Site-selective deuteration of amino acids through dual protein catalysis
AssigneeWisconsin Alumni Research Foundation
InventorsAndrew Buller, Tyler Doyon
Life of this patent
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Abstract
A method for site-selective deuteration of amino acids using a protein system having an aminotransferase (e.g., DsaD) and/or a small partner protein (e.g., DsaE). A non-deuterated amino acid is contacted with deuterium and an aminotransferase or a combination of an aminotransferase and a partner protein, to yield a Cα-deuterated or a Cα- and Cβ-deuterated amino acid. Cβ-deuterated amino acids can be accessed by contacting a Cα- and Cβ-deuterated amino acid with non-deuterium hydrogen and an aminotransferase to wash out the deuterium at the Cα carbon atom by the non-deuterium hydrogen.
Claims13
A method of making a deuterated amino acid, the method comprising contacting a non-deuterated amino acid having an α-position carbon atom and a β-position carbon atom with deuterium and a protein selected from the group consisting of a branched-chain aminotransferase and a combination of a branched-chain aminotransferase and a partner protein, for a time and at a temperature to selectively deuterate the non-deuterated amino acid at the α-position carbon atom to yield a Cα-deuterated amino acid or to selectively deuterate the non-deuterated amino acid at both the α-position and the β-position carbon atom to yield a Cα- and Cβ-deuterated amino acid, wherein the branched-chain aminotransferase is a protein comprising an amino acid sequence at least 85% identical to SEQ ID NO:
- 1, and wherein the partner protein is a protein comprising an amino acid sequence at least 85% identical to SEQ ID NO:
- 3 or SEQ ID NO:
- 4.
A method of making a deuterated amino acid, the method comprising contacting a non-deuterated amino acid having an α-position carbon atom and a β-position carbon atom with deuterium and a combination of a branched-chain aminotransferase and a partner protein, for a time and at a temperature to selectively deuterate the non-deuterated amino acid at both the α-position and the β-position carbon atom to yield a Cα- and Cβ-deuterated amino acid, wherein the branched-chain aminotransferase is a protein comprising an amino acid sequence at least 85% identical to SEQ ID NO:
- 2, and wherein the partner protein is a protein comprising an amino acid sequence at least 85% identical to SEQ ID NO:
- 3 or SEQ ID NO:
- 4.
A method of making a deuterated amino acid, the method comprising contacting a non-deuterated amino acid having an α-position carbon atom and a β-position carbon atom with deuterium and a branched-chain aminotransferase, for a time and at a temperature to selectively deuterate the non-deuterated amino acid at the α-position carbon atom to yield a Cα-deuterated amino acid, wherein the branched-chain aminotransferase is a protein comprising an amino acid sequence at least 85% identical to SEQ ID NO:
- 1.
A method of making a deuterated amino acid, the method comprising:
- contacting a non-deuterated amino acid having an α-position carbon atom and a β-position carbon atom with deuterium and a combination of a branched-chain aminotransferase and a partner protein, for a time and at a temperature to selectively deuterate the non-deuterated amino acid at both the α-position and the β-position carbon atom to yield a Cα- and Cβ-deuterated amino acid;
- and
- contacting the Cα- and Cβ-deuterated amino acid with non-deuterium hydrogen and the branched-chain aminotransferase, for a time and at a temperature to selectively replace the deuterium at the α-position carbon atom with the non-deuterium hydrogen to yield a Cβ-deuterated amino acid;
- wherein the branched-chain aminotransferase is a protein comprising an amino acid sequence at least 85% identical to SEQ ID NO:
- 1;
- and
- wherein the partner protein is a protein comprising an amino acid sequence at least 85% identical to SEQ ID NO:
- 3 or SEQ ID NO:
- 4.
Dependencies are inferred from the claim text.