Methods for the Production of Tryptophan Derivatives
AssigneeNatural MedTech Pty Ltd
InventorsSimon Colin KESSLER, Yit-Heng CHOOI, Mark Heinrich Wolfgang HESTERMANN, Luke Andrew MCFARLANE
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Abstract
The present disclosure relates generally to the biosynthetic production of tryptophan derivatives, in particular tryptamine and tryptamine-derived alkaloids.
Claims69
A cell capable of producing at least one tryptophan derivative comprising at least one exogenous polynucleotide encoding a pyridoxal phosphate (PLP)-dependent tryptophan decarboxylase,
- wherein the level of production of the at least one tryptophan derivative in the cell is increased relative to the level of production of a second tryptophan derivative in an equivalent cell lacking the exogenous polynucleotide or comprising a second polynucleotide encoding a second tryptophan decarboxylase, and wherein the second polynucleotide is not the same as the exogenous polynucleotide.
A cell capable of producing at least one tryptophan derivative comprising at least one exogenous polynucleotide encoding a pyridoxal phosphate (PLP)-dependent tryptophan decarboxylase, wherein the level of production of the at least one tryptophan derivative in the cell is increased relative to the level of production of a second tryptophan derivative in an equivalent cell lacking the exogenous polynucleotide or comprising a second polynucleotide encoding a second tryptophan decarboxylase, and wherein the second polynucleotide is not the same as the exogenous polynucleotide and wherein the PLP-dependent tryptophan decarboxylase comprises one or more or all of:
- a) a first amino acid sequence of GX1X2X3X4X5X6X7X8X9X10X11X12X13X14X15X16X17R wherein:
- X1, X2, X3, X4, X8, X9, X10, X11, X12, X13, X14, X15, and X16 are any amino acid residue,
- X5 and X6 are an amino acid residue selected from the group consisting of glycine, alanine, serine and threonine,
- X7 is an amino acid residue selected from the group consisting of serine and threonine, and
- X17 is an amino acid residue selected from the group consisting of alanine and glycine;
- b) a second amino acid sequence of X1X2X3X4X5X6HX7X8X9X10KX11X12X13X14X15X16 wherein:
- X1 is any amino acid residue selected from the group consisting of tyrosine, leucine, phenylalanine, valine, cysteine and tryptophan,
- X2, X4, X5, X6, X7, X8, X9, X10, X11, X12, X13, X14 and X15 are any amino acid residue,
- X3 is any amino acid residue selected from the group consisting of serine and threonine, and
- X16 is an amino acid residue selected from the group consisting of glycine, alanine, serine and threonine;
- c) a third amino acid sequence of DX1X2X3X4X5X6X7X8X9X10X11X12X13X14GX15X16X17X18X19X20X21D wherein:
- X1, X2, X3, X4, X5, X6, X8, X9, X10, X11, X12, X13, X14, X15, X17, X18, X19 and X21 are any amino acid residue, and
- X7 is an amino acid residue selected from the group consisting of proline and asparagine,
- X16 is an amino acid residue selected from the group consisting of threonine, proline and valine, and
- X20 is 1, 2, 3, 4, 5, or 6 amino acid residues and is any amino acid residue;
- d) a fourth amino acid sequence of X1X2HX3X4X5X6X7X8X9X10 wherein:
- X1 is an amino acid residue selected from the group consisting of tryptophan and phenylalanine,
- X2, X3, X6, X9 and X10 are any amino acid residue,
- X4 is an amino acid residue selected from the group consisting of aspartic acid and glutamic acid,
- X5 is an amino acid residue selected from the group consisting of glycine, alanine and serine,
- X7 is an amino acid residue selected from the group consisting of phenylalanine, tyrosine and tryptophan, and
- X8 is an amino acid residue selected from the group consisting of glycine, alanine, serine and threonine;
- and
- e) a fifth amino acid sequence of X1X2X3X4X5X6X7HKX8X9X10X11X12X13X14X15X16 wherein:
- X1 is an amino acid residue selected from the group consisting of aspartic acid, glutamic acid, histidine and asparagine,
- X2 is an amino acid residue selected from the group consisting of serine and threonine,
- X3, X4, X5, X7, X10, X11, X12, X13 and X14 are any amino acid residue,
- X6 is an amino acid residue selected from the group consisting of aspartic acid, glutamic acid, asparagine, serine and threonine,
- X8 is an amino acid residue selected from the group consisting of tryptophan, phenylalanine, tyrosine, leucine, methionine and cysteine,
- X9 is an amino acid residue selected from the group consisting of leucine, isoleucine, valine, methionine and phenylalanine,
- X15 is an amino acid residue selected from the group consisting of cysteine, alanine and serine, and
- X16 is an amino acid residue selected from the group consisting of glycine, serine, alanine and threonine.
A cell capable of producing at least one tryptophan derivative comprising at least one exogenous polynucleotide encoding an amino acid sequence set forth in any one of SEQ ID NOs:1, 3, 5, 7, 29 and 48 or a sequence having at least about 70% sequence identity thereto,
- wherein the level of production of the at least one tryptophan derivative in the cell is increased relative to the level of production of a second tryptophan derivative in an equivalent cell lacking the exogenous polynucleotide or comprising a second polynucleotide encoding a second tryptophan decarboxylase, and wherein the second polynucleotide is not the same as the exogenous polynucleotide.
A cell capable of producing at least one tryptophan derivative, comprising:
- a) at least one exogenous polynucleotide encoding a PLP-dependent tryptophan decarboxylase having an amino acid sequence set forth in any one of SEQ ID NOs:
- 1, 3, 5, 7, 29 and 48, or a sequence having at least about 70% sequence identity thereto;
- b) at least one exogenous polynucleotide encoding a PsiH tryptamine 4-monooxygenase;
- c) at least one exogenous polynucleotide encoding a PsiK hydroxytryptamine kinase;
- and
- d) at least one exogenous polynucleotide encoding a PsiM methyl transferase.
The cell of claim 48 or 49, wherein the cell is capable of an increased level of production of the at least one tryptophan derivative relative to the level of production of at least one tryptophan derivative in an equivalent cell comprising a second polynucleotide encoding a second tryptophan decarboxylase, wherein the second polynucleotide is not the same as (a), and optionally wherein the second polynucleotide encodes a PsiD tryptophan decarboxylase.
A vector comprising a polynucleotide encoding a PLP-dependent tryptophan decarboxylase having an amino acid sequence set forth in any one of SEQ ID NOs:
- 1, 3, 5, 7, 29 and 48, optionally wherein the polynucleotide is operably linked to a heterologous promoter capable of directing expression of the polynucleotide in a cell that does not naturally express said polynucleotide.
A method for producing at least one tryptophan derivative, the method comprising contacting tryptophan with at least one pyridoxal phosphate (PLP)-dependent tryptophan decarboxylase comprising an amino acid sequence set forth in any one of SEQ ID NOs:1, 3, 5, 7, 29 and 48 or a sequence having at least about 70% sequence identity thereto, under conditions suitable for the production of the at least one tryptophan derivative.
A composition comprising the cell of any one of claims 1 to 54 and 57, or the extracted tryptophan derivative of claim 67.
Dependencies are inferred from the claim text.