GENETICALLY MODIFIED PROBIOTIC FOR THE TREATMENT OF PHENYLKETONURIA (PKU) DISEASE
A GMP adapted to provide the PAL gene for the treatment of PKU when administered orally. The GMP of the present invention may include a probiotic, a PAL gene to be expressed using the probiotic, wherein the PAL gene is functionally attached to a promoter and a ribosome binding site, and may be codon-optimized for expression in a certain host organism. A method of treating the metabolic disease of PKU by oral administration and ingestion of a GMP is also provided.
1 . A composition for treating phenylketonuria, comprising:
a) a probiotic bacterium; and
b) a gene capable of expressing a functional enzyme for phenylalanine degradation by the probiotic bacterium.
2 . The composition of claim 1 , wherein the probiotic bacterium is selected from the Lactobacilli and Lactococci genera.
3 . The composition of claim 1 , wherein the probiotic bacterium is Lactobacillus reuteri.
4 . The composition of claim 1 , wherein the gene capable of expressing a functional enzyme for phenylalanine degradation is selected from the enzyme family of phenylalanine ammonia-lyases.
5 . The composition of claim 4 , wherein the gene capable of expressing a functional enzyme for phenylalanine degradation is derived from parsley ( Petroselinum crispum ), eubacteria ( Streptomyces maritimus ), cyanobacteria ( Anabaena variabilis, Nostoc punctiforme ) or yeast ( Rhodoturula glutinis ).
6 . The composition of claim 1 , wherein the gene capable of expressing a functional enzyme for phenylalanine degradation is cyanobacterium Anabaena variabilis ATCC 29413.
7 . The composition of claim 1 , wherein the gene capable of expressing a functional enzyme for phenylalanine degradation is codon-optimzed.
8 . The composition of claim 1 , wherein the gene capable of expressing a functional enzyme for phenylalanine degradation includes a promoter.
9 . The composition of claim 8 , wherein the promoter exhibits constitutive expression.
10 . The composition of claim 1 , wherein the gene capable of expressing a functional enzyme for phenylalanine degradation includes a ribosome binding site.
11 . The composition of claim 1 , wherein the gene capable of expressing a functional enzyme for phenylalanine degradation includes a secretion sequence.
12 . The composition of claim 1 , wherein the gene is capable of expressing a functional enzyme for phenylalanine degradation is located in a high retention plasmid within the probiotic.
13 . The composition of claim 1 , wherein the gene capable of expressing functional enzyme for phenylalanine degradation comprises SEQ ID NO:3, or a functional variant thereof.
14 . The composition of claim 1 , wherein the gene capable of expressing functional enzyme for phenylalanine degradation comprises SEQ ID NO:13, or a functional variant thereof.
15 . The composition of claim 1 , wherein the gene capable of expressing functional enzyme for phenylalanine degradation comprises SEQ ID NO:14, or a functional variant thereof.
16 . A method for treating phenylketonuria, comprising the step of:
administering the composition of claim 1 to a patient.
17 . The method of claim 16 , wherein administration is oral.