Characterization of a dioxygenase from Trigonella foenum-graecum involved in 4-hydroxyisoleucine biosynthesis.

A study was made of the enzyme in fenugreek implicated in the biosynthesis of 4-hydroxyisoleucine, which is an unusual amino acid known for its insulin stimulating effect. 4-Hydroxyisoleucine was detected by HPLC following isoleucine incubation with a cell-free extract from etiolated 6-day-old fenugreek seedlings in the presence of various cofactors. The reaction showed that 4-hydroxyisoleucine formation is dependent on the presence of Fe2+, 2-oxoglutarate, ascorbate and oxygen. This suggests that a 2-oxoacid dependent dioxygenase plays a key role in this biosynthetic pathway.