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Producing Partially Deuterated Amino Acids Using Aminotransferase and Amino Acid Oxidase Enzymes

IP.com Disclosure Number: IPCOM000075909D
Original Publication Date: 1971-Dec-01
Included in the Prior Art Database: 2005-Feb-24
Document File: 3 page(s) / 37K

Publishing Venue

IBM

Related People

Studebaker, JF: AUTHOR

Abstract

In the structural analysis of small protein hormones and antibiotics, one of the techniques employed has been nuclear magnetic resonance. In this technique, the spectral lines of the alpha hydrogens of these proteins are central to their structural determination. However, the application of nuclear magnetic resonance to the analysis of larger protein molecules has not been effective, because the spectral lines of the alpha hydrogens tend to increasingly overlap as more amino acids occur in the protein molecule chain.

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Producing Partially Deuterated Amino Acids Using Aminotransferase and Amino Acid Oxidase Enzymes

In the structural analysis of small protein hormones and antibiotics, one of the techniques employed has been nuclear magnetic resonance. In this technique, the spectral lines of the alpha hydrogens of these proteins are central to their structural determination. However, the application of nuclear magnetic resonance to the analysis of larger protein molecules has not been effective, because the spectral lines of the alpha hydrogens tend to increasingly overlap as more amino acids occur in the protein molecule chain.

Disclosed is a method wherein the above-stated deficiency is substantially overcome. The method essentially is the stereospecific production of amino acids which are deuterated at the alpha position using aminotransferase and amino acid oxidase enzymes. It is known that deuterated amino acids lend themselves advantageously to nuclear magnetic resonance studies of protein molecule structures. This method is simple and less expensive than methods now in use for the same purpose.

Heretofore, deuterated amino acids, isolated from algae grown in D(2)O, have been used for some of the residues in the protein chain. Since these deuterated amino acids do not contribute lines to the NMR spectrum, the latter is considerably simplified. However, such algae grown deuterated amino acids are very expensive to produce since there has to be provided in their production, the expensive D(2)O for all of the substances synthesized by the algae. Thereafter, the amino acids have to be first isolated from the algae and then from each other. As an example of current costs, lyophilized cells of algae grown in D(2)O cost about $5.00 to $6.00 per gram.

In accordance with this method, there are employed aminotransferase enzymes to catalyze the following reactions:

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In the above reaction, the hydrogen atom which is introduced at the alpha position of the keto acid, upon reaction, is provided by the aqueous solvent wherein the reaction takes place. According to the method, the solvent which is employed is D(2)O whereby a deuterium atom is introduced at the a position of the amino acid. Since the reaction is reversible, both of the amino acids therein would be deuterated provided that the reaction were permitted to proceed for a period equal to several reaction lifetimes. The amino and keto acids are readily separated, such as in a column of the Dowex...