A method of producing 2,4-dihydroxybutyric acid (2,4-DHB) by a synthetic pathway that includes transforming malate into 4-phospho-malate using a malate kinase, then transforming 4-phospho-malate into malate-4-semialdehyde using a malate semialdehyde dehydrogenase, and then transforming malate-4-semialdehyde into 2,4-DHB using a DHB dehydrogenase.

A method of producing 2,4-dihydroxybutyric acid (2,4-DHB) by a synthetic pathway that includes transforming malate into 4-phospho-malate using a malate kinase, then transforming 4-phospho-malate into malate-4-semialdehyde using a malate semialdehyde dehydrogenase, and then transforming malate-4-semialdehyde into 2,4-DHB using a DHB dehydrogenase.

Methods for the production of L-glufosinate (also known as phosphinothricin or (S)-2-amino-4-(hydroxy(methyl)phosphonoyl)butanoic acid) are provided. The methods comprise a two-step process. The first step involves the oxidative deamination of D-glufosinate to PPO (2-oxo-4-(hydroxy(methyl)phosphinoyl)butyric acid). The second step involves the specific amination of PPO to L-glufosinate, using an amine group from one or more amine donors. By combining these two reactions, the proportion of L-glufosinate in a mixture of L-glufosinate and D-glufosinate can be substantially increased.

Methods for the production of L-glufosinate (also known as phosphinothricin or (S)-2-amino-4-(hydroxy(methyl)phosphonoyl)butanoic acid) are provided. The methods comprise a two-step process. The first step involves the oxidative deamination of D-glufosinate to PPO (2-oxo-4-(hydroxy(methyl)phosphinoyl)butyric acid). The second step involves the specific amination of PPO to L-glufosinate, using an amine group from one or more amine donors. By combining these two reactions, the proportion of L-glufosinate in a mixture of L-glufosinate and D-glufosinate can be substantially increased.

The present invention is in the technical field of synthetic biology and metabolic engineering. More particularly, the present invention is in the technical field of metabolically engineered cells of microorganisms and the use of said cells in a fermentation. The present invention describes a metabolically engineered cell of a microorganism and a method by fermentation with said cell for production of a sialylated di- and/or oligosaccharide. The metabolically engineered cell comprises a pathway for production of said sialylated di- and/or oligosaccharide, synthesizes sialic acid, expresses at least one sialyltransferase, preferably is modified in the expression or activity of at least one sialyltransferase, is modified to have a fully or partially knocked out or rendered less functional sialic acid catabolic pathway and is modified for overexpression of an endogenous sialic acid transporter and/or expression, preferably overexpression, of an exogenous, homologous and/or heterologous sialic acid transporter. Furthermore, the present invention provides for purification of said sialylated di- and/or oligosaccharide from the cultivation, preferably fermentation.

The present invention is in the technical field of synthetic biology and metabolic engineering. More particularly, the present invention is in the technical field of metabolically engineered cells of microorganisms and the use of said cells in a fermentation. The present invention describes a metabolically engineered cell of a microorganism and a method by fermentation with said cell for production of a sialylated di- and/or oligosaccharide. The metabolically engineered cell comprises a pathway for production of said sialylated di- and/or oligosaccharide, synthesizes sialic acid, expresses at least one sialyltransferase, preferably is modified in the expression or activity of at least one sialyltransferase, is modified to have a fully or partially knocked out or rendered less functional sialic acid catabolic pathway and is modified for overexpression of an endogenous sialic acid transporter and/or expression, preferably overexpression, of an exogenous, homologous and/or heterologous sialic acid transporter. Furthermore, the present invention provides for purification of said sialylated di- and/or oligosaccharide from the cultivation, preferably fermentation.

Geranyl pyrophosphate (GPP) is a key intermediate molecule in the bioproduction of thousands of natural products. Currently, natural products are either cultivated from plants, synthesized via complex chemical synthesis strategies, or through cell-based factories also known as biofoundries. However, in order to replicate the process in a cell free environment, numerous enzymes and cofactors must be utilized making this approach costly and unviable. In order to make this process viable, a new approach was needed that uses fewer enzymes and co-factors. As described herein, the present invention demonstrates that it is possible to create GPP from glycerol through a short and concise biosynthetic pathway outside of the cell.

Geranyl pyrophosphate (GPP) is a key intermediate molecule in the bioproduction of thousands of natural products. Currently, natural products are either cultivated from plants, synthesized via complex chemical synthesis strategies, or through cell-based factories also known as biofoundries. However, in order to replicate the process in a cell free environment, numerous enzymes and cofactors must be utilized making this approach costly and unviable. In order to make this process viable, a new approach was needed that uses fewer enzymes and co-factors. As described herein, the present invention demonstrates that it is possible to create GPP from glycerol through a short and concise biosynthetic pathway outside of the cell.

Disclosed herein are methods for the preparation of boronate derivatives in the synthesis of antimicrobial compounds and uses thereof. Disclosed herein includes method of making a compound of Formula (B) by reducing the ketone group of the keto-ester compound of Formula (A), and the reduction can be performed using a Ruthenium based catalyst system or using an alcohol dehydrogenase bioreduction system.

Disclosed herein are methods for the preparation of boronate derivatives in the synthesis of antimicrobial compounds and uses thereof. Disclosed herein includes method of making a compound of Formula (B) by reducing the ketone group of the keto-ester compound of Formula (A), and the reduction can be performed using a Ruthenium based catalyst system or using an alcohol dehydrogenase bioreduction system.