July 07, 2020, 11:35:19 PM
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Topic: polymer and enzyme filtration  (Read 394 times)

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Offline NiccUnamebro

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polymer and enzyme filtration
« on: June 28, 2020, 09:02:06 AM »
Hello fellow chemistry lovers!

I had a few questions and was hoping you guys would have an answer. My first question is as follows: I am trying to calculate the amount of monomer units from cellulose. Lets say I have 250 grams of cellulose, how many monomer units would that be? I know that one cellulose monomer is (C6H10O5)n with μ = 162,1406 g/mol however I could not think of a way to calculate the amount of monomer units.

My second question was a way to filter out enzymes. Cellulase to be exact. At one point I have a substrate with cellulase enzymes in it, but I have to distillate the substrate which means I will lose my cellulase enzymes due to denaturation. Does anyone know a, relative 'easy', way to filter these enzymes out so that they could be re-used later on?

Thank you in advance!

Offline Borek

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Re: polymer and enzyme filtration
« Reply #1 on: June 28, 2020, 09:34:15 AM »
With known mass and molar mass you can calculate number of moles, no? And what is a mole?
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Offline Babcock_Hall

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Re: polymer and enzyme filtration
« Reply #2 on: June 28, 2020, 10:37:29 AM »
What substrate are you trying to distill?  There are many ways to separate an enzyme from its substrates, but it depends on the specifics of the situation.

Offline NiccUnamebro

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Re: polymer and enzyme filtration
« Reply #3 on: June 28, 2020, 11:07:29 AM »
With known mass and molar mass you can calculate number of moles, no? And what is a mole?

Oh, thank you! I realized that I can multiply the amount of moles with the constant of Avogadro and that would equal my monomer units right?

Offline NiccUnamebro

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Re: polymer and enzyme filtration
« Reply #4 on: June 28, 2020, 11:10:32 AM »
What substrate are you trying to distill?  There are many ways to separate an enzyme from its substrates, but it depends on the specifics of the situation.

Thanks for your reaction! The substrate mainly contains ethanol, the enzymes, citric acid, citrate, cellulose, glucose and brewers yeast.

Edit: The pH is about 5,0 and temperature is 37 Celsius (optimum for enzymes)

Offline Babcock_Hall

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Re: polymer and enzyme filtration
« Reply #5 on: June 28, 2020, 12:08:00 PM »
It is a Forum Rule that you must show your attempt to answer before we can help you.  What separation methods besides distillation do you know?

Offline Borek

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Re: polymer and enzyme filtration
« Reply #6 on: June 28, 2020, 01:57:22 PM »
I realized that I can multiply the amount of moles with the constant of Avogadro and that would equal my monomer units right?

About as accurately as possible.
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Offline NiccUnamebro

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Re: polymer and enzyme filtration
« Reply #7 on: June 28, 2020, 03:37:13 PM »
It is a Forum Rule that you must show your attempt to answer before we can help you.  What separation methods besides distillation do you know?

I absolutely understand! Well my thinking process before consulting here was as follows: First of all, separation based on difference on boiling point is a no go due to denaturation of the enzymes. Second of all (if possible) perhaps the usage of active carbon, but I would not know to which substance it will stick. My main goal is to filter out the ethanol and re-use the enzymes at some point. Other than these few separation methods, I have not really learned much methods at my school. I thought perhaps chromatography, but that is more qualitative and quantitative methods iirc. Then I went on google scholar to look for similar experiments (removing enzymes from a substance without damaging them), but I could not really find something that suited my purpose. I did come across something with a gel, electrophoresis to be exact, but I do not really have experience with it.

Offline Babcock_Hall

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Re: polymer and enzyme filtration
« Reply #8 on: June 28, 2020, 03:41:50 PM »
I should explain in advance that I am not sure how practical from an economic perspective it would be to recover both substrates and products and also recover enzymes in every situation, but doing is is certainly possible.

What do you know about the relative sizes of molecules such as ethanol and citrate versus the size of a typical enzyme? 

Offline NiccUnamebro

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Re: polymer and enzyme filtration
« Reply #9 on: June 28, 2020, 04:04:10 PM »
I should explain in advance that I am not sure how practical from an economic perspective it would be to recover both substrates and products and also recover enzymes in every situation, but doing is is certainly possible.

What do you know about the relative sizes of molecules such as ethanol and citrate versus the size of a typical enzyme?

I found that ethanol has a relative size of 0.44 nm and cellulase has a relative size of 4-6.5 nm. I could not find the relative size of citrate. So a way to filter the enzymes out is to just filter them out with filterpaper?

Offline Babcock_Hall

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Re: polymer and enzyme filtration
« Reply #10 on: June 28, 2020, 04:39:58 PM »
Ordinary filter paper would not work, although there is a technique called ultrafiltration that is another matter.  There is also a type of chromatography that goes by various names, one of which is gel filtration chromatography.  It is often used to perform what is sometimes called "buffer exchange" with respect to proteins.

Offline MNIO

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Re: polymer and enzyme filtration
« Reply #11 on: June 30, 2020, 04:00:43 AM »
If I understand your question, you are asking about how to separate cellulase enzyme from a solution/suspension of ethanol, citric acid, __ citrate, cellulose, glucose and brewers yeast (and probably water) to reuse it, and you are wondering if microfiltration will work.

In my professional opinion, not knowing the process conditions nor the slurry parameters but having spent a long time in this field, I'm going to say, no.  You won't have any luck doing this and should not waste your time chasing it.  There are too many variables that go into filtration design, your cellulase is too small to separate via microfiltration, microfiltration is ridiculously expensive, the process is a beast to operate, your yield would be terrible and last but not least, there's no guarantee any recovered cellulase would be active in your next batch of moonshine.  I don't think it can be done nor should be attempted.  The filter peddlers might disagree, but I wouldn't do it.

The way cellulase is recovered in industry is by crude filtration of the suspension to remove yeast, followed by precipitation with ammonium sulfate, followed by separation via adsorption (such as chromatography) columns. This paper discusses it.  I'm sure if you google you'll find more of the same info
http://scienceandnature.org/IJSN_Vol6(3)J2015/IJSN-VOL6(3)15-28.pdf
note the words "The sample containing maximum cellulase activity was
selected for further purification by chromatography" under the Chromatographic Technique discussion.  What do you read that as? I see it as a problematic purification process.

Anyway and more importantly, when you can buy it from Alibaba for $80/kg (or less in bulk), you need to ask yourself... "self, what business do I want to be in.  The business of making moonshine or the business of making cellulase".  If the later, you've got some learning to do, some redirecting of resources and some advanced marketing to work on, etc to be competitive in that market.  I wouldn't do that either.  I'd stick to making bourbon (or whatever it is you're making).

   

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