Claviceps paspali

Lysergsaurederivate kommen besonders im offizinellen Mutterkorn (Sklerotien von Claviceps purpurea) vor. Einfache Lysergsaurederivate werden in beachtlicher Menge auch in saprophytischen Kulturcn von Claviceps paspali gebUdet. 

Arcamone, F., C. Bonino, E. B. Chain, A. Ferretti, P, Pennella, A. To-nolo, and L. Veto Production of Lysergic Acid Derivatives by a Strain of Claviceps paspali Stevens and Hall in Submerged Culture. Nature 187, 238 (1960). 

Kobel, H E. Schreier u. J. Rutschmann 6-Methyl-A > -ergolen-8-car-bonsaure, ein neues Ergolinderivat aus Kulturcn cines Stammes von Claviceps paspali Stevens et Hall. 60. Mitt, iiber Muttcrkornalkaloide. Helv. chim. Acta 60, 1052 (1964). 

The fungus Claviceps paspali Stevens et Hall elaborates a series of metabolites derived from indole and a diterpenoid unit. The structures of paspaline (5) and paspalicine (6) were proposed earlier, together with the partial stereochemistry of paspaline. These conclusions have now been confirmed, and the complete stereochemistry of paspaline and paspalicine has been elucidated, by X-ray... 

From the dried mycel of a Portuguese Claviceps paspali strain, two new indole derivatives, paspalin (1 C28H39N02 mp 264° [a]D —23° in chloroform) and paspalicin (2 C27H31N03 mp 230° [a]D +173° in chloroform) were isolated (3). 

New Alkaloids from Claviceps paspali Stevens and Hall... 

Groger, D., D. Erge and G-H. Floss On the biosynthesis of 2,3-dihydroxy-benzoic acid in submerged cultures of Claviceps paspali Stevens et Hall. Z. Naturforsch. 20 (b), 856 (1965). 

I. S. Kulaev and S. O. Uryson (1965). Free nucleotides and other phosphorus compounds in ergot Claviceps paspali (in Russian). Biokhimiya (Moscow), 30, 282-290. 

Lysergic acid methylcarbinolamide (XXXVI) crystallizes from chloroform in long prisms, mp 135° (dec.), [a]f,° + 29° (c = 1.0 in dimethyl-formamide). The alkaloid, which was isolated together with ergine and erginine from saprophytic cultures of Claviceps paspali (69), easily decomposes in a weak acid solution to form ergine and acetaldehyde. 

Ergonovine and d-lysergic acid N-(l-hydroxyethylamide) occur in some Convolvulaceae and also some strains of Claviceps paspali. Both produce oxytocic action (contractions of the uterus), mydriasis (dilated pupils) and hjrperthermia. 

Arcamone of Farmitalia S.A. Claviceps paspali), and Kobel at Sandoz Laboratories Claviceps purpurea) developed the fermentation of Claviceps fungus for industrial production. 

Isolation of Lysergic Acid from Claviceps paspali Cultures by Werner Schlientz and Benno Sutter Switzerland Januanry 11,1965... 

Example 1-100 Liters of a culture filtrate of the new stock of Claviceps paspali Stevens and Hall [Helvetica ChimicaActa 47,4,1052 (1964)] containing according to the colorimetric analysis 870 mg/liter of alkaloids (relatively has a molecular weight of 268) one adds 800 grams of potassium metabisulfite then 2.5 kg of activated carbon, for example of charcoal or of coal of blood, one agitates the whole during approximately fifteen minutes and one separates by filtration the activated carbon. 

Example 6-100 Liters of a very acid filtrate of culture (pH from approximately 2) of Claviceps paspali Stevens and Hall, whose content, determined by colorimetry, is 1.2 grams per liter (relative with a molecular weight of 268), one adds, while agitating, 500 grams of sodium dithionite and 2.0 kg of activated carbon (Carboraffin). One separates coal by drying, one washes it with 20 liters of water and one subjects it to an elution with 120 liters of methanol added with 10% concentrated ammonia. One concentrates the eluate to 8 liters. One separates by filtration the lysergic acid which crystallized, one washes... 

The previous mothods can be used in the extraction of ergot alkaloids from Claviceps paspali cultures. 

Sclerotia of Claviceps paspali when mature are globular in shape, 2 to 4 mm. in diameter, irregularly roughened on the surface, and yellowish gray in color the interior is homogeneous in structure and contains a considerable quantity of oil. Germinating sclerotia produce... 

Host Plants Resistant to Artificial Innoculation of Claviceps paspali... 

Cultivation of Claviceps paspali Alkaloid Derivatives of Lysergic Acids... 

The organisms employed for the process of the present invention are new strains of Claviceps paspali Stevens and Hall. It had been found that the strains Claviceps paspali Stevens and Hall, which do not produce the lysergic acid derivatives alkaloids by submerged culture, may be virulented artificially, to give new strains of Claviceps paspali, which in turn allow said production. 

The artificial virulentation occurred as follows. Strain F. 97 was isolated from sclerotia grown on plants of Paspalum distichum, collected at Tivoli (Rome) and identified and classified as Claviceps paspali Stevens and Hall. Embryos of Rosen 4 n rye were inoculated, before germination, with the strain F. 97 and then cultivated in vitro. The new virulented subspecies were isolated from sclerotia obtained on said embryos. 

The strains which are used in the process of the present invention and are described as new strains of Claviceps paspali Stevens and Hall in this specification and the claims thereof have been filed at the Istituto Superiore di Sanita, Viale Regina Elena 299, Rome (Italy), and denominated by the marks F-140 F-S13/1 F-237 F-240. The American Type Culture Collection of Washington has assigned to the strains F/s 13/1, F/237, F/240 and F/140 oiClavicepspaspali MllCC numbers 13892, 13893,13894 and 13895, respectively. 

The process of the invention is therefore one for the production of alkaloid derivatives of lysergic acid which comprises fermenting under aerobic conditions an aqueous nutrient medium containing a source of carbon, nitrogen and mineral salt with a new strain of Claviceps paspali Stevens and Hall as herein before defined. 

As regards the production of alkaloid derivatives of lysergic acid, the present invention is not limited to the use of the described strains, but comprises also the mutants thereof, which may be obtained, e.g., by means of either a selection or a mutation by the action of U.V. rays or Roentgen rays or any other mutagenous substance or, particularly, by artificial infection of either embryos or grasses cultivated in vitro or plants of grasses cultivated both in vivo or in vitro and the said mutants are to be included in the definition of a new strain of Claviceps paspali Stevens and Hall. 

The process is carried out in 500 mL. flasks containing 100 mL. of a suitable nutrient medium. The flasks are shaken by a rotary shaker (200 revolutions/minute eccentric throw 10 cm.). The optimal incubation temperature is at 27° C. The relative moisture is 85-90%. The cultivation is carried out in the dark. A flask is inoculated with the mycelium which is obtained from a 10 days culture in potato-glucose-agar of one of the above-described new strains of Claviceps paspali Stevens and Hall. The nutrient medium is the following ... 

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