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The elusive chocolate colored Aspergillus Niger var. Awamori

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The elusive chocolate colored Aspergillus Niger var. Awamori

Postby pintoshine » Thu Mar 05, 2009 9:16 pm

I've been playing again. I am not quite ready to present the entire project but some of my preliminary results are looking great.
I isolated another variety of Aspergillus Niger. This one is a chocolate colored variety.

This is the original capture.
Image

This is an isolated pin prick of spores transferred to my proprietary growth media.(others who have paid attention know my recipe).
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After 24 hours, this is the amount of growth it obtained. The diameter of the growth that is visible on top is 7mm.
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After 36 hours, this is the amount of growth it obtained. The diameter of the growth that is visible on top is 20mm.
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After 48 hours, this is the amount of growth it obtained. The diameter of the growth that is visible on top is the width of the cup about 50mm.
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You will notice the distinct ball shaped heads on the top of filament thin stalks. They look as if they are suspended in mid air.
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I added this information because it was really cool.

I did a pH check of the growth media because one of the criteria this has to pass is that it acidifies its host. The ph for this media was at 6 to 6.5. I expected this because of the nutrient content would have decreased the pH slightly.
Image

Another check that I make is the this mold does not discolor its growth media. After 84 hours growth the whole top of the media was covered with spores from the mature mold. I added a small amount of corn meal to use as an erosive, abrasive material to knock off and combine with the dusty mold spores. I then put the sample of corn meal away to use this batch of spores to create enough to distribute to my friends. The gelled media is liquid because I microwaved it. The A niger isn't producing enzymes to break down agar agar. Agar agar is extracted from sea weed and algae. It is a carbohydrate but it is long chains of galactose. Need I say that is what beano was created for. Galactose is a hard sugar to break down. Here is a shot of the media and the bottom of the solid, water tight membrane created by the mold. I say water tight because the liquid media would not come out of the cup until I broke the membrane. This is 84 hour of growth.
Image

This is the cup of media with the mold removed and discarded. I added a few drops of broad range indicator. It turned quit red at first and then went to a more dull red. The red indicated the presence of lots of acid. The pH was lower than the 4.0 the bottle of indicator scale goes to.
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Ok so here is the final two test before I make a big batch. What does it taste like? Oh I can hear the :shock: :shock: :shock: OOOOO's already. :screwy: :screwy:

The media itself contains no chemicals. It is all food grade. The media was heated to liquefy the agar agar. This heating mostly destroys most enzymes except for the amyloglucocidase which is very heat tolerant :idea: .

The original media tasted something similar to very weak semolina with a small amount of sugar. This is what we in the USA call Cream of Wheat. My mother always prepared ours with water and sugar.

The acidified gel tastes like .....

FRUIT!! :!: :!: Mission accomplished.

One more test which I did not photograph. The detractor are saying "Exposure to toxic Black mold will kill you."
So I tested it for aflatoxin, the only known bad toxin from the Aspergillus flavus strain. The test is simple and conclusive and my camer would not do anything but flare. I test the media for aflatoxin using the industry standard 365nm long UV light source. Ah, that is why the camera flared.
here is a link to an articale that explains the detection of aflatoxin.
http://www.pubmedcentral.nih.gov/picren ... obtype=pdf

The A. niger showed nothing but the blue illumination of the light and did not show any green luminance which indicates aflatoxin.

There is one more test to do and that is to do a real fermentation using the mold as a form of "Malt rice".

I am hoping to get this one developed for commercial sales for ones wanting to make Awamori. I have not been able to source a supply of this mold, so I developed my own. It should be up on the site around March 20, 2009 or so.

ps. for those of you that I have already promised a sample of A. niger, I'll be sending this one along with a small amount of A. oryzae (sake tane-koji) this brown one is called Kuro-koji. I also got hold of a native A. oryzae while I was hunting mold this round.
Enjoy the pictures.
Distill safe. Make good cuts. Enjoy yourself. Give as much as you take. Have fun doing this incredibly hard work. Be a good example. It's your hobby.
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Re: The elusive chocolate colored Aspergillus Niger var. Awamori

Postby Rebel_Yell » Thu Mar 05, 2009 10:47 pm

That is cool as two hells.
What are you using as an incubator?
Your results may vary..
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Re: The elusive chocolate colored Aspergillus Niger var. Awamori

Postby pintoshine » Sat Mar 07, 2009 10:44 am

I added content and pictures to the first post.
Distill safe. Make good cuts. Enjoy yourself. Give as much as you take. Have fun doing this incredibly hard work. Be a good example. It's your hobby.
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Re: The elusive chocolate colored Aspergillus Niger var. Awamori

Postby Rebel_Yell » Sat Mar 07, 2009 1:24 pm

:8) My hat is off to you Sir.
I have a son in college that I am suggesting that micro biology might be a good field to get into...
Your results may vary..
I would rather teach a pig to sing than argue with an Idiot.
Every sixty seconds you spend angry, upset or mad,
is a full minute of happiness you'll never get back.
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Re: The elusive chocolate colored Aspergillus Niger var. Awamori

Postby zymos » Sat Mar 07, 2009 5:16 pm

Nice work (and pix) :8)
I'd live to be able to get some of these cultures...
Hola Amigos!
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Re: The elusive chocolate colored Aspergillus Niger var. Awamori

Postby manu de hanoi » Sat Mar 07, 2009 11:19 pm

Rebel_Yell wrote::8) My hat is off to you Sir.
I have a son in college that I am suggesting that micro biology might be a good field to get into...

I did 3 years of biology and microbio from 95 to 98. Problem is it's the science of the future not the present, and that means it's very difficult to get a job in that field even after 8 years of study + n years of post doc, at least in europe. Tell him that too. However I'd be glad to work on that field as soon as I can. And if I can't, i'll buy myself a microscope and an incubator (and who knows perhaps some dna replication device!) as soon as I have the space and money. By the way, congrats pinto, that's some very nice research you done here.
Perhaps a drop of ethanol in the medium would help select etoh resistant strains.
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Re: The elusive chocolate colored Aspergillus Niger var. Awamori

Postby pintoshine » Sun Mar 08, 2009 10:55 am

Manu, I have seen a lot of research related to using this to ferment manioc(yuca/cassava). I have not been down that path yet but it is a checkbox.
I'm about half way done with the manual but haven't started on the editing yet. I have been learning advanced css to get the style I need for this presentation and manual. As soon as I get it done I'll be needing a couple proof readers.
Distill safe. Make good cuts. Enjoy yourself. Give as much as you take. Have fun doing this incredibly hard work. Be a good example. It's your hobby.
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Re: The elusive chocolate colored Aspergillus Niger var. Awamori

Postby Rebel_Yell » Sun Mar 08, 2009 12:22 pm

I would be happy to proofread it for you Pint.

I really have no idea what direction my youngster will decide to educate himself Manu. He has no idea what he wantes to be when he grows up...
Your results may vary..
I would rather teach a pig to sing than argue with an Idiot.
Every sixty seconds you spend angry, upset or mad,
is a full minute of happiness you'll never get back.
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Re: The elusive chocolate colored Aspergillus Niger var. Awamori

Postby manu de hanoi » Sun Mar 08, 2009 12:50 pm

pintoshine wrote:Manu, I have seen a lot of research related to using this to ferment manioc(yuca/cassava). I have not been down that path yet but it is a checkbox.
I'm about half way done with the manual but haven't started on the editing yet. I have been learning advanced css to get the style I need for this presentation and manual. As soon as I get it done I'll be needing a couple proof readers.

manual about what? if that's about the aspergillus i'd sure be glad to read.
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Re: The elusive chocolate colored Aspergillus Niger var. Awamori

Postby mtnwalker » Sun Mar 08, 2009 11:25 pm

Wow, Pint.

I have been trying to ascertain a simular mold like that for over 2 years. You sure got my interest up. The kojis as well.

Should get a doctorate for this. Young Frankenstein??
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Re: The elusive chocolate colored Aspergillus Niger var. Awamori

Postby pintoshine » Mon Mar 09, 2009 10:36 am

I need to say thank you to those who have offered kind words.
I need to answer some questions.

Q: What manual?
A: I am writing a manual on how to do this process from beginning to end. I am also adding chapters on the application of the enzymes. This will include using the "Malt" to convert rice, corn, cassava(yuca), white and sweet potato starches. All these are well suited to simi-solidstate fermentation due to the high water content after gelatinization.

Q: Is the process to grow this stuff hard?
A: Anyone who can do basic cooking can handle this stuff. It is almost as easy as pitching dry yeast in a fermenter.

Q: How can I get some?
A: I am beginning to farm this in quantity today, March 9,2009. It takes 6 days to get a very mature batch of spores. If you PM me your mailing address I'll send you some 1st class along with basic instructions.

Q: What will you send?
A: Currently I have a commercially sourced A. oryzae, a home isolated A. oryzae and the A. niger in the pictures above.

Q: Do you need special equipment such as an incubater?
A: No, you can do this any where it stays around 72F(22C). If you are in a colder location, then using a heated 2 liter bottle of water in a cooler will work fine to provide enough heat.

Q: Does it grow in the dark?
A: Yes, but I have much better luck with spores in alternating light like the natural day/night cycle.

Q: How long does it take to make "Malted Rice"?
A: If you have 72F fairly constant, after the rice is room temperature it is inoculated, well mixed every 12 hours or so, it will be 36 to 40 hours. Any longer and spores start showing. The time has to be adjusted to the temperature. Warmer is faster and cooler is slower. The optimal time is at the first sight of spore coloring. If you can catch it a few minutes, yes minutes, before spore formation it is the most enzyme rich.

Q: After I get a sample, can I grow more?
A: Yes, the farming of aditional cultures is very easy and quite fun to watch, because it progresses so fast. If you need to take a break, refrigeration of a spore coverd growing media container will last about 6 months to a year.


I hope this clears up some questions.
Distill safe. Make good cuts. Enjoy yourself. Give as much as you take. Have fun doing this incredibly hard work. Be a good example. It's your hobby.
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Re: The elusive chocolate colored Aspergillus Niger var. Awamori

Postby Rebel_Yell » Tue Mar 10, 2009 10:00 pm

Your answers clear lots of questions and start the itch...
Your results may vary..
I would rather teach a pig to sing than argue with an Idiot.
Every sixty seconds you spend angry, upset or mad,
is a full minute of happiness you'll never get back.
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Re: The elusive chocolate colored Aspergillus Niger var. Awamori

Postby Rebel_Yell » Tue Sep 21, 2010 4:45 pm

Good find Manu.
Your results may vary..
I would rather teach a pig to sing than argue with an Idiot.
Every sixty seconds you spend angry, upset or mad,
is a full minute of happiness you'll never get back.
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Re: The elusive chocolate colored Aspergillus Niger var. Awamori

Postby vb » Tue Sep 21, 2010 6:10 pm

thats some good info there.
See You Next Tuesday.
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