ¹ The substrates are mold-resistant and are not measurably degraded by bacteria during the usual cultivation times; This makes an unsterile fermentation possible.

As substrates are used

- fallen branches and foliage of Quercus, Titia, Salix, Acer, Aeeculua, Populus, Betula, Carpinus, Alnus, Fagus

- Dropped needles of Picea and Pinus

- Plant dead plants of Urtica in any mixture or as monosubstrate with or without the addition of urea in quantities of 0.01 to 0-3%.

As a cultivated mushroom species Psilocybe oyanescens Wakefield emend Kriegisteiner is used, ¹ Through the use of these mushrooms * is a cultivation at 2 to 32 ⁰ C without any Temoeraturregelung possible.

The procedure is such that the mentioned organic wastes, depending on the moisture content with 10 to 100 % water, based on the solid substrate, are added · ¹ After autoclaving and cooling is at the above temperatures with a suspension of mycotic flakes of Psilocybe cyanescens from a Inoculated by known methods preculture and the culture unoterilically 30 to 50 days left to itself · Thereafter, the resulting mycelia are dried together with the mixed-up remains of the nutrient medium at 20 bio 50 ⁰ C under vacuum or atmospheric pressure, ground and extracted by known methods and the Active substance separated by known oomatographic methods * In the case of the substrates of plant residues of Picea and Pinus, the urea causes an increased growth rate of the mycelia into the substrate. By cli & se operation eliminates the separation of mycelia from nutrient solutions, a setting of the temperatures and the need for sterile operation in the sense of a simple technology for the application of natural waste and

Achieving world-wide constant yields of active ingredient. The formation of psilocybin and the only Ino'ol simplify the separation significantly. The following examples are intended to explain the invention in more detail »

Example: L

From basidiospores of Psilocybe cyanescens a primary culture of 6% solid malt agar is applied, after which an autoclaving mixture of 100 g rice grains and 200 ml water was sterile-inoculated. After 30 days of culture, this inoculum was fibrillated with the spatula after the addition of 100 ml of water, 10 ml of this mycelial suspension were used to inoculate a previously autoclaving mixture of 100 g of Picea dry waste needles, 100 ml of water and 0.3 g of urea, After 30 Days of uneterller fermentation at 15 to 25 ⁰ C was dried at 40 ⁰ C and ground the mass

Dry matter content of Psilocybini 0.28 %

Example 2

With 10 ml of an analogous seed culture of a 2 strain of the same fungal species as in Example 1, 60 g of a natural mixture of fallen leaves, coarsely shredded fallen branches of Tilla and 100 ml of water were added and autoclaved. After 40 days of nonsterile cultivation at 4 to 6 ⁰ C wu <"de dio mixture at 50 ⁰ C in vacuum dried, then zennahlen, exhaustively extracted with methanol and separated on a cellulose column with water-saturated n-butanol, and the individual fractions were tested for Indolderivece with Ehrlich reagent at dor DC, Yield of psilocybin based on the dry product 0y35 %

Claims (2)

claims 1.' A process for obtaining indoles by cultivating agarials, ¹ obtained by solid phase fermentation from Peilooybe oyanescene Wakefield emend, Kriegelsteiner; ¹ characterized by - The cultivation of the fungus mycelia on a solid, natural and aqueous nutrient medium - The cultivating temperature of 2 to 32 ⁰ C. - The cultivation period of 30 to 50 days «Subsequent drying of the fermentation mixture - Extraction and chromatographic isolation of psllocybins. 2Ϊ ^{1 The} method of claim iy characterized in that the drying of the fermentation mixture at 20 to 50 ⁰ C carried out under vacuum or atmospheric pressurev Nutrient medium for the solid phase fermentation of Peilocybe cyanescens, characterized by the composition - fallen branches and foliage of Quercus, Tilia, Salix, Acer, Aesoulud, Populus, Betula, Carpinus, Aldus and Fagus and / or - fallen needles of Picea and Pinus and / or - dead plants of Urtica at the site - Water from 10 to 100 %, based on the solid nutrient medium. 4V nutrient medium according to claim 3, characterized in that the nutrient medium is additionally added with 0 to 01 to 0; 3 % urea. 5. Nursing medium according to claim 3 and 4, characterized dadurchj 'that the nutrient medium is autoclaved before its inoculation and then cooled to a temperature of 2 to 32 ⁰ C.