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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
Author(s) -
Raschke Maja,
Fellermeier Monika,
Zenk Meinhart H.
Publication year - 2005
Publication title -
helvetica chimica acta
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.74
H-Index - 82
eISSN - 1522-2675
pISSN - 0018-019X
DOI - 10.1002/hlca.200590116
Subject(s) - chemistry , zeatin , biosynthesis , enzyme , biochemistry , substrate (aquarium) , stereochemistry , chemical synthesis , auxin , cytokinin , in vitro , oceanography , gene , geology
( 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%.