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Hexabromobenzene
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Production of butyric acid acid and butanol by fermentation
Production of butyric acid and butanol by fermentation. This article was written with google translate. If you have any questions write in the topic
Summary. Butanol and butyric acid can be obtained by fermentation starch сontaining raw by clostridium sp.
Clostridium strains can be obtained from potato peels submitted thermal shock in hot water for 10 minutes and further cultivation on starch raw
materials with source of calcium magnesium and nitrogen
Concentration carbohydrates in fermented liquid must be not be more 50-60 gr liter solution if butanol may be obtained. In case butyric acid
fermentation in liquid must be added about 700gr calcium carbonate per 1 kg carbohydrates. Cabohydrates concentration may be up to 100 gr liter.
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This article explains how at home to establish the production of butyric acid and butyl achohol by fermentation. It is divided into 4 parts.
1. Theory
2. Purification bacterial strain
3. Preparation of raw materials and its fermentation
4. Processing of fermented liquid to the required products
1. Theory
In nature, there is butyric acid fermentation caused by various types of bacteria (as a rule of Clostridia). Butyric Acid is obtained from the
condensation of two acetyl-cola molecules and accumulates in solution. Butyric acid has a very unpleasant smell resembling rotten socks, rotten
potatoes, toilet, cheese blaver and the like (depends on the tastor).
Some types of clostridium when accumulating butyric acid change their metabolism and are obtained by neutral products such as acetone, butanol, IPA
(depends on the strain). They are found in the ground, but the best source of potato peel Some straps of clostridium can produce only butyric acid
(they do not have butyraldehyde dehydrogenase and butanol dehydrogenase). Their common name is Clostridium Butyricum. Stamps capable of producing
butanol have a common name for Clostridium Acetobutylicum
When obtaining butyric acid, it is necessary to neutralize it with for example calcum carbonate. Acid medium toxic for clostridium. In this case, the
equilibrium will be shifted towards acidic extensions (obtained in the form of salts). In the production of butanol, when it accumulates in above
10-15 grams, a liter of clostridium stops life and form disputes. Clostridy belongs anaerobic. Produce spores produce that are extremely chemically
resistant. This property is based on this method isolation. Unlike yeast, they are capable of growing without access of oxygen. They are much easier
to breed and do not buy new bacteria for cooking new fermented liquid
2. Purification bacterial strain
As a source of spores of bacteria, it is best to use potatoes peel. Not every potato will go. You may have to try to one. As a cultivation
environment, you can use oatmeal. This technique Oatmeal boiled with a calculation of 60-70 grams of carbohydrates per liter of fermentation liquid in
porridge Add gram 2 per liter of Technical calcium carbonate (mixture of calcium carbonates and magnesium for better growth) Next, the peel of
potatoes peel poured with water temperature degrees 90 and keeps it for 5-10 minutes.
For the first time, it is better not to take a substrate much (in case of failure it will be hurt). After treating potatoes with water, we fill this
mixture into our cold substrate(Oatmeal) already in the fermentation container (for example, a 2 liter PET bottle). To protect our clostrid from the
air there are 2 options. Either pour with vaseline oil or sunflower oil (pasteurized) or to outweigh the gas from the bottle with gas butane for
lighters and immediately close it and attach the hydropout. I recommend the second option as oil trails will then be everywhere when using ready-made
biomaterial.
And so after all the preparations in a few days, fermentation should begin and our raw materials can float up it is normal. Now it remains only to
wait (from week to month). As fermentation, the smell changes with an unpleasant butyric acid on the smell of glue and then such a fruity (banana,
pineapple and so on). If the smell began to change to glue(butanol) and fruity. This is a success! You are isolated the desired strain
In theory, the starch raw materials can already be infected with not fully fermented liquid, but I recommend waiting for the end of fermentation and
take the liquid from the bottom of the fermentation container with the residues of the croup. There, the highest concentration of the clostridia
spore and 10-20 ml is enough for clock through 10-20 to cause began fermentation of 30 liters of fermention liquid. Further fence of culture can be
taken from the bottom of the ready-made fermented liquid or their filtrates that are further recycling.
3. Preparation of raw materials and its fermentation
Preparation of fermentation liquid to fermentation similar to the previous stage, but a little different This is done to reduce the cost and simplify
the preparation of a large number of fermented product. To begin with, we need to decide what we need. Butyric acid or butanol with acetone.
Conditions for their preparation are different. For butyric acid, we must add about 500-700 grams of Technical calcium carbonate to fermentation
liquid (technical mixture of carbonate hydroxides calcium magnium) per kilogram of carbohydrates. The permissible concentration of carbohydrates may
be probably increased up to 100 grams per liter for butyric acid fermentation. Fermentation of butyric acid goes without inhibiting full and to the
end. For fermentation butyric acid, it is also possible to use strains that did not give adhesive and ether smell (keeping some strains are produced
in addition to butyric acid very smelly products. Probably indole and skatole)
If we want to produce a butanol and acetone (some strains produce only Batanol some IPA instead of acetone), calcium carbonate need about gram 2 per
liter. Bacteria needs magnesium iron and calcium that are present in the technical calcium carbonate.
Carbohydrates need not greater than 60 grams per liter due to the fact that the produced butanol very much inhibits fermentation. Even with such a
loading at the end, fermentation is very slow.
Sugar can be used as carbohydrate raw materials.
You can use starch raw materials as black and white bread, various cereals, starch, potatoes. Before use, starch raw materials should be pouring
boiling water and hold at least 40 minutes for swelling and the destruction of microorganisms. With flour of pasta and boiled batches, fermentation
goes very quickly. Slowly with cereals of rice buckwheat. The grains slowly destroyed at the bottom of the fermentation container converting with such
a mass and rarely float up. Pea cereals and solid grain wanders very badly Potatoes cut by pieces and everything apparently does not ferment
everything.
Probably from him must be done. The larger the cereals and starch raw materials the greater you need to mess around with them.
Here is a ready technique
2 kg of old slightly molded banets were filled with 10 liters of boiling water and left 2 hours before turning into porridge. Further, this porridge
was loaded along with 700 grams of calcium carbonate and a Diluted hot water from under the tap (60 degrees) to 20 liters. Residues from the bottom of
a other fermentation container where butanol was obtained (about 100 ml of a culture sample) entered in the new fermentation container. Further, all
this is stirred, butane gas was launched in the fermentation container. The container was closed with a lid and the gauge slog was connected to the
hydraulic assembly. Fermentatiom began after 12 hours. Porridge from banets surfaced to the top. Usually this porridge dissolves under the end of
fermentation. In theory, butyric acid should be obtained as the calcium carbonate was added. Similarly, you can use any food waste After fermentaion
liqid was filtred. The sediment from the bottom of the container can be used for new fermentation. This is a biomaterial. Our bacteria that can then
use it. Can be stored up to year If you use such raw materials like sugar, then you will probably need to add a nitrogen source as urea And so when
the gases stop out, the starch raw materials (if used) was completely dissolved and an ether smell appeared (in the case of butanol. Although it also
happens at the end of butyiric acid fermentation), then fermentation is considered to be completed. Next, the stage begins to highlight products from
fermented liquid
4. Processing of fermented liquid to the required products So. After fermentation of fermented liquid, we have a solution of calcium butirate or
acetone mixture and butanol in water with any suspension. First, fermented liquid must be filtered and drained from the sludge at the bottom of the
fermentation container. You can filter through sheets or bag of stitched polypropylene. In some cases, fermented liquid on consistency is similar to
snot probably because of the granoluse or remnants of gluten with flour (definitely not clear). With such a material it is very inconvenient to work.
After filtering, the sludge leave it contains our spores bacteria Fermented liquid that we filtered should be Frozen and liquid flow from ice. In
such a concentration, there is no particular sense to mess with further processing of fermented liquid. It will be very energy consuming Fermented
liquid contains calcium butyrate mus evaporated after freezing Butanol and contaning fermented liquid must be distilled
In the case of butyric acid fermentation, it is enough to evaporate it (carefully! Strong smell), to percipate the salt and then add inorganic acid
and separated by distillation butyric acid azeotrope. Solubility of calcium butirate is about 300 grams per liter. Butyric acid always contains after
fermentation impurities of acetic acid, which must be separated In the case of Acetone, Butanol Fermented liquid needs fractional distillation after
freezing. At first, at 50-80, a mixture of acetone and water is distilled, then ethanol or IPA (depends on the strain) at 80-92 and then the butanol
azeotrope containing it is about 20% . Some strains produce only Butanol. |
[Edited on 26-6-2021 by Hexabromobenzene]
[Edited on 26-6-2021 by Hexabromobenzene]
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draculic acid69
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Do U plan on synthesizing this abomination yourself or are U just throwing it out there for us to see?
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karlos³
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I didn't know Clostridium cultures were that easy to obtain; by random coincidence I just read about a species of them yesterday 
@mods, could you please move that to the biochemistry section?
It fits much better there.
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Hexabromobenzene
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I did it. It's work.
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Hexabromobenzene
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I prepared 50 liters fermentation liquids (20 liters butyric acid and 20l butanol)
Here is instructions(Russian):
https://youtu.be/UbRxmkBSIx4
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Triflic Acid
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Why are you biosynthesizing butyric acid like this? There is a much better method that requires no chemicals and also produces hydrochloric acid. Just
go eat some moldy cheese and you'll get a lifetime supply of hydrochloric and butyric acid on tap 
There wasn't a fire, we just had an uncontrolled rapid oxidation event at the power plant.
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draculic acid69
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Quote: Originally posted by Triflic Acid  | | Why are you biosynthesizing butyric acid like this? There is a much better method that requires no chemicals and also produces hydrochloric acid. Just
go eat some moldy cheese and you'll get a lifetime supply of hydrochloric and butyric acid on tap |
Hahh!!! Nice one
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Tsjerk
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Clostridium forms spores which survive pasteurization, and practically all potatoes are contaminated. So if you pasteurise for about five minutes you
will have highly enriched your sample with Clostridium.
Nice article: https://pubmed.ncbi.nlm.nih.gov/3290178/
Even the negative controls were contaminated 
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Hexabromobenzene
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However it's works. Butyric acid and butanol strain was prepared from different potatotes. About 50l fermentation liquid was prepared. No one botulin
toxin poisoning was observed. Sometimes fermentation liquid taken by naked hands
Best results give potatotes, which have a characteristic smell
Clostridium botulinum needs proteins medium
[Edited on 30-6-2021 by Hexabromobenzene]
[Edited on 30-6-2021 by Hexabromobenzene]
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Tsjerk
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I didn't read carefully, but I was under the impression the fermentation is done by Clostridium, so you need them to make the process work.
Also, the toxin is heat sensitive, so as long as you are careful with the raw fermentation, you should be fine.
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Hexabromobenzene
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The following materials were used to fermentation: starch, boiled potatoes, Boiled wheat, rise, peas,oatmeal, moldy bread, wheat flour, millet,
buckwheat, other food waste,
sugar(Attention! Other bacteria in crude culture and raw matherial can fermentate it! Lactic acid as example toxic for clostridia)
starch containing raw matherial need to treat hot water (100C) about hour
[Edited on 1-7-2021 by Hexabromobenzene]
[Edited on 1-7-2021 by Hexabromobenzene]
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Hexabromobenzene
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3.5-4 kg old dried bakery products wastes was spilled hot water(100с) in 20 liter bucket and left for 40 minutes. This mixture was placed in a 40
liter fermenter. 2 kg of calcium carbonate and hot water (60C) was added to the mixture to a volume of about 40 liters. About 300 ml of mass is added
to this mixture from the bottom of the past fermentation (bacterial culture). Bacteria was isolated from potato peel. Gas propane was added to the
fermenter for air displacement. Active fermentation began after 12 hours and continued until 2 weeks. During fermentation there was heat (about 30
degrees) and fermentation ended 2-3 times faster than at 20 degrees
After a month, the fermentation liquid was clear and the precipitate from the calcium carbonate and the spore of the bacteria fell to the bottom.
Bakery products was completely dissolved
Fermentation goes in 2 stages. During the first stage, starch is fermented and butyric and acetic acids are formed with gases. During the second
stage, proteins are fermented. With these, smelling compounds such as skatole, mercaptans, disulfides are formed. It is also formed possible
isovaleric acid and caproic acid.
The liquid was decanted from the sediment and frozen in the freezer in small portions. After freezing, the liquid separated from ice. from 35 liters
fermentation liquid 7 liter concentrate was obtained. Concentrated liquid was extracted with 500 ml toluene(xylene also may used) to remove
unpleasantly smelling compounds(skatole main). Extracted liquid was evaporated(WARNING!!! VERY STRONG VOMITING SMELL!!!). After evaporation, 1300
grams of wet(About 30% water) crude salts of carboxylic acids were obtained (acetic and butyric). The total yield of acids is not more than 600-700
grams
Hydrochloric acid was added to the mixture but only 100-200 dark brown ml of liquid floated upper layer. Calcium chloride is a bad substance for
salting off acids. The resulting mixture should be distilled but it has a very unpleasant smell.
[Edited on 13-11-2021 by Hexabromobenzene]
[Edited on 13-11-2021 by Hexabromobenzene]
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macckone
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The smell of vomit is the smell of butyric acid.
Adding some steel wool should reduce the amount of sulfur compounds formed.
Copper will also remove sulfides.
Apparently c. acetobutylicum are not sensitive to copper and it supposedly improves yields of butyl alcohol due to increased hydrogen production.
Will have to find the correct reference.
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j_sum1
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So. It can be done. But I think I will buy any butyric acid that I need.
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Hexabromobenzene
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Fermentation gives not only butyric acid. Small quantity skatole may be also obtained. And butanol with acetone can obtained without calcium
carbonate.
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unionised
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Have a quick look at this, then see how man of hem you want in your home...
https://en.wikipedia.org/wiki/Clostridium
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Hexabromobenzene
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Starch fermenting strains non pathogenic. Fresh culture may be contaminated, but pathogenic strains lose pathogenicity and displace in natural
selection by other strains. Are you not afraid of rotten potatoes?
[Edited on 13-11-2021 by Hexabromobenzene]
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macckone
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If you have raw potatoes in your home you have the ABE bacteria that is cultured.
They are ubiquitous.
There are pathogenic variants, but they aren't likely to be on your potatoes.
Or at least I would hope they aren't.
The clostridium are very hardy and the shock boiling step selects the hardiest.
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Tsjerk
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| Quote: Originally posted by Hexabromobenzene | Starch fermenting strains non pathogenic. Fresh culture may be contaminated, but pathogenic strains lose pathogenicity and displace in natural
selection by other strains. Are you not afraid of rotten potatoes?
[Edited on 13-11-2021 by Hexabromobenzene] |
Bullshit, all Clostridium botulinum strains are "pathogenic". Although that depends on your definition of pathogenic. Pathogens are organisms or
viruses that can infect, which this "strain" can't, but it definitely produces botuline which is very toxic. Am I afraid of rotten potatoes? Very
definitely, when they are not heated to 100 degrees for at least 20/30 minutes. I put strain between quotes because we are not talking about a strain
but a species.
Is all this a problem? No, I don't think so, because you won't drink the fermented liquid, and when processing you heat everything above the
temperature at which botuline is denatured. But I still think calling the "strains" you isolate "non-pathogenic" naive and irresponsible.
There is a reason this fermentation smells as bad as it does, and that is because our noses evolved to recognize the extreme toxicity it contains.
Also working with fermentations containg enough botuline to supply the world's cosmetic medical demand for at least a year without knowing it does is
dangerous in my eyes, to say the least.
Whatever you isolate from potatoes will be, or at least you should assume without running 16S PCR, Clostridium botulinum. And that will produce
botuline. Almost all Clostridium species are bastards. Look at botolinium, difficile, tetani for example.
[Edited on 14-11-2021 by Tsjerk]
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Hexabromobenzene
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Clostridium botulinum does not fermented starch. Also, I'm not sure if it can be fermented gluten. More likely no than yes. They need specially
prepared proteins
Under the pathogen, I understand the ability to infect a person
It is possible dangerous to hit the fluid in deep wounds. If superficial are not dangerous if you wash and process the antiseptic (verified)
My cat licked a little (less ml of liquid) but still alive without side effects
[Edited on 14-11-2021 by Hexabromobenzene]
[Edited on 14-11-2021 by Hexabromobenzene]
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Tsjerk
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| Quote: Originally posted by Hexabromobenzene | Clostridium botulinum does not fermented starch. Also, I'm not sure if it can be fermented gluten. More likely no than yes. They need specially
prepared proteins
Under the pathogen, I understand the ability to infect a person
It is possible dangerous to hit the fluid in deep wounds. If superficial are not dangerous if you wash and process the antiseptic (verified)
My cat licked a little (less ml of liquid) but still alive without side effects
[Edited on 14-11-2021 by Hexabromobenzene]
[Edited on 14-11-2021 by Hexabromobenzene] |
Your cat licking is not a good indication.
Did you do a clean streak? When you didn't, you are growing Clostridium botulinum
[Edited on 14-11-2021 by Tsjerk]
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S.C. Wack
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| Quote: Originally posted by Tsjerk | | There is a reason this fermentation smells as bad as it does, and that is because our noses evolved to recognize the extreme toxicity it contains.
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...dubious...and, animals have no such qualms, so they must be immune?
BTW Clostridium butyricum is sold OTC as a probiotic.
So sour milk or moldy cheese or fermented starch is where botulism comes from? Exactly which cases can be traced to butyric fermentation.
There are of course details in the old lit, which fail to mention any danger, because that's how n-butyric acid and things made from it came to be
commercially available...B. subtilis from washed hay was also recommended, no doubt some disease caused by it will be brought up...
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Tsjerk
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Okay, botulinum doesn't fermet starch.
https://microbenotes.com/biochemical-test-of-clostridium-bot...
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karlos³
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It does smell bad because of the butyric acid, I thought?
I worked with n-butanal just a few days ago... I think it had that pungent smell because of tiny amounts of butyric acid dissolved in it, no idea.
But it smelled like someone puked over a flower patch.... but one of the not so nice kind, likes the little Tagetes we plant under tomatoes as natural
nematode repellent.
Yeah, that was the smell of n-butanal.
Just a fraction of butyric acid though.
verrückt und wissenschaftlich
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walruslover69
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Back in my younger days I attempted to make some moonshine out of corn. I left a one gallon batch ferment way too long (maybe 3 months), and attempted
to distill and salvage it. After I distilled over most of the ethanol, The water that started coming over was absolutely putrid, strongly smelling of
vomit. I'd know how much, but a surprising amount of butyric acid seemed to be produced. I considered isolating the butyric acid, but it was far too
unpleasant to work with under my lab conditions at the time.
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