REFLECT AND APPLY An amino acid mixture consisting of lysine, leucine, and glutamic acid is to be separated by ion-exchange chromatography, using a cation-exchange resin at pH 3.5, with the eluting buffer at the same pH. Which of these amino acids will be eluted from the column first? Will any other treatment be needed to elute one of these amino acids from the column?

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Interpretation Introduction Interpretation: The amino acid among the given amino acids that will be eluted first from the column using a cation-exchange resin and eluting buffer at pH 3.5 is to be determined. Also, any other treatment that might be needed to elute one of these amino acids from the column is to be specified. Concept introduction: Ion-exchange chromatography is a technique that separates the substances on the basis of their charge. In this technique, an ion-exchange resin containing a ligand having either a positive or a negative charge is often used in the column. A positively charged resin serves as an anion exchanger, whereas a negatively charged resin acts as a cation exchanger. The pH at which a molecule or protein (amino acids) has no net charge is often termed as its isoelectric point. At a pH which is higher than the isoelectric point, the molecule acquires a net negative charge, whereas, at a pH that is lower than the isoelectric point, the molecule acquires a net positive charge. Explanation Lysine, leucine, and glutamic acid are the amino acids present in a mixture that is to be separated by ion-exchange chromatography, using a cation exchange resin and eluting buffer at pH 3.5 . In ion-exchange chromatography, a glutamic acid at pH 3.5 will have no net charge as pH of 3.5 is very close to its isoelectric point. As a result, the elution of glutamic acid will take place from the column in the ion-exchange chromatography. However, lysine and leucine will stick to the cation-exchange resin as both of the amino acids, at pH 3

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Biochemistry

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Mary K. Campbell + 2 others

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ISBN: 9781305961135

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Biochemistry

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Mary K. Campbell + 2 others

Publisher: Cengage Learning

ISBN: 9781305961135

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Interpretation Introduction

Interpretation:

The amino acid among the given amino acids that will be eluted first from the column using a cation-exchange resin and eluting buffer at pH 3.5 is to be determined. Also, any other treatment that might be needed to elute one of these amino acids from the column is to be specified.

Concept introduction:

Ion-exchange chromatography is a technique that separates the substances on the basis of their charge. In this technique, an ion-exchange resin containing a ligand having either a positive or a negative charge is often used in the column. A positively charged resin serves as an anion exchanger, whereas a negatively charged resin acts as a cation exchanger.

The pH at which a molecule or protein (amino acids) has no net charge is often termed as its isoelectric point. At a pH which is higher than the isoelectric point, the molecule acquires a net negative charge, whereas, at a pH that is lower than the isoelectric point, the molecule acquires a net positive charge.

Explanation of Solution

Lysine, leucine, and glutamic acid are the amino acids present in a mixture that is to be separated by ion-exchange chromatography, using a cation exchange resin and eluting buffer at pH 3.5. In ion-exchange chromatography, a glutamic acid at pH 3.5 will have no net charge as pH of 3.5 is very close to its isoelectric point. As a result, the elution of glutamic acid will take place from the column in the ion-exchange chromatography. However, lysine and leucine will stick to the cation-exchange resin as both of the amino acids, at pH 3

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