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US 10,006,060 B2Granted
Selectivity of the production of vanilloids in a recombinant unicellular host
AssigneeRhodia Operations
InventorsOdile Ramaen, Rudy Pandjaitan, Mirjana Gelo-Pujic
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Abstract
The present invention relates to methods for producing vanilloid compounds in a recombinant host, and in particular for converting a protocatechuic aldehyde into a substantially pure vanilloid. It further relates to novel yeast strains that are suitable for producing such vanilloid compounds.
Claims23
A method for producing a substantially pure vanilloid of formula (I):
- R1 being selected from the group consisting of —CHO;
- —COOH;
- —COOR3;
- —CH2OH;
- —CH2COOH;
- —C(═O)CH3;
- —CR3(OH)COOH;
- —CHR3COOH;
- —CH2NHC(═O)R3;
- wherein R3 is a lower alkyl,
- R2 being different from a methyl (—CH3),
- comprising the steps of:
- a) providing a recombinant unicellular host capable of producing said vanilloid, wherein said recombinant host expresses at least a nucleic acid coding for a 3-dehydroshikimate dehydratase (3DSD) and at least a nucleic acid coding for an aromatic carboxylic acid reductase (ACAR);
- b) cultivating said host in a suitable medium;
- and
- c) recovering the produced vanilloid from said host or from the culture supernatant thereof,
- wherein said recombinant host expresses at least a nucleic acid coding for a caffeic acid 3-O-methyltransferase polypeptide that is suitable for methylating selectively the 3-OH of a protocatechuic aldehyde.
14 more elements
A yeast suitable for producing a substantially pure vanilloid of formula (I):
- R1 being selected from the group consisting of —CHO;
- —COOH;
- —COOR3;
- —CH2OH;
- —CH2COOH;
- —C(═O)CH3;
- —CR3(OH)COOH;
- —CHR3COOH;
- —CH2NHC(═O)R3;
- wherein R3 is a lower alkyl,
- R2 being different from a methyl (—CH3),
- expressing at least a nucleic acid coding for an aromatic carboxylic acid reductase (ACAR) and a 3-dehydroshikimate dehydratase (3DSD), wherein said yeast expresses at least a nucleic acid coding for a caffeic acid 3-O-methyltransferase polypeptide that is suitable for methylating selectively the 3-OH of a protocatechuic aldehyde.
9 more elements
A method for converting a protocatechuic aldehyde into a substantially pure vanilloid of formula (I):
- R1 being selected from the group consisting of —CHO;
- —COOH;
- —COOR3;
- —CH2OH;
- —CH2COOH;
- —C(═O)CH3;
- —CR3(OH)COOH;
- —CHR3COOH;
- —CH2NHC(═O)R3;
- wherein R3 is a lower alkyl,
- R2 being different from a methyl (—CH3),
- and comprising the steps of:
- a) providing a recombinant unicellular host capable of producing said vanilloid, wherein said recombinant host expresses at least a nucleic acid coding for an aromatic carboxylic acid reductase (ACAR) and does not express nucleic acids coding for the following enzymes:
- a phenylalanine ammonia lyase (PAL) or a tyrosine ammonia lyase (TAL) or a phenylalanine/tyrosine ammonia lyase (PAL/TAL), a coA ligase, and a crotonase;
- b) cultivating said host in a suitable medium;
- and
- c) recovering the produced vanilloid from said host or from the culture supernatant thereof,
- wherein said recombinant host expresses at least a nucleic acid coding for a caffeic acid 3-O-methyltransferase polypeptide that is suitable for methylating selectively the 3-OH of a protocatechuic aldehyde.
15 more elements
A method for converting a protocatechuic aldehyde into a substantially pure vanilloid of formula (I):
- R1 being selected from the group consisting of —CHO;
- —COOH;
- —COOR3;
- —CH2OH;
- —CH2COOH;
- —C(═O)CH3;
- —CR3(OH)COOH;
- —CHR3COOH;
- —CH2NHC(═O)R3;
- wherein R3 is a lower alkyl,
- R2 being different from a methyl (—CH3),
- and comprising the steps of:
- a) providing a recombinant unicellular host capable of producing said vanilloid, wherein said recombinant host expresses at least a nucleic acid coding for an aromatic carboxylic acid reductase (ACAR) and at least one nucleic acid coding for a 3-dehydroshikimate dehydratase (3DSD);
- b) cultivating said host in a suitable medium;
- and
- c) recovering the produced vanilloid from said host or from the culture supernatant thereof,
- wherein said recombinant host expresses at least a nucleic acid coding for a caffeic acid 3-O-methyltransferase polypeptide that is suitable for methylating selectively the 3-OH of a protocatechuic aldehyde.
14 more elements
A yeast suitable for converting a protocatechuic aldehyde into a substantially pure vanilloid of formula (I):
- R1 being selected from the group consisting of —CHO;
- —COOH;
- —COOR3;
- —CH2OH;
- —CH2COOH;
- —C(═O)CH3;
- —CR3(OH)COOH;
- —CHR3COOH;
- —CH2NHC(═O)R3;
- wherein R3 is a lower alkyl,
- R2 being different from a methyl (—CH3),
- expressing at least a nucleic acid coding for an aromatic carboxylic acid reductase (ACAR), wherein said yeast expresses at least a nucleic acid coding for a caffeic acid 3-O-methyltransferase polypeptide that is suitable for methylating selectively the 3-OH of a protocatechuic aldehyde, and does not express nucleic acids coding for the following enzymes:
- a phenylalanine ammonia lyase (PAL) or a tyrosine ammonia lyase (TAL) or a phenylalanine/tyrosine ammonia lyase (PAL/TAL), a coA ligase, and a crotonase.
10 more elements
A yeast suitable for converting a protocatechuic aldehyde into a substantially pure vanilloid of formula (I):
- R1 being selected from the group consisting of —CHO;
- —COOH;
- —COOR3;
- —CH2OH;
- —CH2COOH;
- —C(═O)CH3;
- —CR3(OH)COOH;
- —CHR3COOH;
- —CH2NHC(═O)R3;
- wherein R3 is a lower alkyl,
- R2 being different from a methyl (—CH3),
- expressing at least a nucleic acid coding for an aromatic carboxylic acid reductase (ACAR) and at least a nucleic acid coding for a 3-dehydroshikimate dehydratase (3DSD), wherein said yeast expresses at least a nucleic acid coding for a caffeic acid 3-O-methyltransferase polypeptide that is suitable for methylating selectively the 3-OH of a protocatechuic aldehyde.
9 more elements
Dependencies are inferred from the claim text.