Abstract: The present disclosure provides methods of producing commodity products, the methods involving culturing a host cell that is genetically modified to produce a uronate dehydrogenase (UDH) that converts a sugar acid to its corresponding 1,5-aldonolactone, that uses NADP+ or NAD+ as a cofactor, and that produces NADPH or NADH, respectively, where the host cell coexpresses an endogenous or a heterologous reductase that utilizes the produced NADPH or NADH to generate the commodity product or a precursor thereof. The present disclosure provides a method of producing downstream products of glycerol and pyruvate in a genetically modified microbial host cell, the method involving culturing a genetically modified microbial host cell of the present disclosure in a culture medium comprising D-galacturonic acid. The present disclosure provides variant UDH polypeptides that utilize NADP+, nucleic acids encoding the variant UDH polypeptides; and host cells genetically modified with the nucleic acids.

Abstract: The present disclosure provides a variant tyrosine hydroxylase that provides for increased production of L-DOPA in a host cell that expresses the tyrosine hydroxylase. The present disclosure provides nucleic acids encoding the variant tyrosine hydroxylase, and host cells genetically modified with the nucleic acids. The present disclosure provides methods of making L-DOPA in a host cell. The present disclosure provides methods of making a benzylisoquinoline alkaloid (BIA), or a BIA precursor. The present disclosure provides methods of detecting L-DOPA level in a cell. The present disclosure provides methods of identifying tyrosine hydroxylase variants that provide for increased L-DOPA production; and methods of identifying gene products that provide for increased tyrosine production.

Abstract: The present disclosure provides methods of producing commodity products, the methods involving culturing a host cell that is genetically modified to produce a uronate dehydrogenase (UDH) that converts a sugar acid to its corresponding 1,5-aldonolactone, that uses NADP+ or NAD+ as a cofactor, and that produces NADPH or NADH, respectively, where the host cell coexpresses an endogenous or a heterologous reductase that utilizes the produced NADPH or NADH to generate the commodity product or a precursor thereof. The present disclosure provides a method of producing downstream products of glycerol and pyruvate in a genetically modified microbial host cell, the method involving culturing a genetically modified microbial host cell of the present disclosure in a culture medium comprising D-galacturonic acid. The present disclosure provides variant UDH polypeptides that utilize NADP+, nucleic acids encoding the variant UDH polypeptides; and host cells genetically modified with the nucleic acids.

Abstract: The present invention provides methods of producing a product or product precursor of a biosynthetic pathway in a genetically modified host cell. The present invention also provides genetically modified host cells comprising nucleic acids encoding a scaffold polypeptide and nucleic acids comprising nucleotide sequences encoding two or more enzymes in a biosynthetic pathway. The present invention further provides nucleic acids comprising nucleotide sequences encoding scaffold polypeptides, for use in a subject method.

Abstract: The present disclosure provides a variant tyrosine hydroxylase that provides for increased production of L-DOPA in a host cell that expresses the tyrosine hydroxylase. The present disclosure provides nucleic acids encoding the variant tyrosine hydroxylase, and host cells genetically modified with the nucleic acids. The present disclosure provides methods of making L-DOPA in a host cell. The present disclosure provides methods of making a benzylisoquinoline alkaloid (BIA), or a BIA precursor. The present disclosure provides methods of detecting L-DOPA level in a cell. The present disclosure provides methods of identifying tyrosine hydroxylase variants that provide for increased L-DOPA production; and methods of identifying gene products that provide for increased tyrosine production.

Abstract: The present invention provides for a recombinant nucleic acid comprising a nucleotide sequence comprising a plurality of constructs, wherein each construct independently comprises a nucleotide sequence of interest flanked by a pair of recombinase recognition sequences. Each pair of recombinase recognition sequences is recognized by a distinct recombinase. Optionally, each construct can, independently, further comprise one or more genes encoding a recombinase capable of recognizing the pair of recombinase recognition sequences of the construct. The recombinase can be an orthogonal (non-cross reacting), site-selective recombinase (OSSR).

Abstract: The present invention provides methods of producing a product or product precursor of a biosynthetic pathway in a genetically modified host cell. The present invention also provides genetically modified host cells comprising nucleic acids encoding a scaffold polypeptide and nucleic acids comprising nucleotide sequences encoding two or more enzymes in a biosynthetic pathway. The present invention further provides nucleic acids comprising nucleotide sequences encoding scaffold polypeptides, for use in a subject method.

Abstract: The present invention provides methods of producing a product or product precursor of a biosynthetic pathway in a genetically modified host cell. The present invention also provides genetically modified host cells comprising nucleic acids encoding a scaffold polypeptide and nucleic acids comprising nucleotide sequences encoding two or more enzymes in a biosynthetic pathway. The present invention further provides nucleic acids comprising nucleotide sequences encoding scaffold polypeptides, for use in a subject method.

Abstract: The present invention provides for a recombinant nucleic acid comprising a nucleotide sequence comprising a plurality of constructs, wherein each construct independently comprises a nucleotide sequence of interest flanked by a pair of recombinase recognition sequences. Each pair of recombinase recognition sequences is recognized by a distinct recombinase. Optionally, each construct can, independently, further comprise one or more genes encoding a recombinase capable of recognizing the pair of recombinase recognition sequences of the construct. The recombinase can be an orthogonal (non-cross reacting), site-selective recombinase (OSSR).

Abstract: The present invention provides methods of producing a product or product precursor of a biosynthetic pathway in a genetically modified host cell. The present invention also provides genetically modified host cells comprising nucleic acids encoding a scaffold polypeptide and nucleic acids comprising nucleotide sequences encoding two or more enzymes in a biosynthetic pathway. The present invention further provides nucleic acids comprising nucleotide sequences encoding scaffold polypeptides, for use in a subject method.