Kinetic mechanism of NADH‐enoyl‐ACP reductase from Brassica napus

Abstract
Enoyl‐ACP reductase, a component of fatty acid synthase, is a target for anti‐microbial agents and herbicides. Here we demonstrate the kinetic mechanism to be a compulsory‐order ternary complex with NADH binding before the acyl substrate. Matrix‐assisted laser desorption ionisation mass spectrometry analysis of enzymatically and synthesised crotonyl‐ACP substrate showed the former to contain a single acyl group, whereas the latter contained up to four additional crotonylations. The use of authentic crotonyl‐ACP will be important in future kinetic and crystallographic studies.

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