RESUMEN
Photosynthetic organisms such as plants, algae and some cyanobacteria synthesize tocochromanols, a group of compounds that encompasses tocopherols and tocotrienols and that exhibits vitamin E activity in animals. While most vitamin E biosynthetic genes have been identified in plant genomes, regulatory genes controlling tocopherol accumulation are currently unknown. We isolated by forward genetics Arabidopsis enhanced vitamin E (eve) mutants that overaccumulate the classic tocopherols and plastochromanol-8, and a tocochromanol unknown in this species. We mapped eve1 and eve4, and identified the unknown Arabidopsis tocochromanol by using a combination of analytical tools. In addition, we determined its biosynthetic pathway with a series of tocochromanol biosynthetic mutants and transgenic lines. eve1 and eve4 are two seed lipid mutants affecting the WRINKLED1 (WRI1) and ACYL-COA:DIACYLGLYCEROL ACYLTRANSFERASE1 (DGAT1) genes, respectively. The unknown tocochromanol is 11'-12' γ-tocomonoenol, whose biosynthesis is VITAMIN E 1 (VTE1) - and VTE2-dependent and is initiated by the condensation of homogentisate (HGA) and tetrahydrogeranylgeranyl pyrophosphate. This study identifies the first two regulatory genes, WRI1 and DGAT1, that control the synthesis of all tocochromanol forms in seeds, and shows the existence of a metabolic trade-off between lipid and tocochromanol metabolisms. Moreover, it shows that Arabidopsis possesses a tocomonoenol biosynthetic pathway that competes with tocopherol synthesis.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimología , Diacilglicerol O-Acetiltransferasa/metabolismo , Diglicéridos/metabolismo , Factores de Transcripción/metabolismo , Vitamina E/metabolismo , Acilcoenzima A/metabolismo , Transferasas Alquil y Aril/genética , Transferasas Alquil y Aril/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Vías Biosintéticas , Cromanos/metabolismo , Diacilglicerol O-Acetiltransferasa/genética , Transferasas Intramoleculares/genética , Transferasas Intramoleculares/metabolismo , Metabolismo de los Lípidos , Semillas/enzimología , Semillas/genética , Tocoferoles/metabolismo , Tocotrienoles/metabolismo , Factores de Transcripción/genética , Vitamina E/análogos & derivadosRESUMEN
Tocochromanols are organic compounds mostly produced by photosynthetic organisms that exhibit vitamin E activity in animals. They result from the condensation of homogentisate with four different polyprenyl side chains derived all from geranylgeranyl pyrophosphate. The core tocochromanol biosynthesis has been investigated in several photosynthetic organisms and is now well-characterized. In contrast, our current knowledge of the biosynthesis and transport of tocochromanol biosynthetic precursors is much more limited. While tocochromanol synthesis occurs in plastids, converging genetic data in Arabidopsis and soybean demonstrate that the synthesis of the polar precursor homogentisate is located in the cytoplasm. These data implies that tocochromanol synthesis involves several plastidic membrane transporter(s) that remain to be identified. In addition, the metabolic origin of the lipophilic isoprenoid precursor is not fully elucidated. While some genetic data exclusively attribute the synthesis of the prenyl component of tocochromanols to the plastidic methyl erythritol phosphate pathway, multiple lines of evidence provided by feeding experiments and metabolic engineering studies indicate that it might partially originate from the cytoplasmic mevalonate pathway. Although this question is still open, these data demonstrate the existence of membrane transporter(s) capable of importing cytosolic polyprenyl pyrophosphate such as farnesyl pyrophosphate into plastids. Since the availability of both homogentisate and polyprenyl pyrophosphates are currently accepted as major mechanisms controlling the type and amount of tocochromanols produced in plant tissues, we summarized our current knowledge and research gaps concerning the biosynthesis, metabolic origins and transport of tocochromanol biosynthetic precursors in plant cells.
RESUMEN
Vitamin E refers to four tocopherols and four tocotrienols that are exclusively synthesized by photosynthetic organisms. While α-tocopherol is the most potent vitamin E compound, it is not the main form consumed since the composition of most major crops is dominated by γ-tocopherol. Nutritional studies show that populations of developed countries do not consume enough vitamin E and that a large proportion of individuals exhibit plasma α-tocopherol deficiency. Following the identification of vitamin E biosynthetic genes, several strategies including metabolic engineering, classic breeding and mutation breeding, have been undertaken to improve the vitamin E content of crops. In addition to providing crops in which vitamin E content is enhanced, these studies are revealing the bottlenecks limiting its biosynthesis.