The litchi genome has five anthocyanidin reductase (LcANR) and two leucoanthocyanidin reductase (LcLAR) members. The high expression of LcANR1a/2a and LcLAR1/2 is significantly positively correlated with the abundant proanthocyanidins and (-)-epicatechin (EC) in the pericarp, leaf, root, etc. The recombinant LcANR1a/2a converts cyanidin to both EC and (+)-catechin (CT) (EC:CT ≈ 1:1) and converts delphindin to (+)-gallocatechin and (-)-epigallocatechin; the recombinant LcLAR1/2 converts leucocyanidin to CT. The enzymatic kinetics of the four enzymes are presented, with the respective Km of LcLAR1/2 to leucocyanidin, 19 and 34 μM, and the Vmax, 7 and 5 nmol min-1 mg-1, which are rarely reported for other plants. Overexpression of LcANR1a/2a and LcLAR1/2 in Arabidopsis ban mutant recovered EC and CT biosynthesis respectively in the seeds; however, the EC-only recovery by LcANR1a/2a is inconsistent with their in vitro activity, indicating that the ANR/LAR function is dependent on characteristic molecular contexts in plants and correlated to the distinct PA profiles in litchi.
Keywords: anthocyanidin reductase; enzyme kinetics; gene function; leucoanthocyanidin reductase; litchi; proanthocyanidins biosynthesis.