Nucleic acids, proteins and processes for producing amides
AssigneeLeibniz-Institut Fur Pflanzenbiochemie
InventorsThomas Vogt, Arianne Schnabel, Benedikt Athmer, Kerstin Manke
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Abstract
The invention provides polynucleotides and nucleic acid molecules encoding piperine synthases and proteins and enzyme functional for converting piperoyl-CoA and piperidine to piperine. The invention also provides a method of producing an amide from an acyl-CoA and an amine, and to a method of producing an amide from a carboxylic acid and an amine.
Claims19
A polynucleotide the nucleotide sequence of which is:
- (a) the nucleotide sequence defined in SEQ ID NO:
- 1, or
- (b) a nucleotide sequence of at least 93% sequence identity to SEQ ID NO:
- 1, or
- (c) a nucleotide sequence encoding an amino acid sequence comprising or consisting of the amino acid sequence defined in SEQ ID NO:
- 2, or
- (d) a nucleotide sequence encoding an amino acid sequence of at least 70% sequence identity to the amino acid sequence defined in SEQ ID NO:
- 2, or
- (e) a nucleotide sequence encoding an amino acid sequence of at least 75% sequence similarity to the amino acid sequence defined in SEQ ID NO:
- 2, or
- (f) a nucleotide sequence encoding an amino acid sequence of from 1 to 138 amino acid substitutions, additions, insertions and/or deletions compared to the amino acid sequence defined in SEQ ID NO:
- 2, or
- (g) a fragment of the nucleotide sequence defined in any one of items (a) to (f), the fragment being at most 42 nucleotides shorter due to 5′- and/or 3′-terminal deletions of the nucleotide sequence defined in any one of items (a) to (f), or
- (a′) the nucleotide sequence defined in SEQ ID NO:
- 3, or
- (b′) a nucleotide sequence of at least 93% sequence identity to SEQ ID NO:
- 3, or
- (c′) a nucleotide sequence encoding an amino acid sequence comprising or consisting of the amino acid sequence defined in SEQ ID NO:
- 4, or
- (d′) a nucleotide sequence encoding an amino acid sequence of at least 70% sequence identity to the amino acid sequence defined in SEQ ID NO:
- 4, or
- (e′) a nucleotide sequence encoding an amino acid sequence of at least 75% sequence similarity to the amino acid sequence defined in SEQ ID NO:
- 4, or
- (f′) a nucleotide sequence encoding an amino acid sequence of from 1 to 138 amino acid substitutions, additions, insertions and/or deletions compared to the amino acid sequence defined in SEQ ID NO:
- 4, or
- (g′) a fragment of the nucleotide sequence defined in any one of items (a′) to (f′), the fragment being at most 42 nucleotides shorter due to 5′- and/or 3′-terminal deletions of the nucleotide sequence defined in any one of items (a′) to (f′).
(canceled)
A protein, or protein composition comprising said protein, said protein or protein composition having a catalytic activity for converting piperoyl-CoA and piperidine to piperine with a catalytic activity of at least 0.8, preferably at least 2.4, and most preferably at least 5.0 nkat/mg of said protein or protein composition, respectively, at 30° C. in 30 mM Tris-HCl buffer pH 8.0, containing 1 mM dithiothreitol (DTT), with substrate concentrations of 500 μM piperoyl-CoA and 20 mM piperidine.
A protein comprising a polypeptide the amino acid sequence of which
- (i) is or comprises the amino acid sequence defined in SEQ ID NO:
- 2, or
- (ii) comprises or consists of an amino acid sequence that has at least 70% sequence identity to the amino acid sequence defined in SEQ ID NO:
- 2, or
- (iii) comprises or consists of an amino acid sequence that has at least 75% sequence similarity to the amino acid sequence defined in SEQ ID NO:
- 2, or
- (iv) comprises or consists of an amino acid sequence of from 1 to 138 amino acid substitutions, additions, insertions and/or deletions compared to the amino acid sequence defined in SEQ ID NO:
- 2, or
- (v) is a fragment of the amino acid sequence defined in any one of items (i) to (iv), the fragment being at most 14 amino acid residues shorter due to N- and/or C-terminal deletions of the amino acid sequence defined in any one of items (i) to (iv), or (i′) is or comprises the amino acid sequence defined in SEQ ID NO:
- 4, or
- (ii′) comprises or consists of an amino acid sequence that has at least 70% sequence identity to the amino acid sequence defined in SEQ ID NO:
- 4, or
- (iii′) comprises or consists of an amino acid sequence that has at least 75% sequence similarity to the amino acid sequence defined in SEQ ID NO:
- 4, or
- (iv′) comprises or consists of an amino acid sequence of from 1 to 138 amino acid substitutions, additions, insertions and/or deletions compared to the amino acid sequence defined in SEQ ID NO:
- 4.
(canceled)
Dependencies are inferred from the claim text.