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InvisibleAdoreChampignons
Mycophilic One

Registered: 08/10/08
Posts: 337
Loc: Seti Alpha 5
A Recipe For Homemade Soy Peptone?
    #9030251 - 10/04/08 09:51 PM (15 years, 8 months ago)

I've always liked the idea of supplementing agar mediums with peptone, but was always bothered by the price.  I've read in a couple places that soy peptone is made by digesting soy flour with papain.  Papain is readily available as a digestive aid in health food stores. I believe a bottle of papain tablets sells for under five bucks.

It sounds pretty simple. Apparently all one has to do is mix a tablet of papain in a beaker with water and soy four.  Stir it on a magnetic stirrer for a couple hours, and it's done.  Even though papain is a digestive enzyme, it shouldn't hurt the mushroom mycelium. Papain begins to decompose at 85 degree Celsius.  It'll definitely loose its enzymatic activity once the medium has been pressure cooked.

Has anyone tried this before?  Is it that easy or did I miss something?


--------------------
There is no such thing as a dumb question.  There are just curious people trying to learn something new. 

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Offlinedenger
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Registered: 09/19/08
Posts: 1,177
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Re: A Recipe For Homemade Soy Peptone? [Re: AdoreChampignons]
    #9030301 - 10/04/08 10:10 PM (15 years, 8 months ago)

What you will get with papain is not exactly the same as classical peptone. According to wikipedia, peptones are derived from animal milk or meat digested by proteolytic digestion. In addition to containing small peptides, the resulting spray-dried material includes fats, metals, salts, vitamins and many other biological compounds.
I need to add that frequently, it is an acid-induced (most often HCl) proteolysis instead of enzymatic, because its cheaper that way.
If you taste your expensive peptone, and it tastes salty, its HCl-derived. If it tastes like unsalted chicken broth, then its pepsin or trypsin derived.
All of the above may not matter for the purpose of making media, but your home made concoction will not be 100% digested in all likelihood, and it will be very hard to come up with a dosage equivalent to regular peptone.
Besides, for about $30 you can buy enough for 20 liters of media here:
http://www.bio-world.com/MoreInfo.php?ItemCode=760092

How much do you really need?

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InvisibleAdoreChampignons
Mycophilic One

Registered: 08/10/08
Posts: 337
Loc: Seti Alpha 5
Re: A Recipe For Homemade Soy Peptone? [Re: denger]
    #9030613 - 10/04/08 11:40 PM (15 years, 8 months ago)

Quote:

denger said:
What you will get with papain is not exactly the same as classical peptone. According to wikipedia, peptones are derived from animal milk or meat digested by proteolytic digestion. In addition to containing small peptides, the resulting spray-dried material includes fats, metals, salts, vitamins and many other biological compounds.
I need to add that frequently, it is an acid-induced (most often HCl) proteolysis instead of enzymatic, because its cheaper that way.
If you taste your expensive peptone, and it tastes salty, its HCl-derived. If it tastes like unsalted chicken broth, then its pepsin or trypsin derived.
All of the above may not matter for the purpose of making media, but your home made concoction will not be 100% digested in all likelihood, and it will be very hard to come up with a dosage equivalent to regular peptone.
Besides, for about $30 you can buy enough for 20 liters of media here:
http://www.bio-world.com/MoreInfo.php?ItemCode=760092

How much do you really need?




Greetings Denger,
yes, I hear what you're saying.  For myself though, I tend to learn from the ground up.  Instead of telling myself I need to buy this from there or that from here, I'd like to see how far I can go in synthesizing compounds that I need from scratch.  It's not that I'll always follow through with what I discover, it's just that I'd like to know the whole process.  For example in reading up on the laboratory preparation of papain, I discovered that one collects the raw papain from the sticky sap of the papaya plant, not from the fruit.  It's the sticky sap, or papaya plant latex, that is harvested.  It's then separated and purified in a solution of polyethylene glycol and ammonium sulfate.  While you won't see me trying to grow a papaya plant from seed in my backyard, it's learning about these fine details that fascinates me, propels me, and perpetuates my interest.

You're right though about the commercial grade soy peptone.  It may be more throughly digested.  Papain is a very selective protease.  It'll only cleave proteins at certain junctures leaving the rest untouched.  If I wanted soy protein completely broken down to its constituent amino acids, hydrochloric acid, or HCl, could be used.  It is also a powerful non-selective protease.  Once digested, it could be neutralized with sodium hydroxide, NaOH.  It's the combination of using hydrochloric acid and sodium hydroxide that gives the peptone its salty taste.  Both hydrochloric acid and sodium hydroxide combine and neutralize to produce regular table salt, NaCl.

If I wanted to produce powdered soy peptone, I believe that adding in tannic acid and alcohol will encourage the hydrolyzed amino acids to precipitate out of solution making it possible to filter and dry it.

In general, peptone can come from variety of plant and animal sources.  However, in knowing that we're trying to grow saprophytic fungi, it makes sense that fungal mycelium would prefer amino acids digested from plant protein sources, or soy peptone.  If I were growing mammalian pathogenic bacteria, it would make sense that it would prefer amino acids broken down from animal protein sources.

I hope I didn't drive you batty with my length reply.  It just fascinates me to know how everything comes together to produce the final product.


--------------------
There is no such thing as a dumb question.  There are just curious people trying to learn something new. 

Edited by AdoreChampignons (10/05/08 12:07 AM)

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InvisibleCheeWiz
Male

Registered: 09/28/08
Posts: 276
Re: A Recipe For Homemade Soy Peptone? [Re: AdoreChampignons]
    #9033587 - 10/05/08 05:33 PM (15 years, 8 months ago)

  Neo Peptone & Peptone S are soy based peptones for Defco and others that have been around since the 60's if not longer.  This is a short course on how they are manufactured. Using a soy protein powder suspension in deionized water and treating it with Hydrochloric acid to a PH of about 4.7. This starts the digestion of the soy protein particles that are in the suspension. This digest is then treated with various peptic enzymes to farther digest the soy proteins. The end product has its PH adjusted and is then spay dried and ground to make a dry powder. A little too much work for me to go though.
  Peptones are very hygroscopic and can go a little off before you can use up a 100 gram bottle when using only two grams per liter (that's 50 liters of media). So I have stopped using it all to gather. What I do now is buy premixed media powder that I just need to add deionized water to and sterilize. These can be attained from our fine sponsors!

I need to up date the availability of peptic enzymes. I went on line to day (10/15/08) to reorder some more peptic enzyme from my brewing & wine making supplier to find that they no longer carry it, the same was true at the other suppliers on line. Peptic enzymes was sold to help clarify wine by reducing protein haze in it. Peptic and pectic enzymes are not the same, the later is still being sold to help brake down the pectin in fresh fruit pulp so one can extract the most juice out of fruit when making fruit wines.

  But as of late I have been making an oat flake agar media in which I use barley malt flour that has been cold processed with most of its diastase intact and peptic enzymes. The use of malt flour coverts some the starches to simple sugars and dextrin, while the peptic enzymes helps digest the proteins into simple free proteins. Heating to a boil will denature these enzymes. The formulation is quite simple and is as follows:
60 grams of organic oat flakes
5 grams barley malt flour
1 gram organic soy flour
1 gram nutritional yeast as a flake or powder
Peptic enzymes, use per manufactures recommendations and adjusted for a liter.
  Into 500 ml of room temperature di water mix all of the above, letting rest over night and then heat to a simmer for few minutes. Remove from heat and filter though a fine filter adding enough di water to make a liter. Slowly reheat while adding 25 grams agar tell the agar dissolves and remove from the heat, add any buffers, trace elements, vitamins and antibiotics you are going to use and process as with any other agar.
  This agar does have a somewhat of a soft set. But I have found it very useful and the only one I now use. It has a good balance of simple sugars, dextrin, complex carbohydrates, soluble fiber, lipids, simple free proteins and vitamins.
  The barley malt floor, organic oat flakes, organic soy flour, agar and nutritional yeast can be found at most health or whole food store‚Äôs and the peptic enzymes can be found at wine making shops or all can be found on line.

Edited by CheeWiz (10/16/08 12:42 PM)

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OfflineAcidHorse
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Re: A Recipe For Homemade Soy Peptone? [Re: AdoreChampignons]
    #9033899 - 10/05/08 06:30 PM (15 years, 8 months ago)

8156 PET TABS SUPPLMNT 60TB -- 8156
8156 PET TABS SUPPLMNT 60TB
8156
Product Use: A HIGHLY PALATABLE VITAMIN-MINERAL SUPPLEMENT FOR DOGS.

Label Instructions: PUPPIES AND DOGS UNDER 10 LB -1/2 TABLET DAILY. DOGS OVER 10 LB -1 TABLET DAILY.

Ingredients: WHEAT GERM KAOLIN CORN SYRUP PORK LIVER MEAL DICALCIUM PHOSPHATE SAFFLOWER OIL SUCROSE LACTOSE GELATIN CORN STARCH STEARIC ACID NIACINAMIDE HYDROLIZED VEGETABLE PROTEIN IRON OXIDE AND PEPTONE MAGNESIUM STEARATE AND OTHERS...

Manufacturer: VIRBAC CORP.

http://www.1stqualitypetsupplies.com/pet-supplies-dog-grooming-aids-health-aids-nutrition-305400.html


--------------------
If you wanna ride, don't ride the white horse, if you wanna ride, ride the white pony

Edited by AcidHorse (10/05/08 06:31 PM)

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Offlinedenger
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Re: A Recipe For Homemade Soy Peptone? [Re: AcidHorse]
    #9034053 - 10/05/08 07:07 PM (15 years, 8 months ago)

Peptone and Magnesium stearate are two different unrelated things.
The fact that peptone comes right before binder, it probably means they contain very darn little of it. I wont use these tabs.

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OfflineAcidHorse
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Re: A Recipe For Homemade Soy Peptone? [Re: denger]
    #9034173 - 10/05/08 07:36 PM (15 years, 8 months ago)

Quote:

denger said:
Peptone and Magnesium stearate are two different unrelated things.
The fact that peptone comes right before binder, it probably means they contain very darn little of it. I wont use these tabs.




No I'm not pointing out to use theses, there may be supplements out there that would have this protein say in a formula for infant animals.

And there would have to be a whole lot in a formula for infants otherwise they would die due to lack of it.


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If you wanna ride, don't ride the white horse, if you wanna ride, ride the white pony

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OfflineAcidHorse
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Re: A Recipe For Homemade Soy Peptone? [Re: AdoreChampignons]
    #9034186 - 10/05/08 07:39 PM (15 years, 8 months ago)

Actually I think AdoreChampignons is looking for Casein Peptones
http://www.organotechnie.com/caseinpe.htm

As per Volk's PDF document on morel sclerotia.


--------------------
If you wanna ride, don't ride the white horse, if you wanna ride, ride the white pony

Edited by AcidHorse (10/05/08 07:40 PM)

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Invisiblelipa


Registered: 07/24/07
Posts: 2,684
Trusted Cultivator
Re: A Recipe For Homemade Soy Peptone? [Re: CheeWiz]
    #9035216 - 10/05/08 11:34 PM (15 years, 8 months ago)

Hipster
Thanks for the agar recipe. I am going to try it. Sounds good!

LIPA

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OfflineAeolus1369
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Registered: 05/20/02
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Re: A Recipe For Homemade Soy Peptone? [Re: AdoreChampignons]
    #9074367 - 10/14/08 12:56 AM (15 years, 8 months ago)

Quote:

AdoreChampignons said:
It sounds pretty simple. Apparently all one has to do is mix a tablet of papain in a beaker with water and soy four.  Stir it on a magnetic stirrer for a couple hours, and it's done.  Even though papain is a digestive enzyme, it shouldn't hurt the mushroom mycelium. Papain begins to decompose at 85 degree Celsius.  It'll definitely loose its enzymatic activity once the medium has been pressure cooked.

Has anyone tried this before?  Is it that easy or did I miss something?




I'm thinking if this enzyme is supposed to work in your stomach (e.g. acidic, with lots of other ions and stuff present) it will definitely not work in plain water.  The acidity and ionic strength of a solution are HUGE determinants of enzymatic stability and activity

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