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The metabolite chemotype of N icotiana benthamiana transiently expressing artemisinin biosynthetic pathway genes is a function of CYP 71 AV 1 type and relative gene dosage
Author(s) -
Ting HiengMing,
Wang Bo,
Rydén AnnaMargareta,
Woittiez Lotte,
Herpen Teun,
Verstappen Francel W. A.,
RuyterSpira Carolien,
Beekwilder Jules,
Bouwmeester Harro J.,
Krol Alexander
Publication year - 2013
Publication title -
new phytologist
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.742
H-Index - 244
eISSN - 1469-8137
pISSN - 0028-646X
DOI - 10.1111/nph.12274
Subject(s) - chemotype , artemisinin , nicotiana benthamiana , biosynthesis , biology , biochemistry , secondary metabolite , enzyme , gene , artemisia annua , chemistry , plasmodium falciparum , botany , malaria , essential oil , immunology
SummaryA rtemisia annua , which produces the anti‐malaria compound artemisinin, occurs as high‐artemisinin production ( HAP ) and low‐artemisinin production ( LAP ) chemotypes. Understanding the basis of the difference between these chemotypes would assist breeding and optimising artemisinin biosynthesis. Here we present a systematic comparison of artemisinin biosynthesis genes that may be involved in determining the chemotype ( CYP 71 AV 1 , DBR 2 and ALDH 1 ). These genes were isolated from the two chemotypes and characterized using transient expression in planta . The enzyme activity of DBR 2 and ALDH 1 from the two chemotypes did not differ, but structural differences in CYP 71 AV 1 from LAP and HAP chemotypes ( AMOLAP and AMOHAP , respectively) resulted in altered enzyme activity. AMOLAP displays a seven amino acids N ‐terminal extension compared with AMOHAP . The GFP fusion of both proteins show equal localization to the ER but AMOHAP may have reduced stability. Upon transient expression in N icotiana benthamiana , AMOLAP displayed a higher enzyme activity than AMOHAP . However, expression in combination with the other pathway genes also resulted in a qualitatively different product profile (‘chemotype’); that is, in a shift in the ratio between the unsaturated and saturated (dihydro) branch of the pathway.