This invention relates to the bioproduction of substituted or unsubstituted phenylacetaldehyde, 2-phenylethanol, phenylacetic acid or phenylethylamine by subjecting a starting material comprising glucose, L-phenylalanine, substituted L-phenylalanine, styrene or substituted styrene to a plurality of enzyme catalyzed chemical transformations in a one-pot reaction system, using recombinant microbial cells overexpressing the enzymes. To produce phenylacetaldehyde from styrene, the cells are modified to overexpress styrene monooxygenase (SMO) and styrene oxide isomerase (SOI). To produce phenylacetic acid from styrene, SMO, SOI and aldehyde dehydrogenase are overexpressed. Alternatively, to produce 2-phenylethanol, SMO, SOI and aldehyde reductase or alcohol dehydrogenase are overexpressed, while to produce phenylethylamine, SMO, SOI and transaminase are overexpressed.

This invention relates to the bioproduction of substituted or unsubstituted phenylacetaldehyde, 2-phenylethanol, phenylacetic acid or phenylethylamine by subjecting a starting material comprising glucose, L-phenylalanine, substituted L-phenylalanine, styrene or substituted styrene to a plurality of enzyme catalyzed chemical transformations in a one-pot reaction system, using recombinant microbial cells overexpressing the enzymes. To produce phenylacetaldehyde from styrene, the cells are modified to overexpress styrene monooxygenase (SMO) and styrene oxide isomerase (SOI). To produce phenylacetic acid from styrene, SMO, SOI and aldehyde dehydrogenase are overexpressed. Alternatively, to produce 2-phenylethanol, SMO, SOI and aldehyde reductase or alcohol dehydrogenase are overexpressed, while to produce phenylethylamine, SMO, SOI and transaminase are overexpressed.

The technical field of preparation of organic compounds, and particularly to yeasts producing tyrosol or hydroxytyrosol and construction methods thereof. PcAAS and ADH-encoding DNA sequences are introduced into the yeast strain BY4741, to obtain a PcAAS-ADH recombinant yeast producing tyrosol. A PDC1 knockout cassette and a TyrA expression cassette are introduced into the PcAAS-ADH recombinant yeast to obtain a PcAAS-ADH-ΔPDC1-TyrA recombinant yeast producing tyrosol. A HpaBC encoding DNA sequence is introduced into the PcAAS-ADH-ΔPDC1-TyrA recombinant yeast, to obtain a PcAAS-ADH-HpaBC-ΔPDC1-TyrA recombinant yeast producing hydroxytyrosol. The construction of a tyrosol or hydroxytyrosol biosynthesis pathway in the yeast strain BY4741 enhances the production of tyrosol or hydroxytyrosol.

The technical field of preparation of organic compounds, and particularly to yeasts producing tyrosol or hydroxytyrosol and construction methods thereof. PcAAS and ADH-encoding DNA sequences are introduced into the yeast strain BY4741, to obtain a PcAAS-ADH recombinant yeast producing tyrosol. A PDC1 knockout cassette and a TyrA expression cassette are introduced into the PcAAS-ADH recombinant yeast to obtain a PcAAS-ADH-ΔPDC1-TyrA recombinant yeast producing tyrosol. A HpaBC encoding DNA sequence is introduced into the PcAAS-ADH-ΔPDC1-TyrA recombinant yeast, to obtain a PcAAS-ADH-HpaBC-ΔPDC1-TyrA recombinant yeast producing hydroxytyrosol. The construction of a tyrosol or hydroxytyrosol biosynthesis pathway in the yeast strain BY4741 enhances the production of tyrosol or hydroxytyrosol.

Polyketide synthases are engineered to produce lactones. In the first module, an acyltransferase is swapped and in the second module a reductive loop is swapped. With another acyltransferase swap in the second module, we can programmably produce the non-methylated delta lactone.

A process for preparing phenylacetic acid is provided. The process essentially includes the steps of providing one or more yeast strains belonging to the genus Yarrowia and mutants thereof, providing a culture medium including phenylalanine, transforming phenylalanine into phenylacetic acid by fermentation of the one or more yeast strains in the culture medium, the phenylacetic acid being contained in a fermentation broth obtained by the fermentation, and isolating the phenylacetic acid from the fermentation broth.

Described herein are engineered metabolic pathways in recombinant microorganism host cells which result in the production of 2-phenylethanol or 2-phenylacetic acid. Also described herein are methods of using the recombinant microorganisms for the production of 2-phenylethanol or 2-phenylacetic acid.

Described herein are engineered metabolic pathways in recombinant microorganism host cells which result in the production of 2-phenylethanol or 2-phenylacetic acid. Also described herein are methods of using the recombinant microorganisms for the production of 2-phenylethanol or 2-phenylacetic acid.

Described herein are prenyltransferases including non-natural variants thereof having at least one amino acid substitution as compared to its corresponding natural or unmodified prenyltransferases and that are capable of at least two-fold greater rate of formation of cannabinoids such as cannabigerolic acid, cannabigerovarinic acid, cannabigerorcinic acid, and cannabigerol, as compared to a wild type control. Prenyltransferase variants also demonstrated regioselectivity to desired cannabinoid isomers such as CDBA (3-GOLA), 3-GDVA, 3-GOSA, and CBG (2-GOL). The prenyltransferase variants can be used to form prenylated aromatic compounds, and can be expressed in an engineered microbe having a pathway to such compounds, which include 3-GOLA, 3-GDVA, 3-GOSA, and CBG. 3-GOLA can be used for the preparation of cannabigerol (CBG), which can be used in therapeutic compositions.

Described herein are prenyltransferases including non-natural variants thereof having at least one amino acid substitution as compared to its corresponding natural or unmodified prenyltransferases and that are capable of at least two-fold greater rate of formation of cannabinoids such as cannabigerolic acid, cannabigerovarinic acid, cannabigerorcinic acid, and cannabigerol, as compared to a wild type control. Prenyltransferase variants also demonstrated regioselectivity to desired cannabinoid isomers such as CDBA (3-GOLA), 3-GDVA, 3-GOSA, and CBG (2-GOL). The prenyltransferase variants can be used to form prenylated aromatic compounds, and can be expressed in an engineered microbe having a pathway to such compounds, which include 3-GOLA, 3-GDVA, 3-GOSA, and CBG. 3-GOLA can be used for the preparation of cannabigerol (CBG), which can be used in therapeutic compositions.