Hello Every one,
I have done saccharification with 50 U/ml crude enzyme extract in 50 ml buffer with 5% solid loading. Now I have a doubt at calculation part. If I get released reducing sugar of 1.23 g/ml what is my % of saccharification? I have taken 1 ml of crude enzyme extract for estimation of sugars released. Now I have to calculate it for 1 ml or 50 ml please let me know in detail in this regard. I am using the following formula:
Saccharificatioon (%) = sugars released x 0.9/ cellulose content x 100
Thank you,
Ramanjaneyulu, G.
Hi Golla
You can calculate it for "Unit of used enzyme" this is is a better value for scale-up.
Take care that you measure a blank of your reaction and of your enzyme, some lignocellulytic enzymes are coming from complex carbohydrate reactions and contain sugars itself. I dont know what the function this 0,9 is, but you can rely it directly to the amount of released sugar, per hour, for example.
Activity is always relative to your substrate, so if you want to check general activities, a purified, powdererd Cellulosis would be a good standard substrate; or you used coulored Substrates like Azo-Carbohydrate Substrates for a check.
pl find the attached article for your reference and calculations
See this publication:
https://www.researchgate.net/publication/281461543_Citric_waste_saccharification_under_different_chemical_treatments
This is a relation between sugars concentration per volume of licor divided by biomass used. If you want to calculate % saccharification based on carbohydrates present in the biomass, you need to multiply the biomass per %carbohydrates of biomass like 25%, you multiply per 0.25 the used biomass.
Article Citric waste saccharification under different chemical treatments
Saccharification is the process of producing monosaccharides from oligo- and poly-saccharides. Since polysaccharide of your interest is cellulose, saccharification % can be calculated from
Saccharificatioon (%) = glucose released x 0.9 x 100/ cellulose content
Glucose content should be measured either by GOD-POD method or glucose hexokinase method and not by DNSA method.
You have used 50U/ml of of enzyme in 50 ml buffer with 5% solid loading. You have released 1.23 g/ml reducing sugar. So total sugar in 50 ml will be 50 x 1.23.g. 5% Loading in 50 ml means 2.5g of biomass used. Calculate the amount of total sugar present in 2.5g of biomass with NREL Protocal and then use the formula.
% saccharification yield: total sugar released x100/total sugar in biomass
% saccharification is calculated on the basis of the total amount of glucan/xylan content in biomass (pretreated/native). To calculate the concentration of the polymeric sugars from the concentration of the corresponding monomeric sugars, using an anhydro correction of 0.88 (or 132/150) for C-5 sugars (xylose and arabinose) and a correction of 0.90 (or162/180) for C-6 sugars (glucose, galactose, and mannose).
Therefore use 0.9 factor is used for glucan to glucose conversion. Formula used is correct.
Tirath Raj
Hi,
This calculator on the web may help you.
https://www.alfa-chemistry.com/solution-dilution-calculator.htm
You might find the following paper to be of some help.
Article Enzymatic saccharification and fermentation of cellulosic da...
Dear everyone,
I would like to ask how to calculate the % xylose saccharification yie . I got 1 g/L of xylose in 5mL reaction with 2U of enzyme. I used 3% of substrate (corn hull) and suppose that 33 % of xylan content contain in corn hull. Please help me .
Thank you very much and be safety
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