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OfflineNalim
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Ergot cultivation
    #6607718 - 02/25/07 05:30 AM (7 years, 7 months ago)

I have some info I thought might be of interest for some of the people here.

This is taken from the book Pharmacognosy; Phytochemistry, Medicinal plants

Industrial Fermentation

The different Claviceps can be cultivated at pH's near 5.5, which are best adjusted with ammonia salts of the acids in the Krebs' cycle(succinate, citrate).
Frequently the alkaloid production is dependent on the phosphate concentrations of the medium, and for many strains, on the precise concentrations of minerals(iron, zinc, copper, boron) which directly influence the productivity.
The oxygenation of the culture medium must be intense.
Cell differentiation and alkaloid production are controlled by the addition of nutrients: a rich medium causes the formation of abundant hyphal mycelium and no alkaloid production. The latter is triggered when the medium becomes poor in certain nutrients. For many strains, it is the phosphate concentrations that conditions the passage from mycelium proliferation to the biochemical differentiation phase.
Tryptophan, added at the beginning of the fermentation, acts as an enzymatic inducer, and therefore increases the quantity of alkaloids formed.


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OfflineNalim
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Re: Ergot cultivation [Re: Nalim]
    #6607739 - 02/25/07 05:51 AM (7 years, 7 months ago)

Alkaloids produced

Two approaches are possible. The first one consists of producing simple ergolenes, namely paspalic acid or the hydroxyethylamide of lysergic acid. These two compounds are biosynthesized by Claviceps paspali Stephens and Hall which yields largely exceeding 2g/L. Other calviceps could potentially produce clavines with much higher yields.

Once isolated from the culture medium, the two ergolene derivatives are transformed-the first by isomerization, the second by hydrolysis-into lysergic acid, the starting material for the synthesis of non-peptidic alkaloids used in therapeutics(ergometrine and others). Some clavines can also be the starting material for the synthesis of more complex products.

The second approach, which is more recent, leads to the direct production of ergopeptines, particularly ergocryptine from strains of C. purpurea and possibly from other species. The fermentation is lengthy, and the ergopeptines are obtained in a yield in the order of g/L. The culture can be directed by adding to the medium the amino acids that are the precursors of the desired tricyclic peptide.

Since the specificity is low it is possible to introduce in the medium abiogenetic amino acids, in order to obtain alkaloidal compounds that do not occur naturally and are novel for pharmacological experimentation.
In any event the the products must be extracted, keeping in mind that a fraction remains inside the cells(ion-exchange resins).




If anyone has any links related to the subject of cultivating ergot this is the place to post them.
Would be great if anyone has a recipe for the nutrient solution.


--------------------

Rodney Brooks on Robots


Edited by Nast (02/25/07 06:05 AM)


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OfflineSofaKingGood
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Re: Ergot cultivation [Re: Nalim]
    #6607859 - 02/25/07 08:48 AM (7 years, 7 months ago)

Yes please if anyone knows the recipe for the nutrient please share. On Monday I'm going to go talk to an old chemistry teacher and find out if he knows anything about ergot cultivation or how to obtain some of the acids in the first technique.


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OfflineNalim
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Re: Ergot cultivation [Re: SofaKingGood]
    #6607916 - 02/25/07 09:39 AM (7 years, 7 months ago)

you should be able to prepare the ammonia salt of citrate trough mixing Ammonium hydroxide(most online chem-suppliers) with citric acid(any food store on the spice shelves) if I'm not mistaken... Just make sure you use equimolar amounts.


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Offlineshroomsynergy
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Re: Ergot cultivation [Re: Nalim]
    #6608027 - 02/25/07 10:49 AM (7 years, 7 months ago)

Tricarboxilic Acid (i think that is how you spell it...) is an acid in a kelp base that will drastically increase the Krebs cycle given the right dosage at the right times in plant growth. i would think that would cause a hightened alkaloid production. I use it on my buds and they get nice and sticky-icky. i use it during cloning, then just after the transfers from one cycle to the next. You can get it at select garden centers and hydroponic shops.

i know thats not your question, but it may help.


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OfflineNalim
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Re: Ergot cultivation [Re: shroomsynergy]
    #6608144 - 02/25/07 11:49 AM (7 years, 7 months ago)

I think it's pretty hard to predict how the fungi will react to a given substance unless you actually try it.
The reason you use ammonium salts of the Krebs cycle acids is probably because you don't want to add any minerals that might affect the alkaloid production or the growth of the fungi.
Nevertheless if one where to cultivate ergot successfully, substituting the succinate or citrate salts with the ammonium salt of tricarboxilic acid might very well prove to beneficial to either growth or(much more likely) alkaloid production.


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OfflineXtals
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Re: Ergot cultivation [Re: Nalim]
    #6609098 - 02/25/07 05:38 PM (7 years, 7 months ago)

There are different ways of getting ergot to produce different alkaloids, though of interest to the LSD chemist are ergotamine, ergotaminine and ergometrine.

Try using the European Patent Office's search function on their website. I have plenty of patents and academic journals (some of which you can obtain for free through Pubmed Central) that I would be happy to share, when I get access to my files (they aren't on this computer).

The European Patent Office's search function can be accessed through:

http://www.espacenet.com/

(chose your gateway country and then search)

PubMed Central (which gives full access to free articles) can be found:

http://www.pubmedcentral.nih.gov/

Just for a start, take a look at US2809920 (that's US Patent #2809920) which you can find by chosing "number search" and typing in "US2809920." The title is: Process for the preparation of ergotamine, ergotaminine and ergometrine by saprophytic culture of ergot (claviceps purpurea [fr] tul.) in vitro and isolation of the alkaloids thus produced

After clicking on the "original document tab" at the top you'll see an option for "save full document" which allows you to download it as a PDF.

The academic journal articles I have on this subject agree with the patent literature (and generally, it would be stupid for someone to try patenting a process that didn't work - what, exactly, would be the point in that? Nobody would use the process so you'd have nothing to gain). For this kind of thing, patents are better at giving actual practical examples, while the academic literature is good for giving and explanation of what the investigators were looking at (and sometimes the "materials and methods" sections aren't too helpful).

On Pubmed Central, just for starters, search for Biosynthesis of ergotamine by Claviceps purpurea and you'll find a 1.3 Mb PDF by Basset, Chain and Corbett from 1973. Also take a look at Ergotamine Production in Submerged Culture and Physiology of Claviceps purpurea. The latter article gives references for some media referred to like media T2, media T25 and media TG.

I find Google Scholar is pretty good but it doesn't necessarily bring up articles that you actually can access.


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OfflineNalim
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Re: Ergot cultivation [Re: Xtals]
    #6609285 - 02/25/07 06:28 PM (7 years, 7 months ago)

:laugh::thumbup:
Thanks! Will be reading those articles tomorrow.


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OfflineCerius
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Re: Ergot cultivation *DELETED* [Re: Nalim]
    #6610393 - 02/25/07 10:54 PM (7 years, 7 months ago)

Post deleted by RogerRabbit<p>Reason for deletion: Cleaning up thread so I can re-open it.


Edited by RogerRabbit (02/28/07 07:07 AM)


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OfflineNalim
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Re: Ergot cultivation [Re: Cerius]
    #6612101 - 02/26/07 12:53 PM (7 years, 7 months ago)

I know there are alternatives, they have been(/is) discussed in ODD..
They do not belong in advanced mushroom cultivation though.
I know that ergot is toxic and that it might not be the "best" way to do it, but it is a way to do it.

Yoschie99 posted this in the LSD thread, seems Ythan beat me to it.:D

The recipe seems easy enough, and getting claviceps is not that hard, it is a pretty common fungi in most of the world, and who are more well equipped to id and isolate it than the members of this forum?


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Offlineillahee
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Re: Ergot cultivation [Re: Nalim]
    #6613242 - 02/26/07 06:18 PM (7 years, 7 months ago)

I know that ergot is very common, however I think that finding a strain that will produce alkaloids in a petri dish is difficult. This is what I've heard on the subject before. Perhaps it isn't a question of the right strain but of the right nutrients.


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Invisiblefastfred
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Re: Ergot cultivation [Re: Nalim]
    #6613258 - 02/26/07 06:23 PM (7 years, 7 months ago)

Ergot is the way to go. You're not going to get ergotamine any other way without having the DEA kicking your door in in short order.

Isolating a high yielding strain is going to be the hard part. According to the literature there is over 2000X variability in the alkaloids produced. If you have some connections the best way would be to order one of the high yielding strains from ATCC (American Type Culture Collection).

Otherwise it would be pretty tough to analyze each culture and find a high yielding isolate.


-FF


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Invisiblefastfred
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Re: Ergot cultivation [Re: fastfred]
    #6613386 - 02/26/07 06:59 PM (7 years, 7 months ago)

> Perhaps it isn't a question of the right strain but of the right nutrients.

Nope, it's all about the genetics. I'm sure there is some variability based on the nutrients, but it is dwarfed by the genetic aspect.

RE: My last post, I noticed that ATCC doesn't list any Claviceps cultures. Your best bet would be to look up some papers and find where they deposited their voucher specimens.

Here are some references for you...

Interesting Chemical Abstracts References:
CA: 71:P69351y : describes mutants of strain NRRL 3027
producing 2270mg/l of ergoline compounds of which 85% were
amides of lysergic and isolysergic acids. (Swiss patent application)

CA: 77:P156333n : describes a method used to get 9.75g pure
lysergamides from a 10 litre fermentation originally containing
an estimated 15g.


A note on the variability of alkaloid content of domestic ergot.
Atal, C. K.; Schwarting, A. E. (1957)
J Am Pharmaceut Assoc Sci Ed 46:451

Studies on cultural requirements of Claviceps purpurea and inactivation of ergotamine.
Michener, H. D.; Snell, N. (1950)
Am J Bot 37(Jan):52-59

Physiological diferentiation of alkaloid producing strains of Claviceps purpurea
Authors: Matošiæ, Sreæko; Mehak, Milena; Šuškoviæ, Jagoda
Journal: Acta Bot. Croat.
ISSN: 0365-0588
53, pgs.21-30 (1994)
Summary: Four different strains of Claviceps purpurea IC/39/20-B, G, R and W have been selected and isolated using common selection method. The relationship between intensity of pigmentation of the culture and the accumulation of ergot alkaloids have been noticed. Maximal alkaloid yields have been obtained with more pigmented strains R and W in the sucrose-asparagine medium (1.30 and 1.50 g/L respectively) and with less pigmented strains B and G in the sucrose-peptone medium (0.30 and 0.80 g/L).



-FF


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OfflineXtals
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Re: Ergot cultivation [Re: Cerius]
    #6613838 - 02/26/07 09:15 PM (7 years, 7 months ago)

Quote:

Cerius said:
Ergot cultivation is a dangerous and probably a fatal hobby.




Is it really? Can you back this up? Do you have any articles from newspapers, magazines, scientific journals about accidents involving ergot?

Ergot is a fungus, like P. cubensis. Ergot cannot walk. Ergot cannot fly. You handle poisonous substances everyday. As long as you are careful not to eat the ergot, you will not die from it. All one needs to do is be careful and wear gloves and avoid eating the fungus.

Thank you for further contributing to the mythology. There are industrial producers of ergot that grow submerged cultures by the thousands of liter. Other industries intentionally infect fields of rye with it and go out and collect it. Unless you accidentally swallow an ergot or take a bath in the extract you have nothing to worry about.

Why is it that people have to make up this kind of nonsense? Ergot cultivation is not "probably" a fatal hobby. I am so sick of the mythology that is grounded in absolutely nothing.

Quote:

You know it's poisonous, right?




So is Aspririn. Why don't you worry about drying from that? Rubbing alcohol is poisonous too. So is bleach. All you have to do is avoid swallowing these things. Ergot is an immobile fungus. Wear some latex gloves and don't eat it. It can't fly. It can't walk.

Quote:

If you're looking for precursors to LSD-25, there are plenty of chemists who spend there lives doing this for you.




No. Chemists do not sit around making precursors for illicit LSD manufacture.

Quote:

There's are "other" alkaloids besides ergotamine that are easily and readily available;




Name them and their sources.

I'd like to hear this.

Quote:

A grignard reagent and you're pretty damn close to the wet dream, baby.




At what step in an LSD synthesis would you use a grignard? If you answer this, please answer it in the ODD forum in the "why is acid so hard to make" part. This thread isn't about LSD synthesis, really, as we have a huge thread going about this. Even if we don't have intent on or even interest in manufacturing LSD (as I don't particularly), the question of why ergot is, how it grows, how it synthesizes alkaloids is still interesting.


Edited by Xtals (02/26/07 10:11 PM)


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OfflineCerius
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Re: Ergot cultivation *DELETED* [Re: Xtals]
    #6614224 - 02/26/07 10:38 PM (7 years, 7 months ago)

Post deleted by RogerRabbit

Reason for deletion: Cleaning up thread so I can re-open it.



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Offlinejccc
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Re: Ergot cultivation [Re: Xtals]
    #6614252 - 02/26/07 10:44 PM (7 years, 7 months ago)

http://en.wikipedia.org/wiki/Ergot Read the article on affects on humans...now from what ive heard inhaleing anything related to ergot doesnt matter its when you ingest the wrong stuff that your life goes to hell. http://www.erowid.org/plants/ergot/ergot_cultivation1.shtml Sorry if this was posted before ddnt have timeto read the articles but his is very simple and easily obtainable materials!!


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OfflineXtals
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Re: Ergot cultivation *DELETED* [Re: Cerius]
    #6614457 - 02/26/07 11:17 PM (7 years, 7 months ago)

Post deleted by RogerRabbit

Reason for deletion: Cleaning up thread so I can re-open it.



Edited by Xtals (02/26/07 11:35 PM)


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OfflineXtals
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Re: Ergot cultivation [Re: Xtals]
    #6614664 - 02/26/07 11:57 PM (7 years, 7 months ago)
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Sorry about that everyone, but I guess I just had to deal with that.

Again, ergot cultivation CAN be dangerous but it doesn't have to be. One needs to be aware that ergot alkaloids are very poisonous and accidentally swallowing an ergot could be a death sentence. However, people are generally pretty good at avoiding swallowing things that they don't intend to, so if you do find ergot in the field and intend to look at in under a microscope or something, it would be best to wear latex gloves when handling it, or use a pair of tweezers. If ergot did have special properties, like its spores were also poisonous and could be inhaled and might kill you, then you'd probably be able to find that information very easily. That's the kind of thing that gets reported and I've never run across anything like that.

Also, if you have other suggestions for handling it safely, please make those suggestions here.

Personally, I'd be interested in seeing so microscopic pictures of it from recent field samples.

I'm a sucker for Stoll and Hoffman's work, so here's a patent by them, about preparing ergotamine, etc. from C. purpurea.

I think biosynthetic investigations are also neat and would like to discuss those.


Edited by Xtals (02/26/07 11:59 PM)


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Re: Ergot cultivation [Re: Xtals]
    #6615982 - 02/27/07 10:39 AM (7 years, 7 months ago)

....


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Peace

I get real lonely


Edited by Tamadragon (02/28/07 04:25 PM)


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Offlineillahee
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Re: Ergot cultivation [Re: Tamadragon]
    #6616100 - 02/27/07 11:56 AM (7 years, 7 months ago)

More props to Xtals! thank you for your very well voiced opinion in this matter, it's nice to see a smackdown here and there.

Tamadragon, you have no idea of what some people are capable of that surf this forum. Literally, you honestly have no fucking idea at all....and I really mean, AT ALL!

Personally, if I found out that a member had a strain of ergot that could produce alkaloids in a dish i'd hunt them down and beg for a sample.

Xtals, have you read up on the L-PAC biosynth of ephedrine alkaloids? Feed yeast some benzaldehyde and they spit out alkaloids.


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