Methods, reagents and cells for biosynthesizing compounds

A method of biosynthesizing a C5 building block selected from the group consisting of glutaric acid, 5-hydroxypentanoic acid, 5-aminopentanoic acid, cadaverine, and 1,5-pentanediol, in a recombinant host, the method comprising:

• (a) (i) enzymatically converting 2-oxo-adipate to 2-amino-adipate using at least one polypeptide having the activity of an alpha-aminotransaminase classified under EC 2.6.1.7 or EC 2.6.1.39, (ii) enzymatically converting 2-oxo-adipate to 5-oxopentanoate using at least one polypeptide having the activity of a 2-oxoacid decarboxylase classified under EC 4.1.1.71, EC 4.1.1.72, EC 4.1.1.43 or EC 4.1.1.74, or (iii) enzymatically converting 2-oxo-adipate to glutaryl-CoA using at least one polypeptide having the activity of a 2-oxoglutarate dehydrogenase complex classified under EC 1.2.4.2, EC 1.8.1.4 and EC 2.3.1.61;
• (b) enzymatically converting the 2-amino-adipate, 5-oxopentanoate, or glutaryl-CoA to the C5 building block, wherein:
• at least one polypeptide having the activity of a thioesterase classified under EC 3.1.2.-, a glutaconate CoA-transferase classified under EC 2.8.3.12, a succinate-CoA ligase classified under EC 6.2.1.5, an aldehyde dehydrogenase classified under EC 1.2.1.-, an acyl-[acp] thioesterase classified under EC 3.1.2.-, a 5-oxopentanoate dehydrogenase classified under EC 1.2.1.-, a 7-oxoheptanoate dehydrogenase classified under EC 1.2.1.-, or a 6-oxohexanoate dehydrogenase classified under EC 1.2.1.63 enzymatically forms a terminal carboxyl group of the C5 building block;
• at least one polypeptide having the activity of an alpha-amino decarboxylase classified under EC 4.1.1.-, a ω-transaminase classified under EC 2.6.1.-, or a deacetylase classified under EC 3.5.1.17 or EC 3.5.1.62 enzymatically forms a terminal amine group of the C5 building block;
• and/or
• at least one polypeptide having the activity of a 6-hydroxyhexanoate dehydrogenase classified under EC 1.1.1.258, a 5-hydroxypentanoate dehydrogenase classified under EC 1.1.1-, a 4-hydroxybutyrate dehydratase classified under EC 1.1.1-, or an alcohol dehydrogenase classified under EC 1.1.1- enzymatically forms a terminal hydroxyl group of the C5 building block.