The present invention provides a nitrilase mutant protein with increased thermal stability and its application in the synthesis of an anti-epileptic drug intermediate, wherein the mutant is obtained by mutating one or two of the amino acids at position 151, 223 and 205 of the amino acid sequence shown in SEQ ID No. 2. the thermal stability of the nitrilase mutant AcN-T151V/C223A/C250G was increased by up to 1.73 folds. The yield of the final product was up to 95% using the recombinant Escherichia coli containing the nitrilase mutant to hydrolyze 1M 1-cyanocyclohexylacetonitrile to produce 1-cyanocyclohexyl acetic acid at 35° C. And the yield of the final product was up to 97% when hydrolyzing 1.2M 1-cyanocyclohexylacetonitrile at 35° C. The final yield was up to 80% when using the nitrilase mutants obtained by the present invention to synthesize gabapentin.

The present application provides engineered polypeptides having imine reductase activity, polynucleotides encoding the engineered imine reductases, host cells capable of expressing the engineered imine reductases, and methods of using these engineered polypeptides with a range of ketone and amine substrate compounds to prepare secondary and tertiary amine product compounds.

Provided is a novel method for producing 4-aminocinnamic acid from 4-nitrophenylalanine. This method comprises: converting 4-nitrophenylalanine into 4-nitrocinnamic acid; and converting 4-nitrocinnamic acid into 4-aminocinnamic acid.

Provided is a novel method for producing 4-aminocinnamic acid from 4-nitrophenylalanine. This method comprises: converting 4-nitrophenylalanine into 4-nitrocinnamic acid; and converting 4-nitrocinnamic acid into 4-aminocinnamic acid.

The present invention provides improved P450-BM3 variants with improved activity. In some embodiments, the P450-BM3 variants exhibit improved activity over a wide range of substrates.

The disclosure relates to engineered ketoreductase polypeptides and processes of using the polypeptides for production of phenylephrine.

Described herein is a method for preparing an amino salt compound, the method including: i) providing a carbonyl compound; ii) reacting the carbonyl compound provided according to (i) in the presence of a transaminase with ii-a) at least one primary amine; and ii-b) at least one carboxylic acid; thereby obtaining a mixture including an at least partially crystallized amino salt compound including a cation and a carboxylate anion based on the at least one carboxylic acid added according to (ii-b). Also described herein is an amino salt compound obtained or obtainable by the method and to the amino salt compound, and a composition including a) an amine of general formula (IIa); and b) at least one carboxylic acid of general formula (III).

Described herein is a method for preparing an amino salt compound, the method including: i) providing a carbonyl compound; ii) reacting the carbonyl compound provided according to (i) in the presence of a transaminase with ii-a) at least one primary amine; and ii-b) at least one carboxylic acid; thereby obtaining a mixture including an at least partially crystallized amino salt compound including a cation and a carboxylate anion based on the at least one carboxylic acid added according to (ii-b). Also described herein is an amino salt compound obtained or obtainable by the method and to the amino salt compound, and a composition including a) an amine of general formula (IIa); and b) at least one carboxylic acid of general formula (III).

A method of producing isothiocyanates includes forming, in a semi-solid or solid callus induction medium, compact callus aggregates from cells obtained from explant material of a Brassica oleracea L. plant. The method includes transferring cells from the callus aggregates, e.g. transferring the callus aggregates, to a suspension culture in a liquid medium in a shake flask, the liquid medium containing a plurality of elicitors. The method further involves transferring, after culturing in the shake flask, cells from the suspension culture to a further suspension culture in a bioreactor containing the elicitors. The method includes extracting and/or purifying of at least one isothiocyanate from cells obtained from the bioreactor. The elicitors have chitosan and salicylic acid to increase accumulation of benzyl isothiocyanate or any other isothiocyanate.

A method of producing isothiocyanates includes forming, in a semi-solid or solid callus induction medium, compact callus aggregates from cells obtained from explant material of a Brassica oleracea L. plant. The method includes transferring cells from the callus aggregates, e.g. transferring the callus aggregates, to a suspension culture in a liquid medium in a shake flask, the liquid medium containing a plurality of elicitors. The method further involves transferring, after culturing in the shake flask, cells from the suspension culture to a further suspension culture in a bioreactor containing the elicitors. The method includes extracting and/or purifying of at least one isothiocyanate from cells obtained from the bioreactor. The elicitors have chitosan and salicylic acid to increase accumulation of benzyl isothiocyanate or any other isothiocyanate.