We need an easy spectrophometric method to analysis amino acid of protein from yeast or other material. If you can recommend us alternative method without chromatographic way.
As I recall proteins which contain amino acids absorb UV at 280 nm. But this does not tell you what or how many amino acids are present. Alternatively, you can always acid digest the sample and perform a Kjeldahl Nitrogen (from the amide bonds).
What do you want to know, the total amount of amino acids in the protein, or the composition of amino acids? To measure the total amount of amino acids, you can hydrolyze the protein with heat and HCl, then react with an amine-derivatizing agent such as ninhydrin, and compare the color with a standard curve.
If you want to know the amino acid composition, a separation step is required, but it might be easier and cheaper to send the sample to a commercial or university analytical lab.
If you have access to it you can use ESI or MALDI mass spectrometry, they give you the protein fingerprint, so quite straightforward.
1. Kjeldahl digestion converts amino nitrogen into (NH4)2 SO4; Presence of ammonium salts in samples is disturbing.
2. Lowry's method is far more sensitive than Bradford's.
Literature:
Protein assay technical handbook , 2001 Pierce
Sapan, C.V., Lundblad, R.L and Price, N.C. (1999) Biotechnol. Appl. Biochem. 29, 99-108
Review Colorimetric protein assay techniques
Used it routinly those years.
Dear Marius,
Please check the following.
Regards
SM Najim
http://www.sgs.com/en/news/2016/06/amino-acid-analysis
https://www.hindawi.com/journals/jaa/2016/7374316/
https://www.hindawi.com/journals/jaa/2016/7374316/
http://pubs.rsc.org/en/content/articlelanding/2016/ay/c5ay02736e#!divAbstract
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