Host cells and methods for producing toluene biochemically

A genetically modified host cell comprising a first nucleic acid encoding a first polypeptide operatively linked a first promoter, wherein the first polypeptide comprises an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2 and a conserved qlycyl radical motif comprising R at position 812, V at position 813, G at position 815, F at position 816, L at position 823, Q at position 828, 1 at position 831, and R at position 834, and a conserved C at position 482, of the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2, and having an enzymatic activity to decarboxylate a phenylacetic acid into a toluene and a carbon dioxide;

• and the first nucleic acid, or a second nucleic acid, encoding a second polypeptide operatively linked to the first promoter or a second promoter, wherein the second polypeptide comprises an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence of SEQ ID NO:3 or SEQ ID NO:4 and conserved amino acid residues:
• C at position 33, C at position 37, and C at position 40 of the amino acid sequence of SEQ ID NO:3, or C at position 39, C at position 43, and C at position 46 of the amino acid sequence of SEQ ID NO:4, and having an enzymatic activity to cleave a S-adenosylmethionine (SAM) to form a methionine and a 5′-deoxyadenosyl radical;
• wherein the genetically modified host cell is capable of expressing the first and the second polypeptide and the first polypeptide or the second polypeptide is heterologous to (i) the genetically modified host cell, or (ii) the first promoter or the second promoter.