I am interested in this issue, regarding the checking of protein concentration in fermentation sample.
Dear Hiral,
My end product is lactic acid.
I run the simultaneous saccharification and fermentation (SSF) by using Rhizopus sp [Fungi] and cellulase enzyme + together with lignocellulosic substrate (carbon source) others mineral (Zn, Fe, K2HPO4, NH4SO4-amonia).
So, in this case still need to check protein?
Thank you.
If you are characterizing the amino acids in the fungal cell then as far as I know measurement of protein concentration is essential because deficiencies in these nutrients can reduce desired yields or alter final product quality. I hope this may be useful to you.
Protein concentration is checked to know the altered level of nitrogenous compounds in broth.
Protein concentration level is also important when extracellular protein or enzyme synthesis was expected
Dear All,
Thank for the sharing the knowledge.
I had performed the Bradford protein assay.
*Standard curve (BSA protein), 0.0-1.4 mg/ml or 0-1400 ug/ml.
The sample values read against standard curve were within range
0.02-0.13 mg/ml equivalent 20-130 ug/ml.
1. Is the value obtain too low? Need to be converted in ug/ml?
(FYI, I had heated the fermentation sample at 15 min at 95-100 C to denature the added enzymes, then cool down and keep in -20C, protein checking done after completion of fermentation cycle, 1-week)
2. Will this contribute to low protein value?
3. I need to redo the standard curve with lower BSA protein amount?
Thanks again.
it looks like your medium does not have the protein source, if it is so then measuring the protein content will give you estimate of the enzymes concentration that are released to break down the poly-sugars (in your case it is cellulose), if you add the cellulose enzyme into the medium then you can estimate the protein that belongs to this enzyme, be careful with enzyme addition because it might be used as a protein source breaking down by the released proteinases.
Protein concentration check is the necessary part because if you are working on an enzyme and you are chking it for maximal production then u have to optimize both the protein and activity simultaneously by calculating the specific activity of ur fermentation broth sample. It is needed coz if u have thinking of getting a purify product from that broth then u must go to do all purification steps which means there will be loss of proteins during ur purification processes. So to minimize heavy loss of protein you have to chk the first fermentation broth for higher specific activity so as at the end u will also get a good protein having good activity and potentiality. Hope I answered ur question. All d best.
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