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.

Provided herein is a non-naturally occurring microbial organism having a methanol metabolic pathway that can enhance the availability of reducing equivalents in the presence of methanol. Such reducing equivalents can be used to increase the product yield of organic compounds produced by the microbial organism, such as 1,2-propanediol, n-propanol, 1,3-propanediol or glycerol. Also provided herein are methods for using such an organism to produce 1,2-propanediol, n-propanol, 1,3-propanediol or glycerol.

Provided herein is a non-naturally occurring microbial organism having a methanol metabolic pathway that can enhance the availability of reducing equivalents in the presence of methanol. Such reducing equivalents can be used to increase the product yield of organic compounds produced by the microbial organism, such as 1,2-propanediol, n-propanol, 1,3-propanediol or glycerol. Also provided herein are methods for using such an organism to produce 1,2-propanediol, n-propanol, 1,3-propanediol or glycerol.

Described herein are non-natural NAD.sup.+-dependent alcohol dehydrogenases (ADHs) capable of at least two fold greater conversion of methanol or ethanol to formaldehyde or acetaldehyde, respectively, as compared to its unmodified counterpart. Nucleic acids encoding the non-natural alcohol dehydrogenases, as well as expression constructs including the nucleic acids, and engineered cells comprising the nucleic acids or expression constructs are described. Also described are engineered cells expressing a non-natural NAD.sup.+-dependent alcohol dehydrogenase, optionally include one or more additional metabolic pathway transgene(s), methanol metabolic pathway genes, target product pathway genes, cell culture compositions including the cells, methods for promoting production of the target product or intermediate thereof from the cells, compositions including the target product or intermediate, and products made from the target product or intermediate.

Described herein are non-natural NAD.sup.+-dependent alcohol dehydrogenases (ADHs) capable of at least two fold greater conversion of methanol or ethanol to formaldehyde or acetaldehyde, respectively, as compared to its unmodified counterpart. Nucleic acids encoding the non-natural alcohol dehydrogenases, as well as expression constructs including the nucleic acids, and engineered cells comprising the nucleic acids or expression constructs are described. Also described are engineered cells expressing a non-natural NAD.sup.+-dependent alcohol dehydrogenase, optionally include one or more additional metabolic pathway transgene(s), methanol metabolic pathway genes, target product pathway genes, cell culture compositions including the cells, methods for promoting production of the target product or intermediate thereof from the cells, compositions including the target product or intermediate, and products made from the target product or intermediate.

A variety of methods and systems for screening biocatalysts are disclosed, including, in one embodiment, a screening method for identifying engineered biocatalysts, including reacting an olefin with water in the presence of an engineered biocatalyst to produce at least a fatty alcohol having from 4 carbons to 24 carbons; reacting at least a portion of the fatty alcohol with oxygen in the present of a fatty alcohol oxidase to produce a fatty aldehyde and hydrogen peroxide, the fatty aldehyde having from 4 carbons to 24 carbons; and measuring activity of the engineered biocatalyst.

A variety of methods and systems for screening biocatalysts are disclosed, including, in one embodiment, a screening method for identifying engineered biocatalysts, including reacting an olefin with water in the presence of an engineered biocatalyst to produce at least a fatty alcohol having from 4 carbons to 24 carbons; reacting at least a portion of the fatty alcohol with oxygen in the present of a fatty alcohol oxidase to produce a fatty aldehyde and hydrogen peroxide, the fatty aldehyde having from 4 carbons to 24 carbons; and measuring activity of the engineered biocatalyst.

A system for the production of high value chemicals includes (a) an input selected from the group consisting of ethylene glycol, glycerol, ethanol methanol or a combination thereof. In addition, the system includes (b) an oxidation biocatalyst including an alcohol oxidase, a copper radical oxidase, a glycerol oxidase, an alditol oxidase or a combination thereof. Further, the system includes (c) an oxidized intermediate. The system also includes (d) a finishing catalyst including a supported metal catalyst, a carboligating catalyst, an amine oxidase, a glyoxalase, an acid catalyst, a base catalyst, an isomerization catalyst or a combination thereof. Still further, the system includes (e) an output.

A system for the production of high value chemicals includes (a) an input selected from the group consisting of ethylene glycol, glycerol, ethanol methanol or a combination thereof. In addition, the system includes (b) an oxidation biocatalyst including an alcohol oxidase, a copper radical oxidase, a glycerol oxidase, an alditol oxidase or a combination thereof. Further, the system includes (c) an oxidized intermediate. The system also includes (d) a finishing catalyst including a supported metal catalyst, a carboligating catalyst, an amine oxidase, a glyoxalase, an acid catalyst, a base catalyst, an isomerization catalyst or a combination thereof. Still further, the system includes (e) an output.