Production of vanillin
AssigneePlant Bioscience Limited
InventorsArjan Narbad, Michael John Charles Rhodes, Michael John Gasson, Nicholas John Walton
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Abstract
A method of producing vanillin comprising the steps of: (1) providing trans-ferulic acid or a salt thereof; and (2) providing trans-ferulate: CoASH ligase activity (enzyme activity I), trans-feruloyl ScoA hydratase activity (enzyme activity II), and 4-hydroxy-3-methoxyphenyl-β-hydroxy-propionyl SCoA (HMPHP SCoA) cleavage activity (enzyme activity III). Conveniently the enzymes are provided by Pseudomonas fluorescens Fe3 or a mutant or derivative thereof. Polypeptides with enzymes activities II and III and polynucleotides encoding the polypeptides. Use of the polypeptides or the polynucleotides in a method for producing vanillin is also provided.
Claims16
An isolated polynucleotide contained within the cosmid clone pFI 793 as deposited under the Budapest Treaty at the National Collections of Industrial and Marine Bacteria Limited, Scotland under Accession No NCIMB 40777, the isolated polynucleotide hybridizing to the complement of SEQ ID NO:
- 1 when incubated in an aqueous solution containing 2×SSC and at a temperature of 65° C.
Cosmid pFI 793 as deposited under the Budapest Treaty at the National Collections of Industrial and Marine Bacteria Limited, Scotland under Accession No NCIMB 40777.
An isolated polynucleotide that hybridizes to the complement of SEQ ID NO:
- 1, when incubated in an aqueous solution containing 2×SSC and at a temperature of 65° C., and encodes a polypeptide which, in the presence of appropriate cofactors if any, is capable of catalyzing the conversion of trans-feruloyl-S-CoA to vanillin via 4-hydroxy-3-methoxyphenyl-β-hydroxy-propionyl SCoA (HMPHP SCoA).
An isolated polynucleotide comprising a nucleotide sequence of SEQ ID NO:
- 1 or fragments thereof encoding enzymatically active polypeptides, which fragments hybridize to the complement of SEQ ID NO:
- 1 when incubated in an aqueous solution containing 2×SSC and at a temperature of 65° C.
A transgenic plant comprising at least one polynucleotide that hybridizes to the complement of SEQ ID NO:
- 1 when incubated in an aqueous solution containing 2×SSC and at a temperature of 65° C., wherein said plant, as a consequence of the presence of said polynucleotide, expresses trans-feruloyl ScoA hydratase activity and 4-hydroxy-3-methoxyphenyl-β-hydroxy-propionyl ScoA cleavage activity.
Dependencies are inferred from the claim text.