Modified host cells for high efficiency production of vanillin

Patent metadata

Field	Value
Publication number	US-12480101-B2
Application number	US-202017631429-A
Country	US
Kind code	B2
Filing date	Jul 31, 2020
Priority date	Aug 1, 2019
Publication date	Nov 25, 2025
Grant date	Nov 25, 2025

Abstract

Provided herein are genetically modified host cells, compositions, and methods for improved production of vanillin and/or glucovanillin. The host cells, compositions, and methods described herein provide an efficient route for the heterologous production of vanillin and/or glucovanillin and any compound that can be synthesized or biosynthesized from either or both.

First claim

What is claimed:

1 . A genetically modified Saccharomyces cerevisiae host cell that produces vanillin or glucovanillin, the host cell comprising:
(i) deletion of a homolog of fatty aldehyde dehydrogenase (HFD1) gene; and (ii) one or more nucleic acids expressing:
a catechol-O-methyltransferase (COMT);
an aromatic carboxylic acid reductase (ACAR) from Nocardia iowensis;
a phosphopantetheinyl transferase (PPTASE); and
a Podospora pauciseta 3-dehydroshikimate dehydratase (AroZ),
wherein the host cell does not express an Hfd1 protein.

2 . The genetically modified Saccharomyces cerevisiae host cell of claim 1 , further comprising deletion of an ADH6 gene.

3 . The genetically modified Saccharomyces cerevisiae host cell of claim 1 , further comprising deletion of a GRE2 gene.

4 . The genetically modified Saccharomyces cerevisiae host cell of claim 1 , further comprising deletion of a YGL039W gene.

5 . The genetically modified Saccharomyces cerevisiae host cell of claim 1 , further comprising one or more nucleic acids expressing a 3-dehydroquinate synthase (AroB) or a dehydroquinate dehydratase (AroD).

6 . The genetically modified Saccharomyces cerevisiae host cell of claim 1 , further comprising one or more nucleic acids expressing an AroB, an AroD, or a phospho-2-dehydro-3-deoxyheptonate aldolase (AroF).

7 . The genetically modified Saccharomyces cerevisiae host cell of claim 1 , further comprising one or more nucleic acids expressing an E. coli AroB, an E. coli AroD, and an E. coli AroF.

8 . The genetically modified Saccharomyces cerevisiae host cell of claim 1 , wherein the PPTASE is a Corynebacterium glutamicum PPTASE.

9 . The genetically modified Saccharomyces cerevisiae host cell of claim 1 , further comprising one or more nucleic acids expressing an eugenol alcohol oxidase (EAO).

10 . The genetically modified Saccharomyces cerevisiae host cell of claim 1 , further comprising one or more nucleic acids expressing a Rhodococcus jostii EAO.

11 . The genetically modified Saccharomyces cerevisiae host cell of claim 1 , further comprising one or more nucleic acids expressing an Arabidopsis thaliana UDP-glycosyltransferase (UGT).

12 . The genetically modified Saccharomyces cerevisiae host cell of claim 1 , wherein the one or more nucleic acids are expressed from an inducible promoter.

13 . The genetically modified Saccharomyces cerevisiae host cell of claim 12 , wherein the inducible promoter is a GAL promoter.

14 . The genetically modified Saccharomyces cerevisiae host cell of claim 12 , wherein the one or more nucleic acids are expressed from a GAL promoter, and wherein the genetically modified Saccharomyces cerevisiae host cell comprises a yeast GAL80 gene expressed from a maltose-responsive promoter.

15 . A method for producing vanillin or glucovanillin, comprising the steps of:
a. culturing the genetically modified Saccharomyces cerevisiae host cell of claim 1 in a medium with a carbon source under conditions suitable for making vanillin or glucovanillin; and b. recovering said vanillin or glucovanillin from the medium.

Assignees

• Amyris Inc

Inventors

• Raetz Lauren
• Hansen Chad

Classifications

• C12Y102/01003

  Aldehyde dehydrogenase (NAD+) (1.2.1.3) · CPC title

• C12P7/24

  containing a carbonyl group · CPC title

• C07C47/58

  Vanillin · CPC title

• C12P19/46

  having an oxygen atom of the saccharide radical bound to a cyclohexyl radical, e.g. kasugamycin · CPC title

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title