Synthesis of 4‐methylvaleric acid, a precursor of pogostone, involves a 2‐isobutylmalate synthase related to 2‐isopropylmalate synthase of leucine biosynthesis

Summary We show here that the side chain of pogostone, one of the major components of patchouli oil obtained from Pogostemon cablin and possessing a variety of pharmacological activities, is derived from 4‐methylvaleric acid. We also show that 4‐methylvaleric acid is produced through the one‐carbon α‐ketoacid elongation pathway with the involvement of the key enzyme 2‐isobutylmalate synthase (IBMS), a newly identified enzyme related to isopropylmalate synthase (IPMS) of leucine (Leu) biosynthesis. Site‐directed mutagenesis identified Met 132 in the N‐terminal catalytic region as affecting the substrate specificity of PcIBMS1. Even though PcIBMS1 possesses the C‐terminal domain that in IPMS serves to mediate Leu inhibition, it is insensitive to Leu. The observation of the evolution of IBMS from IPMS, as well as previously reported examples of IPMS‐related genes involved in making glucosinolates in Brassicaceae, acylsugars in Solanaceae, and flavour compounds in apple, indicate that IPMS genes represent an important pool for the independent evolution of genes for specialised metabolism.