The present disclosure provides recombinant microorganisms and methods for the production of 4-HMF, 2,4-furandimethanol, furan-2,4-dicarbaldehyde, 4-(hydroxymethyl)furoic acid, 2-formylfuran-4-carboxylate, 4-formylfuran-2-carboxylate, and/or 2,4-FDCA from a carbon source. The method provides for engineered microorganisms that express endogenous and/or exogenous nucleic acid molecules that catalyze the conversion of a carbon source into 4-HMF, 2,4-furandimethanol, furan-2,4-dicarbaldehyde, 4-(hydroxymethyl)furoic acid, 2-formylfuran-4-carboxylate, 4-formylfuran-2-carboxylate, and/or 2,4-FDCA. The disclosure further provides methods of producing polymers derived from 4-IMF, 2,4-furandimethanol, furan-2,4-dicarbaldehyde, 4-(hydroxymethyl)furoic acid, 2-formylfuran-4-carboxylate, 4-formylfuran-2-carboxylate, and/or 2,4-FDCA.

The present disclosure provides cytochrome P450 variants useful for carrying out in vivo and in vitro carbene insertion reactions. Methods for preparing carbene insertion products including cyclopropenes, cyclopropanes, bicyclobutanes, substituted lactones, cyclized compounds, and substituted amines are also described.

The present disclosure provides cytochrome P450 variants useful for carrying out in vivo and in vitro carbene insertion reactions. Methods for preparing carbene insertion products including cyclopropenes, cyclopropanes, bicyclobutanes, substituted lactones, cyclized compounds, and substituted amines are also described.

The present disclosure includes methods and systems for removing one or more fermentation inhibitors (e.g., furfural) from a hydrolysate composition (e.g., a whole broth hydrolysate) by gas stripping the one or more fermentation inhibitors from the hydrolysate composition.

The present disclosure includes methods and systems for removing one or more fermentation inhibitors (e.g., furfural) from a hydrolysate composition (e.g., a whole broth hydrolysate) by gas stripping the one or more fermentation inhibitors from the hydrolysate composition.

Method for preparing a 2,5-furandicarboxylic acid (“FDCA”) comprising: contacting a polypeptide comprising an amino acid sequence that has greater than 34% sequence identity with an amino acid sequence set out in SEQ ID NO:1 or SEQ ID NO:2 with furoic acid in the presence of carbon dioxide; wherein the polypeptide has carboxylase and decarboxylase activity and comprises (i) the amino acid corresponding to H297 or a functional substitution thereof and (ii) at least one of (a) the amino acid corresponding to R305 or a functional substitution thereof; and (b) the amino acid corresponding to R332 or a functional substitution thereof; wherein the position is numbered relative to the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2.

Method for preparing a 2,5-furandicarboxylic acid (“FDCA”) comprising: contacting a polypeptide comprising an amino acid sequence that has greater than 34% sequence identity with an amino acid sequence set out in SEQ ID NO:1 or SEQ ID NO:2 with furoic acid in the presence of carbon dioxide; wherein the polypeptide has carboxylase and decarboxylase activity and comprises (i) the amino acid corresponding to H297 or a functional substitution thereof and (ii) at least one of (a) the amino acid corresponding to R305 or a functional substitution thereof; and (b) the amino acid corresponding to R332 or a functional substitution thereof; wherein the position is numbered relative to the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2.

The present invention provides a Comamonas testosteroni strain and use thereof for the synthesis of 5-hydroxymethyl-2-furancarboxylic acid, wherein the strain is Comamonas testosteroni SC1588, maintained in China Center for Type Culture Collection, CCTCC for short, on Oct. 13, 2016, with an accession number of CCTCC NO: M 2016562.

The present invention provides a Comamonas testosteroni strain and use thereof for the synthesis of 5-hydroxymethyl-2-furancarboxylic acid, wherein the strain is Comamonas testosteroni SC1588, maintained in China Center for Type Culture Collection, CCTCC for short, on Oct. 13, 2016, with an accession number of CCTCC NO: M 2016562.

The present application relates to enantioselective enzymatic reduction of keto compounds to the corresponding chiral hydroxy compounds. Specifically the present application describes enantioselective enzymatic reduction of ethyl-4-chloroacetoacetate (compound of formula I) into (R)-ethyl-4-chloro-3-hydroxybutyrate (compound of formula II). The present application also covers use of (R)-ethyl-4-chloro-3-hydroxybutyrate prepared by the enantioselective enzymatic reduction process in the preparation of SGLT2 inhibitor empaglifiozin.

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