Re: [DIYbio] Making peptone

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I'm assuming the 600C is a formatting error for 60C? :)
I don't think Papain, or the experimenter, would survive kitchen
chemistry involving fluids at 600C..

Interesting that Papain (relative of bromelain, fairly interchangeable)
is so thermostable; I'd seen mention that that enzyme family can last at
60C for prolonged periods and that it's their optimum temperature.

I wonder; do plasma-associated trypsin inhibitors inhibit bromelain and
papain, given that they use the same three-amino catalytic site and
overall activity? That is, could one use bromelain/papain as substitutes
in mammal cell culture, where inactivation is achieved by addition of
calf serum (hopefully soon to be artificial)?

On 28/04/14 09:58, Xabier Vázquez Campos wrote:
> Here you even have a recipe to get peptone from raw soya beans.
> http://www.ajol.info/index.php/ajb/article/download/55823/44286
> If you have the protein extract isolated just follow from there:
>
> The pH of the protein slurry was adjusted to pH 6.5 with 1 N NaOH. At
>> optimum temperature of the digest (600C) 0.25% of commercial papain was
>> added. Digestion was allowed to take place for 3 h and the pH was
>> controlled using 1 N NaOH. The temperature was then raised to 800C for 5
>> min to inactivate the enzyme. Liquor from the hydrolysed soyabeans was
>> separated from the protein/enzyme slurry by repeated centrifugation. The
>> peptone solution was then freeze-dried to obtain soya peptone.
>>
>
>
> El lunes, 28 de abril de 2014 18:11:15 UTC+10, Cathal Garvey escribió:
>>
>> That may have been mine, I used the word "homebrew" at some point. :)
>>
>> The original document was early work, it was overly wordy and overly
>> careful. In the time since, I found that you don't have to be so
>> precise. These days the protocol is condensed, you'll find it as a
>> subheading in the E.coli transformation protocol;
>>
>> https://github.com/cathalgarvey/biohacking-protocols/blob/master/E.coli%20Transformation%20With%20PEG%2BMgSO4.md
>>
>> On 27/04/14 21:56, Avery louie wrote:
>>> IIRC it used meat tenderizer or something to break down the soy or
>> casein.
>>>
>>> if you want to do diy media, you might want to try potato dextrose agar,
>> as
>>> it is very simple to make.
>>>
>>> --A
>>>
>>>
>>> On Tue, Apr 22, 2014 at 3:13 PM, Joffrey <biok...@gmail.com<javascript:>>
>> wrote:
>>>
>>>> Hello,
>>>> i was reading an article on openwetware wiki, and in this article there
>>>> was a link to a recipe to make your own peptone from soy or casein but
>> the
>>>> links are dead or ask for password, does someone have these documents ?
>>>> here is the article in question
>>>> http://openwetware.org/wiki/DIYbio/FAQ/Methods the link to the
>> documents
>>>> for making peptone are at the end of the article under "Homebrew Growth
>>>> Media"
>>>>
>>>> many thanks
>>>>
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>> --
>> T: @onetruecathal, @IndieBBDNA
>> P: +353876363185
>> W: http://indiebiotech.com
>>
>

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