Products of manufacture and processes for peptide synthesis
AssigneeDiversa Corporation
InventorsGrace Desantis, Mark J. Burk
Life of this patent
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Abstract
In one aspect, the invention provides a nontemplate directed, enzymatic thermo-cycled (NTDET) peptide synthesis process. The invention also provides products of manufacture comprising a reaction chamber for synthesizing a peptide or a polypeptide.
Claims73
An enzymatic process for synthesizing a peptide comprising the following steps
- (a) providing at least two amino acids, at least one peptide and at least one amino acid, or at least two peptides;
- (b) providing a peptide ligase and a deacetylase, wherein the peptide ligase and the deacetylase are active under different reaction conditions and the peptide ligase is active after exposure to reaction conditions where the deacetylase is active and the deacetylase is active after exposure to reaction conditions where the peptide ligase is active;
- (c) contacting the amino acids of step (a), or the peptide and the amino acid of step (a), or the peptides of step (a), with the peptide ligase of step (b) under conditions wherein the peptide ligase catalyzes the formation of a peptide bond between the amino acids or between the peptide and the amino acid, or between the peptides, thus making at least a dipeptide;
- and
- (d) contacting the peptide of step (c) with the deacetylase of step (b) under conditions wherein the deacetylase catalyzes the deacylation of the peptide, thereby synthesizing a peptide or a polypeptide.
An enzymatic process for synthesizing a peptide comprising the following steps
- (a) providing reaction chamber comprising a peptide ligase and a deacetylase, wherein the peptide ligase and the deacetylase are active under different reaction conditions and the peptide ligase is active after exposure to reaction conditions where the deacetylase is active and the deacetylase is active after exposure to reaction conditions where the peptide ligase is active;
- (b) adding at least two amino acids, at least an amino acid and a peptide, or at least two peptides to the reaction chamber under conditions wherein the peptide ligase is active and the peptide ligase catalyzes the formation of a peptide bond between the amino acids, or between the peptide and the amino acid, or between the peptides;
- and
- (c) changing the conditions in the reaction chamber to conditions wherein the deacetylase is active and the deacetylase catalyzes the deacylation of the peptide formed in step (b).
An enzymatic process for synthesizing a peptide comprising the following steps
- (a) providing reaction chamber comprising a peptide ligase and a deacetylase, wherein the peptide ligase and the deacetylase are active at different temperatures and the peptide ligase is active after exposure to the deacetylase's activity temperature and the deacetylase is active after exposure to the peptide ligase's activity temperature, and at least two amino acids, or at least an amino acid and a peptide, or at least two peptides,
- (b) reacting the reaction chamber under conditions wherein the peptide ligase is active and the deacetylase is inactive and the peptide ligase catalyzes the formation of a peptide bond between the amino acids, or between the peptide and the amino acid, or between the peptides;
- and
- (c) changing the conditions in the reaction chamber to conditions wherein the deacetylase is active and the peptide ligase is inactive and the deacetylase catalyzes the deacylation of the peptide formed in step (b).
A product of manufacture comprising a reaction chamber for synthesizing a peptide comprising a peptide ligase and a deacetylase, wherein the peptide ligase and the deacetylase are active under different reaction conditions and the peptide ligase is active after exposure to reaction conditions where the deacetylase is active and the deacetylase is active after exposure to reaction conditions where the peptide ligase is active.
A product of manufacture for high throughput robotic assays comprising a reaction chamber for synthesizing a peptide comprising an immobilized peptide ligase and an immobilized deacetylase, wherein the peptide ligase and the deacetylase are active under different reaction conditions and the peptide ligase is active after exposure to reaction conditions where the deacetylase is active and the deacetylase is active after exposure to reaction conditions where the peptide ligase is active.
Dependencies are inferred from the claim text.