Vanillin Synthase
AssigneesEvolva SA, University of Copenhagen
InventorsBirger Lindberg Møller, Esben Halkjaer Hansen, Jørgen Hansen, Nethaji Janeshawari Gallage
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Abstract
The invention relates to methods for producing vanillin and related compounds. The methods involve use of a vanillin synthase capable of catalyzing side chain cleavage of ferulic acid to form vanillin. The invention also relates to host organisms expressing such vanillin synthases useful in the methods.
Claims42
A method of producing vanillin, vanillyl alcohol, vanillin glucoside and/or vanillyl alcohol glucoside, said method comprising
- a) providing a microbial organism, wherein said microbial organism
- i. is capable of producing ferulic acid and/or ferulic acid glucoside;
- and
- ii. comprises a heterologous nucleic acid encoding a vanillin synthase, wherein said vanillin synthase is an enzyme capable of catalyzing side chain cleavage of ferulic acid to form vanillin;
- and
- b) cultivating said microbial organism in culture medium supporting growth of said microbial organism
- c) Isolating vanillin, vanillyl alcohol, vanillyl alcohol glucoside and/or vanillin glucoside from the microbial organism and/or from the culture medium.
A microbial organism wherein said microbial organism
- i. is capable of producing ferulic acid;
- and
- ii. comprises a heterologous nucleic acid encoding vanillin synthase, wherein said vanillin synthase is an enzyme capable of catalyzing side chain cleavage of ferulic acid to form vanillin.
The method or the microbial organism according to any one of the preceding claims, wherein the microbial organism is selected from the group consisting of bacteria and fungi.
The method or the microbial organism according to any one of the preceding claims, wherein the microbial organism is a yeast.
The method or the microbial organism according to any one of the preceding claims, wherein the microbial organism comprises at least one heterologous nucleic acid encoding an enzyme involved in synthesis of ferulic acid.
The method according to any one of claims or the cell according to any one of claims, wherein the enzyme involved in synthesis of ferulic acid is Vanillhyl alcohol oxidase.
A method of producing vanillin said method comprising the steps of
- a) providing a microbial organism, wherein said microbial organism comprises a heterologous nucleic acid encoding vanillin synthase, wherein said vanillin synthase is an enzyme capable of catalyzing side chain cleavage of ferulic acid to form vanillin;
- and
- b) cultivating said microbial organism in the presence of ferulic acid and/or a ferulic acid derivative in culture medium supporting growth of said microbial organism;
- and
- c) Isolating vanillin, vanillyl alcohol, vanillyl alcohol glucoside and/or vanillin glucoside from the microbial organism and/or from the culture medium.
A method for producing vanillin and/or a vanillin derivative, said method comprising
- a) providing ferulic acid and/or a ferulic acid derivative
- b) contacting said ferulic acid and/or ferulic acid derivative with vanillin synthase, wherein said vanillin synthase is an enzyme capable of catalyzing side chain cleavage of ferulic acid to form vanillin thereby producing vanillin and/or a vanillin derivative.
The method according to claim 18, wherein the method is performed in vitro.
The method according to any one of claims, wherein the ferulic acid derivative is ferulic acid glucoside and the vanillin derivative is vanillin glucoside.
A method of producing vanillin, vanillyl alcohol, vanillyl alcohol glucoside and/or vanillin glucoside, said method comprising
- a) Providing a plant comprising a heterologous nucleic acid encoding vanillin synthase, wherein said vanillin synthase is an enzyme capable of catalyzing side chain cleavage of ferulic acid to form vanillin;
- and
- b) cultivating said plant;
- and
- c) Isolating vanillin from the plant.
A plant comprising a heterologous nucleic acid encoding vanillin synthase, wherein said vanillin synthase is an enzyme capable of catalyzing side chain cleavage of ferulic acid to form vanillin and a heterologous encoding a glucosyl transferase.
A method of producing animal feed, said method comprising
- a) Providing a plant comprising a heterologous nucleic acid encoding vanillin synthase, wherein said vanillin synthase is an enzyme capable of catalyzing side chain cleavage of ferulic acid to form vanillin;
- and
- b) cultivating said plant;
- and
- c) processing the plant into animal feed.
A method of producing food product, said method comprising
- a) Providing a plant comprising edible parts, wherein said plant comprises a heterologous nucleic acid encoding vanillin synthase, wherein said vanillin synthase is an enzyme capable of catalyzing side chain cleavage of ferulic acid to form vanillin;
- and
- b) cultivating said plant;
- and
- c) harvesting said edible parts;
- thereby obtaining a food product with vanillin palate.
The method, the microbial organism or the plant according to any one of the preceding claims, wherein the vanillin synthase is vanillin synthase of SEQ ID NO:1 or a functional homologue thereof sharing at least 70%, such as at least 80%, for example at least 85%, such as at least 90%, for example at least 95% sequence identity therewith.
The method, the plant or the microbial organism according to any one of the preceding claims, wherein the microbial organism or the plant also comprises a heterologous nucleic acid encoding a glucosyl transferase.
Dependencies are inferred from the claim text.