Modified host cells for high efficiency production of vanillin
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.
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.