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A Facile Cell Free Synthesis of Isotopically Labeled (E)-4-Hydroxy-3-methylbut-2-enyl Diphosphate, a Precursor of the Plant Hormone Zeatin

✍ Scribed by Maja Raschke; Monika Fellermeier; Meinhart H. Zenk

Publisher
John Wiley and Sons
Year
2005
Tongue
German
Weight
186 KB
Volume
88
Category
Article
ISSN
0018-019X

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✦ Synopsis

Abstract

(E)‐4‐Hydroxy‐3‐methylbut‐2‐enyl diphosphate (1) is a key intermediate of the deoxyxylulose phosphate pathway of isoprenoid biosynthesis and a precursor of the plant hormone zeatin. The availability of this intermediate with various labeling patterns is pivotal for its use in biosynthetic studies. The number of positions, however, that can be easily labeled by chemical synthesis is limited, and the synthesis by means of recombinant enzymes is laborious and time consuming. We demonstrated that chromoplasts from Capsicum annuum, whose enzyme activity was impaired by freeze‐thawing, accumulate 1. This observation built the basis for the development of a cell‐free system allowing the synthesis of this intermediate with labels in various positions. With 2__C__‐methyl‐D‐erythritol 2,4‐cyclodiphosphate (5) as substrate, yields were in the range of 50%.

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