Sciencemadness Discussion Board - Citric acid

Citric acid

Waffles SS - 22-1-2011 at 07:35

Quote:

In 1917, the American food chemist James Currie discovered that certain strains of the mold Aspergillus niger could be efficient citric acid producers, and Pfizer began industrial-level production using this technique two years later, followed by Citrique Belge in 1929.

In this production technique, which is still the major industrial route to citric acid used today, cultures of Aspergillus niger are fed on a sucrose or glucose-containing medium to produce citric acid. The source of sugar is corn steep liquor, molasses, hydrolyzed corn starch or other inexpensive sugary solutions.[10] After the mould is filtered out of the resulting solution, citric acid is isolated by precipitating it with lime (calcium hydroxide) to yield calcium citrate salt, from which citric acid is regenerated by treatment with sulfuric acid.

Anyone has experience on making citric acid?
Where we can buy Aspergillus niger?

[Edited on 22-1-2011 by Waffles SS]

bbartlog - 22-1-2011 at 09:53

There are presumably commercial cultures for sale that are optimized for citric acid production (i.e. specific strains of aspergillus niger), but I don't know who sells them. Easier if you just want to experiment is to find an onion with black mold on it; this is highly likely to be aspergillus niger.
Searching google books for 'aspergillus niger culture' will turn up a lot of early 20th century works that give guidelines on culture medium and other conditions. Unless you're good at maintaining sterile conditions you will want to lower the pH of your medium to 2-2.5, since aspergillus niger is one of the few organisms that can grow in such acidic conditions. Scrape black mold off onion into the medium, see if anything grows.

Waffles SS - 24-4-2011 at 00:16

I separate two black mold from two onion and provide sterile condition for aspergillus niger also i control ph between 2-2.5 .now i want to make sugar solution and put aspergillus on it.
but i have a little information about it .what temperature ?what condition?really i dont know
i searched in google and i didnt find any useful information
my friend decide to buil small company for making citric acid by this method but really even we dont know where we can buy aspergillus niger

Steve_hi - 24-4-2011 at 04:38

Hi waffles is this any help to you
http://www.carolina.com/product/fungi%2C+basic+mold+showplat...

Carolina biological supplies.
I think I'll try to cultures some A.niger

Saerynide - 24-4-2011 at 08:33

I've never cultured A. niger, but I have cultured other fungi and a temp of 70-80F seemed to be ideal. The mycellium I had grew very fast (and so did contamination when I wasn't careful).

Interesting that you'd like to grow A. niger, because the I would throw the whole thing out like it was a hand grenade without a pin if I saw a speck of black mold. I heard a lot of very scary stories from my mycology professor about Aspergillus species (ie: Aspergillosis, aflatoxin...)

bbartlog - 24-4-2011 at 13:49

Check out 'The citric acid fermentation of Aspergillus Niger', by James Currie, in 'The Journal of Biological Chemistry', Volume 31 (1917). It's available on Google Books. It should provide you with a wealth of information, including what they (after a lot of experiments) concluded was an optimal culture medium for citric acid production (note that this is *not* the same as the medium that optimizes growth of the fungus!). Not even going to try to summarize it; if someone's trying to start a company (Pfizer has been at this for about a hundred years... good luck competing with them!) they'll want to read the whole thing.

Sedit - 24-4-2011 at 16:36

Isn't Aspergillus Niger what may have caused the so called Pharaoh curse since it grew in abundence on the walls of King tuts tomb giving many of the members high fevers leading to there deaths....

I would take a little bit of care in trying to culture this just to be on the safe side.

Random - 24-4-2011 at 16:47

It is I think, I managed to grow it by making calcium citrate and then leaving the salt moist (which looked like yoghurt) in the room that had walls full of different molds. After a few days black mold started growing there, but I destroyed the culture.

bbartlog - 24-4-2011 at 17:22

People breathe in mold spores (including aspergillus) every day. Unless you're immunocompromised or have reason to think you're allergic to mold, I wouldn't take more than common sense precautions (gloves, maybe a dust mask).

Waffles SS - 24-4-2011 at 21:55

Thanks bbartlog.this book contain useful information.after reading this book i find out:Amount of Phosphate,Nitrate,Potassium and PH(provide with HCl) is very important to produce certain amount of Citric acid or Oxalic acid or Mycelium in 8 days.(I put this book here for another members.)
But i have some question:What size of plate or medium or baker(Erlen Meyer) is needed for produce 1kg Citric acid in one period(8 days)?Its funny but my friend decide to build a huge plastic pool for Citric acid fermentation but i think this is important that what depth of this pool should be full of reagent and fungi and also it should be complete sterile.
Excuse me but my information about microbiology is little and i am trying to improve it.

Attachment: 15.full.pdf (1.2MB)
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[Edited on 25-4-2011 by Waffles SS]

bbartlog - 25-4-2011 at 05:38

p23 of that Currie article has a chart showing grams of citric acid per 50ml of culture over 8 days. Notice that the results are sufficiently variable (0.3 to 3.5g) that it is difficult to provide a definite answer to your question; however if we are pessimistic and assume only 0.5g per 50ml then you would need a hundred liters of culture medium to produce a kilo of citric acid in eight days. But in this situation it would make sense to do some tests in a smaller volume to see how productive your particular culture+medium is, at which point you could answer the question yourself.
Anyway, if you have a pool I believe that you won't be able to increase production by filling it to more than a certain depth. Although the mycelium grows down in to the medium to some small depth (maybe 10cm? anyway I think it's in the article somewhere), diffusion doesn't proceed rapidly enough for greater depth to allow for further increases in production; basically you're limited by the surface area. The greater yields achieved in shallow pans (see p36, where 1 liter of medium produces ~80g of citric acid in 8 days) suggest that it's surface area rather than volume that should be considered when trying to project yields.

Kamikaza - 29-6-2011 at 07:48

A little bit offtopic, but would it be possible to make usefull amounts of oxalic acid by making a erlenmeyer with this growth medium:

1l water
3,0 gr NaNO3
0,5 gr KCl
0,5 gr MgSO4
0,1 gr FeSO4

and then putting in a little bit of "onion mold"? Are there any health hazards I should look out for, or are there efficienter ways to produce oxalic acid?

[Edited on 29-6-2011 by Kamikaza]

bbartlog - 29-6-2011 at 18:27

Your medium lacks phosphorus (also sugar but I'm assuming that's just an accidental omission). Further it's pretty useful to add a bit of HCl to keep other things from growing in it, though if you want to substitute impeccable sterile technique for a few ml of HCl, feel free. Anway, look at p22-23 of the abovementioned and linked book by Currie. Culture medium #3 looks like it maximizes oxalic acid production.

[Edited on 30-6-2011 by bbartlog]

Mildronate - 13-7-2011 at 09:53

In soviet time in my country was Citric acid manufacture, i had many equpment from there now. Few patents:

[Edited on 13-7-2011 by Mildronate]

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Mildronate - 13-7-2011 at 09:59

And more:

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Waffles SS - 5-3-2012 at 09:53

Quote:

Citric acid (CA) production by Aspergillus niger ATCC9642 from whey with different concentrations of
sucrose, glucose, fructose, galactose riboflavin, tricalcium phosphate and methanol in surface culture
process was studied. It was found that whey with 15% (w/v) sucrose with or without 1% methanol was
the most favourable medium producing the highest amount (106.5 g/l) of citric acid. Lower CA was
produced from whey with other concentrations of sugars and other additives used. Highest biomass of
A. niger was produced with the addition of riboflavins. In general, extension of the fermentation (up to
20 days) resulted in an increase in CA and biomass, and decrease in both residual sucrose and pH
values.

I just have access to aspergillus niger:ATCC 9142 ,ATCC 9029,ATCC 1004

Which one is more suitable for making citric acid?

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[Edited on 5-3-2012 by Waffles SS]

plante1999 - 5-3-2012 at 09:59

Quote: Originally posted by Waffles SS

Quote:

I just have access to aspergillus niger:ATCC 9142 ,ATCC 9029,ATCC 1004

Which one is more suitable for making citric acid?

[Edited on 5-3-2012 by Waffles SS]

Where can you buy these A. niger?

Waffles SS - 5-3-2012 at 10:04

My friend brought these A.niger for me.He bought it from iran
Which one is more suitable

GreenD - 5-3-2012 at 11:04

Why do so many people want citric acid? Just buy it... ?

Waffles SS - 5-3-2012 at 21:55

Quote:

Citric Acid Production by Aspergillus niger on the Ethanol Dry Milling Coproduct Thin Stillage

Seven strains of the fungus Aspergillus niger were screened for their ability to produce citric acid on the ethanol dry milling coproduct thin stillage. Citric acid and biomass production by the fungal strains grown on the thin stillage were analyzed using an enzyme assay and a gravimetric method, respectively. Citric acid production by A. niger ATCC 9029, ATCC 9142, ATCC 10577, ATCC 12846, ATCC 26550 and ATCC 201122 was similar after 144 h of growth on thin stillage. These strains also exhibited high citric acid specific productivities and yields. Only the citric acid production, specific productivity and yield by A. niger ATCC 11414 were lower after growth on thin stillage. Biomass production by ATCC 9029 on thin stillage was the highest of the strains studied. For all strains, at least 88% of the reducing sugar concentration in the thin stillage was consumed after 144 h
http://scialert.net/fulltext/?doi=jm.2007.678.683

Best result achived by ATCC 201122

I want to try whey with 15% (w/v) sucrose method.but really i dont know where sell whey!!(i think i should make it myself) Here i know where sell water of yogurt .i think this is same as whey(water of cheese)

bbartlog - 8-3-2012 at 10:36

Water of yogurt is similar to sour whey I imagine (sweet whey would be a different story). Should be OK for you, the aspergillus won't mind the lactic acid.

Waffles SS - 8-3-2012 at 23:17

Thanks.Sweet whey?what this mean?i found fresh whey and yogurt water.these are not Sour.
My whey should be Sour or sweet?
I read some where that said"you should separate spore by loops from PDA and i need 1x100000 spore per ml".What is loops?i cant pour my PDA content directly into my 4 Erlenmeyer?i can divide it into 4 portion?
I am sorry but my information is little about Biochemistry and i like to improve it.

[Edited on 9-3-2012 by Waffles SS]

bbartlog - 9-3-2012 at 05:28

Sweet whey is what you have if you make curds from milk using rennet. The remaining liquid is called sweet whey because it *is* sweet: it contains a bunch of lactose which has not yet been fermented. If the whey is left to ferment, the lactose is converted to lactic acid by lactobacillus at which point the whey is sour (and this is what would normally be considered whey).
Anyway, in the case of yogurt, fermentation has taken place and I would expect the yogurt water to be similar to sour whey for that reason.
I have no idea about your other question regarding loops.

Waffles SS - 9-3-2012 at 13:00

thanks bbartlog
Somebody know what means log 8/ml?

Quote:

A. niger ATCC 9642 stock culture was reactivated and cultivated
by streaking a loopfull of the culture on Petri dishes containing
solidified acidified(with 10% tartaric acid) potato dextrose agar
(PDA) and incubated at 250C for 5 days. Spores formed were
washed out twice with 10 ml distilled sterilized water each time.
Spore suspensions containing about log 8/ml were prepared and used as inoculums for the fermentation process.

Spore suspensions containing about log 8/ml were prepared
What this mean?(100.000.000 spore per ml?)

White Yeti - 10-3-2012 at 16:29

I tried making citric acid by this method. My first attempt failed and I didn't try again ever since. I am afraid of spores of this fungus, as I have heard some disturbing stories about A. niger. If I ever culture black mold again, it will be outside with a bottle of bleach close at hand

I will try it again this summer, or whenever it'll get warmer outside, so I can leave the petri dish outside, sheltered from wind of course.

I'll post results (if any) whenever I try this again.

Waffles SS - 10-3-2012 at 22:15

Why you afraid?Most of chemist think aspergillus niger is monster?its more dangerous than ebola?

White Yeti - 11-3-2012 at 08:00

Quote: Originally posted by Waffles SS

Why you afraid?Most of chemist think aspergillus niger is monster?its more dangerous than ebola?

In the summer, it gets extremely humid where I live. I remember one summer, it got up to around 80% relative humidity for almost a week. Fungal growth can be a real problem in my area and growing a petri dish full of this fungus indoors will not help out the situation.

As a matter of fact, fungal spores can be a major health hazard in normal people in high concentrations. Lungs provide a great place for fungus to grow, plenty of air, high humidity et-cetera.

I'd rather be safe than sorry. If you want to visualise how many spores come off of a mature sample of fungus when you lift the lid, you can go in a dark room with one window (which sheds a beam of light). Put your sample in the light, lift the lid and observe the cloud of spores that comes off. Bearing in mind that every one of those spores is an opportunistic pathogen, you might want to put the lid back on, go outside and breathe in some fresh air.

Waffles SS - 12-3-2012 at 08:59

Somebody know how many milliliters Spore suspensions(1x100000) is needed for 1 liter solution in 3 lit erlenmeyer?(my solution contain whey + 15% sucrose)

GreenD - 12-3-2012 at 09:08

can anyone explain why none of you will just buy citric acid?

White Yeti - 12-3-2012 at 16:21

GreenD anyone can buy citric acid. If I needed any for a serious synthesis I'd buy it. This thread is for making citric acid for the hell of it. If you think it's stupid, complain somewhere else.

Bot0nist - 12-3-2012 at 16:28

Quote: Originally posted by GreenD

can anyone explain why none of you will just buy citric acid?

For the same reasons I make my own nitrocellulose lacquer and other available things. I just like to make it. That is my hobby. For many of us on here, it is about the means and the learning adventure, about proving that we can do it, not about the end product.

Waffles SS - 12-3-2012 at 22:00

Quote: Originally posted by GreenD

can anyone explain why none of you will just buy citric acid?

Because : Chemistry =Chem+Is+Try and not buying!
(I saw this Sentence in this forum but i forgot where)

GreenD - 13-3-2012 at 06:34

Its just so many people were interest I thought I was missing something.
Thanks! I'm not complaining / bashing I'm just genuinely curious. If thats a problem go complain somewhere else!