Abstract: A method for producing a hydroxy-amino ester, or derivative thereof, is provided. A substituted ?-ketodiester having a ketone group and two ester functional groups is contacted with a ketoreductase under conditions permitting the reduction of the ketone group to an alcohol. Only one of the ester functional groups is regioselectively hydrolyzed to the corresponding carboxylic acid, whereby a non-hydrolyzed ester functional group remains. The carboxylic acid is converted to an amine or a derivative thereof to produce a hydroxy-amino ester or derivative thereof. A number of novel hydroxy-amino esters are prepared by the method.

Abstract: Methods for chemically transforming compounds using a mutated enzyme are provided, and more particularly a method for the production of an amino acid from a target 2-ketoacid, the production of an amine from a target ketone and the production of an alcohol from a target ketone. The methods comprise creating a mutated enzyme that catalyzes the reductive amination or transamination of the target 2-ketoacid or ketone or the reduction of the ketone and providing the mutated enzyme in a reaction mixture comprising the target 2-ketoacid or ketone under conditions sufficient to permit the formation of the desired amino acid, amine or alcohol to thereby produce the amino acid, amine or alcohol.

Abstract: Hydroxy-amino acids are provided and are prepared by contacting a substituted ?-ketodiester having a ketone group and two ester functional groups with a ketoreductase under conditions permitting the reduction of the keytone group to an alcohol. Only one of the ester functional groups is regioselectively hydrolyzed to the corresponding carboxylic acid, whereby a non-hydrolyzed ester functional group remains. The carboxylic acid is converted to an amine to produce a hydroxy-amino acid.

Abstract: A methods for producing a hydroxy-amino acid or derivative thereof, such as statine, phenylstatine or isostatine, is provided. A substituted &bgr;-ketodiester having a ketone group and two ester functional groups is contacted with a ketoreductase under conditions permitting the reduction of the ketone group to an alcohol. Only one of the ester functional groups is regioselectively hydrolyzed to the corresponding carboxylic acid, whereby a non-hydrolyzed ester functional group remains. Either the carboxylic acid or the non-hydrolyzed ester functional group is converted to an amine or a derivative thereof to produce a hydroxy-amino acid or derivative thereof.

Abstract: Methods for producing single diastereomers of isoleucine in high stereochemical purity are provided. D-isoleucine is produced by converting (R)-2-methylbutyraldehyde to a diastereomeric mixture of D-isoleucine hydantoin and L-allo-isoleucine hydantoin (5S-[(R)-1-methylpropyl]hydantoin) under conditions whereby no significant racemization of the chiral center in (R)-2-methylbutyraldehyde occurs, followed by contacting said diastereomeric hydantoin mixture with a D-hydantoinase to stereoselectively hydrolyze any D-isoleucine hydantoin in the mixture to the corresponding N-carbamoyl-D-isoleucine, preferably under conditions permitting the simultaneous epimerization of the chiral center at C-5 of the hydantoin. The simultaneous epimerization permits the reaction to be carried out to substantial completion so that the diastereomeric hydantoin mixture is converted to N-carbamoyl-D-isoleucine in high yield. The N-carbamoyl-D-isoleucine is then decarbamoylated to produce D-isoleucine.

Abstract: A method for producing D-allo-isoleucine is provided. The method comprises converting L-isoleucine to the corresponding hydantoin. A mixture containing the hydantoin is contacted with a D-hydantoinase to stereoselectively hydrolyze any D-allo-isoleucine hydantoin in the mixture to the corresponding N-carbamoyl-D-allo-isoleucine. The N-carbamoyl-D-allo-isoleucine is decarbamoylated to produce D-allo-isoleucine. Preferably the contacting of the hydantoin with a D-hydantoinase is carried out under conditions permitting the simultaneous epimerization of the chiral center at C-5 of the hydantoin.

Abstract: A synthetic nucleic acid sequence is disclosed, comprising a non-naturally occurring polymer of nucleic acids, having a biological function encoded by the sequence and known from a starting nucleic acid sequence, and having a difference in sequence of at least about 5% between the nucleic acids of the synthetic sequence and the starting sequence. The difference between the nucleic acid sequences results in a different free energy of folding for the synthetic sequence as compared to the starting sequence, such that the synthetic sequence would be expressed better in a selected heterologous host cell than the starting sequence would be if expressed in the same heterologous host cell.

Abstract: An enzyme that cleaves 1,3-dicarbonyl compounds by incorporating molecular oxygen into the substrate in amounts equimolar to the amount of substrate cleaved. The enzyme from Acinetobacter johnsonii is a multimer of about 67 kilodaltons made up of subunits of about 16.6 kilodaltons. Nucleotide and amino acid sequence analysis of a cloned A. johnsonii subunit indicates little homology with other proteins. Native or recombinant enzyme can be used to decontaminate or detoxify acetylacetone or related diketones.

Abstract: A method of making a synthetic nucleic acid sequence comprises providing a starting nucleic acid sequence, which optionally encodes an amino acid sequence, and determining the predicted &Dgr;Gfolding of the sequence. The starting nucleic acid sequence can be a naturally occurring sequence or a non-naturally occurring sequence. The starting nucleic acid sequence is modified by replacing at least one codon from the starting nucleic acid sequence with a different corresponding codon to provide a modified nucleic acid sequence. As used herein, a “different corresponding codon” refers to a codon which does not have the identical nucleotide sequence, but which encodes the identical amino acid. The predicted &Dgr;Gfolding of the modified nucleic acid sequence is determined and compared with the &Dgr;Gfolding of the starting nucleic acid sequence.

Abstract: The disclosure describes novel precursors for the preparation of chiral 1,3-aminoalcohols. The precursors are chiral 4-hydroxycarboxamides, 4-hydroxyhydroxamic acids, or 4-hydroxyhydrazides produced from chiral gamma-lactones, which in turn are derived from 1,4-diols by stereoselective oxidation. The chiral 4-hydroxycarboxamides, 4-hydroxyhydroxamic acids, or 4-hydroxyhydrazides are converted into chiral 1,3-aminoalcohols by stereospecific rearrangement.

Abstract: The disclosure describes new compositions of matter useful for the preparation of optically-active vicinal aminoalcohols. The compositions are chiral .beta.-hydroxycarboxamides, .beta.-hydroxyhydraxides, and .beta.-hydroxyhydroxamic acids.

Abstract: The disclosure describes a method for the preparation of chiral 1,3-aminoalcohols in high optical purity. The method combines the stereoselective oxidation of a 1,4-diol to a gamma-lactone using an alcohol dehydrogenase, the conversion of the gamma-lactone to the corresponding 4-hydroxyamide, 4-hydroxyhydroxamic acid, or 4-hydroxyhydrazide, and stereospecific rearrangement of the 4-hydroxyamide, 4-hydroxyhydroxamic acid, or 4-hydroxyhydrazide to the corresponding chiral 1,3-aminoalcohol.

Abstract: A method for quantitating L-homocysteine and/or L-methionine in a solution involves contacting a solution containing L-homocysteine and/or L-methionine with a reagent comprising methionine gamma-lyase and a cofactor capable of forming a Schiff base with the L-methionine and/or L-homocysteine for a time sufficient to catalyze the conversion of L-homocysteine and/or L-methionine to 2-ketobutyrate. The amount of 2-ketobutyrate formed is determined, and the amount of L-homocysteine and/or L-methionine present in the original solution can be determined based on the amount of 2-ketobutyrate formed. A composition for measuring the amount of L-homocysteine and/or L-methionine in a solution comprises methionine gamma-lyase, a cofactor capable of forming a Schiff base with the L-methionine and/or L-homocysteine and at least one 2-ketobutyrate detecting agent, but is substantially free of L-methionine, L-homocysteine, 2-ketobutyrate, pyruvate and mercury.

Abstract: The disclosure describes a method for the preparation of chiral vicinal aminoalcohols in high optical purity. The method combines the stereoselective reduction of the keto group of a .beta.-ketoacid, .beta.-keotester, or derivative with the stereospecific rearrangement of the corresponding amide, hyroxamic acid, or hydrazide to produce chiral vicinal aminoalcohols with control of stereochemistry at both chiral centers.