5-Dehydroshikimate reductase in the tea plant (Camellia sinensis L.). Properties and distribution

Abstract

A method for the extraction of 5-dehydroshiMmate reductase (EC 1.1.1.25) from tea plant tissues in an active soluble state has been developed. It is dependent on the use, in the extraction medium, of an insoluble polyphenol adsorbent (Polyclar AT), which prevents the polyphenols present from precipitating all the proteins. The enzyme has the following properties: pH optima at pH10-1 in glycine-sodium hydroxide buffer and at pH 7.7 in tris-hydrochloric acid buffer; Km (NADP) 32 [mu][image] and Km (shikimate) 0.43m[image]; and NADP-specificity. It was completely inhibited by 0.33m[image] -p-chloromer curibenzoate and this inhibition was completely reversed by 10m[image]-dysteine. Iodoacetate and arsenite inhibited the enzyme to a smaller extent. The specific activity of the enzyme was higher in the parts of the actively growing shoot tips (third leaf > stem >second leaf >first leaf >bud) than in the mature leaves. However, the mature leaves had the greatest total activity. The importance of these findings with respect to flavanol biosynthesis in tea plants is discussed.