Certain embodiments provide a method for preparing a biochemical product (e.g., phenol, catechol, or muconic acid, or a salt thereof). For example, such methods include contacting a recombinant host having two or more recombinant pathways with a fermentable carbon source and growing the recombinant cell for a time sufficient to synthesize the product. In certain embodiments, each recombinant pathway: 1) is capable of producing the same final biochemical product; 2) comprises at least one gene encoding a polypeptide; 3) is derived from a different endogenous metabolite as its immediate precursor; and 4) converges to the same final product or the same intermediate metabolite.

Biosensors including a nucleic acid encoding a PcaU protein, a PobR protein, a CatM protein, a PcaR protein, or a TphR protein are provided. In some examples, the biosensors include a promoter regulated by the sensed molecule operably linked to a reporter gene. The biosensors may be included in a vector or in cells including one or more of the biosensors or vectors. Modified chorismate pyruvate lyase (UbiC) and modified paraoxonase (PON1) proteins including one or more amino acid substitutions are provided. Finally, methods of selecting biocatalysts with increased activity including transforming a library of cells expressing a biosensor with one or more nucleic acids encoding one or more mutations in a gene involved in a biosynthesis pathway, determining activity of the reporter protein; and selecting a cell with increased reporter protein activity as expressing a biocatalyst with increased activity are provided.

A method of producing para-hydroxybenzoic acid (pHBA) or a derivative thereof includes culturing the recombinant microorganism in a fermentation broth, wherein said recombinant microorganism comprising a genetically engineered pathway expressing at least one nucleic acid sequence encoding a polypeptide selected from: an exogenous chorismate pyruvate lyase of EC 5.4.4.2 or EC 4.1.3.40; an exogenous 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase of EC 4.1.2.15, or EC 2.5.1.54; an exogenous shikimate kinase of EC 2.7.1.71; or an exogenous 3-dehydroquinate dehydratase (DHQ) of EC 4.2.1.10; adding a carbon source to the fermentation broth; and isolating the pHBA from the fermentation broth.

A method for conferring tolerance to a 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor herbicide in a plant by expressing one or more polypeptide components of an exogenous plastoquinone-9 pathway in the plant. Nucleic acids, vectors, transgenic cells and transgenic plants useful in such a method are also disclosed.

Provided is a microorganism that is able to efficiently produce protocatechuic acid or a salt thereof by using a saccharide as a raw material, and a method of efficiently producing protocatechuic acid or a salt thereof by using the microorganism. Provided is a transformant having protocatechuic acid producing ability, subjected to modifications of enhancement of 3-dehydroshikimate dehydratase activity; enhancement of chorismate pyruvate lyase activity; and enhancement of 4-hydroxybenzoate hydroxylase activity. Also provided is a method of producing protocatechuic acid or a salt thereof, including the step of culturing the transformant in a reaction solution containing a saccharide so as to cause the transformant to produce protocatechuic acid or a salt thereof.

The mutant chorismate-pyruvate lyase (A) or (B) as described below is capable of producing 4-hydroxybenzoic acid or a salt thereof with sufficient practical efficiency. (A) A mutant chorismate-pyruvate lyase obtained by replacing the valine at position 80 in a chorismate-pyruvate lyase (ubiC) from Pantoea ananatis consisting of the amino acid sequence of SEQ ID NO: 1 with one or more other amino acids. (B) A mutant chorismate-pyruvate lyase obtained by replacing an amino acid in another chorismate-pyruvate lyase, the amino acid being at a position enzymologically homologous with that of the above valine, with one or mere other amino acids.

Certain embodiments provide a method for preparing a biochemical product (e.g., phenol, catechol, or muconic acid, or a salt thereof). For example, such methods include contacting a recombinant host having two or more recombinant pathways with a fermentable carbon source and growing the recombinant cell for a time sufficient to synthesize the product. In certain embodiments, each recombinant pathway: 1) is capable of producing the same final biochemical product; 2) comprises at least one gene encoding a polypeptide; 3) is derived from a different endogenous metabolite as its immediate precursor; and 4) converges to the same final product or the same intermediate metabolite.

The invention provides non-naturally occurring microbial organisms having a (2-hydroxy-3-methyl-4-oxobutoxy)phosphonate pathway, p-toluate pathway, and/or terephthalate pathway. The invention additionally provides methods of using such organisms to produce (2-hydroxy-3-methyl-4-oxobutoxy)phosphonate pathway, p-toluate pathway or terephthalate pathway.

The mutant chorismate-pyruvate lyase (A) or (B) as described below is capable of producing 4-hydroxybenzoic acid or a salt thereof with sufficient practical efficiency.

(A) A mutant chorismate-pyruvate lyase obtained by replacing the valine at position 80 in a chorismate-pyruvate lyase (ubiC) from Pantoea ananatis consisting of the amino acid sequence of SEQ ID NO: 1 with one or more other amino acids.

(B) A mutant chorismate-pyruvate lyase obtained by replacing an amino acid in another chorismate-pyruvate lyase, the amino acid being at a position enzymologically homologous with that of the above valine, with one or mere other amino acids.

Provided is a microorganism that is able to efficiently produce protocatechuic acid or a salt thereof by using a saccharide as a raw material, and a method of efficiently producing protocatechuic acid or a salt thereof by using the microorganism.

Provided is a transformant having protocatechuic acid producing ability, subjected to modifications (A), (B), and (C) below: (A) enhancement of 3-dehydroshikimate dehydratase activity; (B) enhancement of chorismate pyruvate lyase activity; and (C) enhancement of 4-hydroxybenzoate hydroxylase activity.

Also provided is a method of producing protocatechuic acid or a salt thereof, including the step of culturing the transformant in a reaction solution containing a saccharide so as to cause the transformant to produce protocatechuic acid or a salt thereof.