期刊
PLANTA
卷 210, 期 6, 页码 979-984出版社
SPRINGER VERLAG
DOI: 10.1007/s004250050706
关键词
Arabidopsis; insect cell cultures; jasmonate biosynthesis; 12-oxophytodienoate reductase (overexpression); 12-oxophytodienoate reductase (stereospecificity)
In addition to OPR1 and OPR2, two isoenzymes of 12-oxophytodienoate reductase, a third isoform (OPR3) has recently been identified in Arabidopsis thaliana (L.) Heynh. The expression of the OPR3 gene is induced not only by a variety of stimuli, such as touch. wind, wounding, UV-light and application of detergent, but also by brassinosteroids. The three enzymes were expressed in a functional form in Escherichia coli, and OPR2 was additionally expressed in insect cell cultures and overexpressed in A. thaliana. Substrate conversion was analyzed using a stereospecific assay. The results show that OPR3 effectively converts the natural (9S,13S)-12-oxophytodienoic acid [K-m = 35 mu M, V-max 53.7 nkat (mg protein)(-1)] to the corresponding 3-2(2'(Z)-pentenyl) cyclopentane-1-octanoic acid (OPC-8:0) stereoisomer while OPR1 and OPR2 convert (9S,13S)-12-oxophytodienoic acid with greatly reduced efficiency compared to OPR3. Thus, OPR3 is the isoenzyme relevant for jasmonate biosynthesis.
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