Polypeptides and biosynthetic pathways.

摘要

Procedure producing monatin, comprising converting tryptophan and / or indole-3láctico acid to monatin, wherein the method comprises: (a) contacting tryptophan or indole-3láctico acid with a first polypeptide, wherein, when tryptophan is contacted with the first polypeptide, the first polypeptide is chosen from tryptophan aminotransferase (EC 2.6.1.27), tryptophan dehydrogenase (EC 1.4.1.19), tyrosine aminotransferase (aromatic) (EC 2.6.1.5 ), tryptophan-phenylpyruvate transaminase (EC 2.6.1.28), multiple substrate aminotransferase (EC 2.6.1.-), aspartate aminotransferase (EC 2.6.1.1), tryptophan oxidase, L-amino acid oxidase (EC 1.4.3.2), D amino acid dehydrogenase (EC 1.4.99.1), D-amino acid oxidase (EC 1.4.3.3), tryptophan aminotransferase D-, D-alanine aminotransferase (EC 2.6.1.21), glutamate dehydrogenase (EC 1.4.1.2-4), phenylalanine dehydrogenase ( EC 1.4.1.20), or a combination thereof, in the q ue, when the three-lactic-indole is contacted with the first polypeptide, the first polypeptide is selected from indol-lactate dehydrogenase (EC 1.1.1.110), R-4-hydroxyphenyl-lactate dehydrogenase (EC 1.1.1.222) , 3- (4) -hydroxyphenylpyruvate reductase (EC 1.1.1.237), lactate dehydrogenase (EC 1.1.1.27, 1.1.1.28, 1.1.2.3), (3-imidazol-5-yl) lactate dehydrogenase (EC 1.1.1.111) , and lactate oxidase (EC 1.1.3.-), or a combination thereof, wherein the first polypeptide converts tryptophan or indole-3láctico acid to indole-3-pyruvate; (B) contacting the indole-3-pyruvate with a second polypeptide and C3 carbon source, wherein the second polypeptide is an EC 4.1.2 or 4.1.3 enzyme EC., Wherein the second polypeptide converts indole-3-pyruvate and the C3 carbon source to 2-hydroxy 2- (indol-3-ylmethyl) -4-keto glutaric acid; and (c) contacting the 2-hydroxy 2- (indol-3-ylmethyl) -4-keto glutaric acid with a third polypeptide, wherein the third polypeptide is chosen from tyrosine aminotransferase (aromatic) (EC 2.6.1.5) , tryptophan dehydrogenase (EC 1.4.1.19), tryptophan-phenylpyruvate transaminase (EC 2.6.1.28), tryptophan aminotransferase (EC 2.6.1.27), aspartate aminotransferase (EC 2.6.1.1), D-alanine aminotransferase (EC 2.6.1.21), D-tryptophan aminotransferase, glutamate dehydrogenase (EC 1.4.1.2-4), phenylalanine dehydrogenase (EC 1.4.1.20), multiple substrate aminotransferase (EC 2.6.1.-), D-amino acid dehydrogenase (EC 1.4.99.1), a lysine epsilon aminotransferase, or a combination thereof, wherein the third polypeptide converts the 2-hydroxy 2- (indol-3-ylmethyl) -4-keto glutaric acid to monatin.