Abstract: The present disclosure provides novel engineered target-biomolecule-producing bacterial strains and methods of producing the same. To engineer bacterial strains capable of producing substantial levels of a target biomolecule, the methods may implement the use of metabolic modeling and machine learning methods. The methods and bacterial strains produced by the methods may be implemented in further optimizing a biosynthetic pathway, e.g., to improve the production of a target biomolecule of interest, e.g., an amino acid, such as threonine.

Abstract: Described herein are small peptide domains and consensus sequences that bind small target molecules of industrial importance, e.g., metals such as nickel, ? carotene, and isoflavones such as genistein. Also described are fusion proteins containing such binding domains fused to proteins or to peptide domains like GST or CBD that bind other ligands and can be used to immobilize the target binding domain on a support. One class of fusion proteins that is useful in industrial settings are fusions that contain concatemers of target binding domains, which increases the binding equivalents per molecule.

Abstract: Described herein are small peptide domains and consensus sequences that bind small target molecules of industrial importance, e.g., metals such as nickel, ? carotene, and isoflavones such as genistein. Also described are fusion proteins containing such binding domains fused to proteins or to peptide domains like GST or CBD that bind other ligands and can be used to immobilize the target binding domain on a support. One class of fusion proteins that is useful in industrial settings are fusions that contain concatemers of target binding domains, which increases the binding equivalents per molecule.

Abstract: Described herein are small peptide domains and consensus sequences that bind small target molecules of industrial importance, e.g., metals such as nickel, ? carotene, and isoflavones such as genistein. Also described are fusion proteins containing such binding domains fused to proteins or to peptide domains like GST or CBD that bind other ligands and can be used to immobilize the target binding domain on a support. One class of fusion proteins that is useful in industrial settings are fusions that contain concatemers of target binding domains, which increases the binding equivalents per molecule.

Abstract: Described herein are small peptide domains and consensus sequences that bind small target molecules of industrial importance, e.g., metals such as nickel, ? carotene, and isoflavones such as genistein. Also described are fusion proteins containing such binding domains fused to proteins or to peptide domains like GST or CBD that bind other ligands and can be used to immobilize the target binding domain on a support. One class of fusion proteins that is useful in industrial settings are fusions that contain concatemers of target binding domains, which increases the binding equivalents per molecule.

Abstract: Described herein are small peptide domains and consensus sequences that bind small target molecules of industrial importance, e.g., metals such as nickel, ? carotene, and isoflavones such as genistein. Also described are fusion proteins containing such binding domains fused to proteins or to peptide domains like GST or CBD that bind other ligands and can be used to immobilize the target binding domain on a support. One class of fusion proteins that is useful in industrial settings are fusions that contain concatemers of target binding domains, which increases the binding equivalents per molecule.

Abstract: A method of converting hydroxymethylfurfural and is derivative species into hydroxymethylfurfural oxidation products is disclosed. The method includes contacting the hydroxymethylfurfural species in a mixture with an enzyme that oxidizes the hydroxymethylfurfural species while controlling hydrogen peroxide in the mixture. In one exemplary embodiment the enzyme is chloroperoxidase and the hydrogen peroxide is metered into the mixture to predominantly and selectively make at least one of formylfuran carboxylic acid or furan dicarboxylic acid. In another embodiment the enzyme is aryl alcohol oxidase and catalase is included in the mixture to remove unwanted hydrogen peroxide by product and the reaction predominantly makes at least one of dimethylfuran or formylfuran carboxylic acid. When the predominant product is a carboxylic acid or furan dicarboxylic acid, it can be recovered in substantially pure form by acid precipitation.

Abstract: The invention provides methods to increase the production of an amino acid from Corynebacterium species by way of the amplification of amino acid biosynthetic pathway genes in a host cell chromosome. The invention also provides novel processes for the production of an amino acid by way of the amplification of amino acid biosynthetic pathway genes in a host cell chromosome and/or by increasing promoter strength. In a preferred embodiment, the invention provides processes to increase the production of L-lysine in Corynebacterium glutamicum by way of the amplification of L-lysine biosynthetic pathway genes in a host cell chromosome. The invention also provides novel isolated nucleic acid molecules for L-lysine biosynthetic pathway genes of Corynebacterium glutamicum.

Abstract: Methods and compositions for increased production of amino acids from C. glutamicum using sucrose as a carbon source are described. In one aspect, increased production of L-lysine from C. glutamicum is accomplished by using a strain having a mutation in the ptsF gene encoding fructose-PTS enzyme that attenuates or blocks fructose import into the cell when such strain is grown on media containing sucrose as a carbon source and production is increased by providing glucose isomerase in the fermentation media. The glucose isomerase may be exogenously added or expressed in the strain and exported into the media. In certain embodiments the media also contain an invertase. In another aspect increased production of L-lysine is accomplished by making a C. glutamicum strain having the ptsF mutation and a second mutation in a fructose exporter function. The dual mutation retains imported fructose in the cell.

Abstract: The invention provides methods to increase the production of an amino acid from Corynebacterium species by way of the amplification of amino acid biosynthetic pathway genes in a host cell chromosome. The invention also provides novel processes for the production of an amino acid by way of the amplification of amino acid biosynthetic pathway genes in a host cell chromosome and/or by increasing promoter strength. In a preferred embodiment, the invention provides processes to increase the production of L-lysine in Corynebacterium glutamicum by way of the amplification of L-lysine biosynthetic pathway genes in a host cell chromosome. The invention also provides novel isolated nucleic acid molecules for L-lysine biosynthetic pathway genes of Corynebacterium glutamicum.

Abstract: The invention provides methods to increase the production of an amino acid from Corynebacterium species by way of the amplification of amino acid biosynthetic pathway genes in a host cell chromosome. Amplification may be by integration of one or more copies of a gene or genes into a host cell chromosome. One gene that may be incorporated is the gene ORF2, which encodes an unnamed hypothetical protein and which may be obtained from Corynebacterium glutamicum. The invention also provides novel isolated nucleic acid molecules for L-lysine biosynthetic pathway genes of Corynebacterium glutamicum.

Abstract: Methods and compositions for increased production of amino acids from C. glutamicum using sucrose as a carbon source are described. In one aspect, increased production of L-lysine from C. glutamicum is accomplished by using a strain having a mutation in the ptsF gene encoding fructose-PTS enzyme that attenuates or blocks fructose import into the cell when such strain is grown on media containing sucrose as a carbon source and production is increased by providing glucose isomerase in the fermentation media. The glucose isomerase may be exogenously added or expressed in the strain and exported into the media. In certain embodiments the media also contain an invertase. In another aspect increased production of L-lysine is accomplished by making a C. glutamicum strain having the ptsF mutation and a second mutation in a fructose exporter function. The dual mutation retains imported fructose in the cell.

Abstract: A method of converting hydroxymethylfurfural and is derivative species into hydroxymethylfurfural oxidation products is disclosed. The method includes contacting the hydroxymethylfurfural species in a mixture with an enzyme that oxidizes the hydroxymethylfurfural species while controlling hydrogen peroxide in the mixture. In one exemplary embodiment the enzyme is chloroperoxidase and the hydrogen peroxide is metered into the mixture to predominantly and selectively make at least one of formylfuran carboxylic acid or furan dicarboxylic acid. In another embodiment the enzyme is aryl alcohol oxidase and catalase is included in the mixture to remove unwanted hydrogen peroxide by product and the reaction predominantly makes at least one of dimethylfuran or formylfuran carboxylic acid. When the predominant product is a carboxylic acid or furan dicarboxylic acid, it can be recovered in substantially pure form by acid precipitation.

Abstract: The present invention relates to a mutated pyruvate carboxylase gene from Corynebacterium. The mutant pyruvate carboxylase gene encodes a pyruvate carboxylase enzyme which is resistant to feedback inhibition from aspartic acid. The present invention also relates to a method of replacing the wild-type pyruvate carboxylase gene in Corynebacterium with this feedback-resistant pyruvate carboxylase gene. The present invention further relates to methods of the production of amino acids, preferably lysine, comprising the use of this mutant pyruvate carboxylase enzyme in microorganisms.

Abstract: Methods for cell-free production of glucosamine from starch, maltodextrin or glycogen or from fructose and a source of amino groups are disclosed. Also disclosed are cellular extracts comprising glucosamine-6-phosphate synthase activity, as well as a cellular extract comprising glucosamine-6-phosphate deaminase.

Abstract: The present invention relates to a mutated pyruvate carboxylase gene from Corynebacterium. The mutant pyruvate carboxylase gene encodes a pyruvate carboxylase enzyme which is resistant to feedback inhibition from aspartic acid. The present invention also relates to a method of replacing the wild-type pyruvate carboxylase gene in Corynebacterium with this feedback-resistant pyruvate carboxylase gene. The present invention further relates to methods of the production of amino acids, preferably lysine, comprising the use of this mutant pyruvate carboxylase enzyme in microorganisms.

Abstract: The invention provides methods to increase the production of an amino acid from Corynebacterium species by way of the amplification of amino acid biosynthetic pathway genes in a host cell chromosome. In a preferred embodiment, the invention provides methods to increase the production of L-lysine in Corynebacterium glutamicum by way of the amplification of L-lysine biosynthetic pathway genes in a host cell chromosome. The invention also provides novel processes for the production of an amino acid by way of the amplification of amino acid biosynthetic pathway genes in a host cell chromosome and/or by increasing promoter strength. In a preferred embodiment, the invention provides processes to increase the production of L-lysine in Corynebacterium glutamicum by way of the amplification of L-lysine biosynthetic pathway genes in a host cell chromosome.

Abstract: The present invention relates, in general, to a method of producing L-amino acids comprising culturing altered bacterial cells having increased amounts of NADPH as compared to unaltered bacterial cells whereby L-amino acids yields from said altered bacterial cells are greater than yields from unaltered bacterial cells. The invention also relates to a gene encoding phosphoglucoisomerase.

Abstract: The invention provides methods to increase the production of an amino acid from Corynebacterium species by way of the amplification of amino acid biosynthetic pathway genes in a host cell chromosome. The invention also provides novel processes for the production of an amino acid by way of the amplification of amino acid biosynthetic pathway genes in a host cell chromosome and/or by increasing promoter strength. In a preferred embodiment, the invention provides processes to increase the production of L-lysine in Corynebacterium glutamicum by way of the amplification of L-lysine biosynthetic pathway genes in a host cell chromosome. The invention also provides novel isolated nucleic acid molecules for L-lysine biosynthetic pathway genes of Corynebacterium glutamicum.

Abstract: The present invention relates, in general, to a method of producing L-amino acids comprising culturing altered bacterial cells having increased amounts of NADPH as compared to unaltered bacterial cells whereby L-amino acids yields from said altered bacterial cells are greater than yields from unaltered bacterial cells. The invention also relates to a gene encoding phosphoglucoisomerase.