Amino Acid Lab Report

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The amino acids are stained purple and from this we can measure relative distance traveled (RF) to determine which amino acids are present in each sample. The more hydrophobic the amino acid the the further up the paper it appears this is why leucine is above alanine. Alanine is more hydrophobic than glutamate which is at the bottom of the paper chromatography. The paper chromatography proved that only sample A had everything required for the GPT reaction to occur. This was indicated by the presence of both glutamate and alanine. As alanine wasn’t added to this sample it must have been produced by the reaction. This reaction contained the GPT enzyme in the homogenate solution and the substrates sodium L-glutamate and sodium L-pyruvate. The …show more content…

This is seen since there is no reaction between L-leucine and L-pyruvate to produce a second amino acid. If this had been a successful reaction it would be seen on the paper chromatography as two purple spots at different heights, one corresponding to the substrate L-leucine and one due to the product amino acid. The thin layer chromatography also would have had a second yellow spot if the reaction had proceeded as there would have been both the substrate and product -keto acid present. Reaction C substitutes sodium L-glutamate for sodium D-glutamate. This does not react to create alanine as seen on the paper chromatography therefore we can say that GPT is very specific since it can distinguish between these enantiomers. The L-glutamate is found animals but D-glutamate is not so the GPT enzyme has evolved to only react with L-glutamate. We can also see on the thin layer chromatography that no reaction occurred here as there is only the presence of the substrate -keto acid, pyruvate shown in yellow. Reaction D contains a phosphate buffer instead of an amino acid which gives no purple spots of the paper chromatography. This is because the there was no amino acids present in the substrate to react with the ninhydrin reagent and there was also no reaction to produce amino

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