Pterin-dependent biocatalysts and uses thereof
Provided herein are biocatalysts and systems thereof for pterin-dependent enzymes and pathways and methods of making and using the same. Provided herein in some embodiments are biocatalysts having a pterin source and a pterin-dependent enzymatic pathway biologically coupled to the pterin source. Tetrahydrobiopterin (referred to herein as BH4 or BH 4) can be the pterin source. The BH4 can be synthesized by a tetrahydrobiopterin synthesis pathway. The tetrahydrobiopterin synthesis pathway can include a GTP cyclohydrase; a pyruvoyl tetrahydropterin synthase; a sepiapterin reductase, and/or any combination thereof. The biocatalyst can further contain a pterin-dependent enzymatic pathway. The pterin-dependent enzymatic pathway can be amino acid mono-oxygenase, phenylalanine hydroxylase, tryptophan hydroxylase, tyrosine hydroxylase, nitric oxide synthase, alkylglycerol monooxygenase, and/or any combination thereof.
1. A method of biocatalysis comprising the steps of:
providing a metabolite of lignin to a biocatalyst comprising a pterin-dependent enzymatic pathway in the presence of a tetrahydrobiopterin source, wherein the pterin-dependent enzymatic pathway comprises an amino acid mono-oxygenase, and
wherein the metabolite of lignin is one of:
and
wherein the amino acid mono-oxygenase is encoded by a nucleotide sequence comprising SEQ ID NO: 14 or SEQ ID NO: 15.
2. The method of claim 1 , wherein the biocatalyst further comprises a tetrahydrobiopterin recycling pathway, the tetrahydrobiopterin recycling pathway comprising:
a pterin-4a-carbinolamine dehydratase; and
a dihydropterin reductase.
3. The method of claim 1 , wherein the metabolite of lignin is:
4. The method of claim 1 , wherein the tetrahydrobiopterin source is a tetrahydrobiopterin synthesis pathway comprising:
a GTP cyclohydrolase;
a pyruvoyl tetrahydropterin synthase; and
a sepiapterin reductase.
5. The method of claim 1 , wherein the biocatalyst further comprises an aromatic-l-amino acid decarboxylase.
6. The method of claim 1 , wherein the biocatalyst further comprises a terpene alkaloid synthase.
7. A method of biocatalysis comprising the steps of:
providing a metabolite of lignin to a biocatalyst comprising a pterin-dependent enzymatic pathway in the presence of a tetrahydrobiopterin source, wherein the pterin-dependent enzymatic pathway comprises an amino acid mono-oxygenase, and
wherein the metabolite of lignin is:
8. The method of claim 2 , wherein the pterin-4a-carbinolamine dehydratase is encoded by a nucleotide sequence comprising SEQ ID NO: 12; and
the dihydropterin reductase is encoded by a nucleotide sequence comprising SEQ ID NO: 13.
9. The method of claim 4 , wherein the GTP cyclohydrolase is encoded by a nucleotide sequence comprising SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4;
the pyruvoyl tetrahydropterin synthase is encoded by a nucleotide sequence comprising SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, or SEQ ID NO: 8; and
the sepiapterin reductase is encoded by a nucleotide sequence comprising SEQ ID NO: 9, SEQ ID NO: 10, or SEQ ID NO: 11.
10. The method of claim 5 , wherein the aromatic-l-amino acid decarboxylase is encoded by a nucleotide sequence comprising SEQ ID NO: 16.
11. The method of claim 6 , wherein the terpene alkaloid synthase is encoded by a nucleotide sequence comprising SEQ ID NO: 17.