植物维生素E生物强化研究进展

维生素E是一种只能在光合组织中合成的脂溶性小分子有机化合物,是人体和动物营养不可缺少的重要维生素。由于植物中维生素E含量较低,人类大多处于慢性缺乏维生素E--“隐性饥饿”的状态,而动物饲料中则需要添加外源合成的维生素E以满足其营养需求。因此,提高植物中维生素E的含量是改善维生素E缺乏的重要途径之一。从维生素E的合成途径入手,详细地综述了维生素E合成关键酶基因的表达变化以及前体物质的含量变化对维生素E合成的影响,发现三烯生育酚和α-生育酚的生物强化效果较好,而生育酚总量提高受限;进而从遗传的角度探讨了维生素E合成受限的原因以及遗传上可能影响维生素E合成的其他代谢途径;最后结合可能影响维生素E合成的调控因子以及其前体物质的转运等方面为今后维生素E的生物强化提出了新的思路。     

生育三烯酚的生理功能及合成代谢调控

维生素E是一种重要的脂溶性抗氧化剂。天然维生素E包括α-、β-、γ-和δ-生育酚及α-、β-、γ-和δ-生育三烯酚等8种组分。生育三烯酚具有独特的降低胆固醇、抗癌和神经保护功能,这些功能是生育酚所不具备的。生育三烯酚主要由谷物等单子叶植物合成。虽然生育三烯酚成分占天然维生素E的一半,但有关生育三烯酚的研究仅占维生素E全部文献的1%。可喜的是,近年来这一领域的研究取得了较大的进展。我们就生育三烯酚在人体内的生理功能、来源、生物合成及代谢调控等方面的进展进行概述。     

天然维生素E的研究进展

维生素E是一类脂溶性、具抗氧化功能的维生素,按其来源可分为天然维生素E和人工合成维生素E.对天然维生素E的功能、生物合成途径以及相关酶基因的研究方面进行了综述.其中,维生素E合成相关酶基因已经克隆及定位,尤其VTE5的发现,为生育酚合成研究开辟了一条新途径.人们已经开始利用基因工程技术研究提高植物天然维生素E产量的方法.     

植物维生素E基因工程研究进展

维生素E(vitamin E)是一种脂溶性维生素,是生物体内最主要的抗氧化剂之一。本文综述了植物维生素E的合成途径及所涉及的相关基因,针对提高植物中维生素E含量的基因工程改造,提出三条途径,即提高代谢底物水平、提高植物生育酚总量和提高α 生育酚比例,分析了维生素E基因工程改造过程中需要考虑的因素,最后展望了未来作物维生素E的品质改良的研究方向。     

尿苷对线粒体功能的影响

尿苷是生物体内必需的营养物之一,需将尿苷浓度维持在一定浓度水平才能维持胞内正常的生长代谢。近年来,有研究发现尿苷可通过多种机制缓解生物体炎症反应,并能参与胞内糖酵解等代谢途径,且可调节胞内糖基化、乙酰化等蛋白修饰作用。此外,其还能通过减轻胞内氧化应激、促进高能化合物合成等方式保护细胞免受缺氧性损伤。研究表明,这些保护作用与尿苷对线粒体功能的调节作用息息相关。因此,本文主要综述了尿苷及其代谢反应(物)对线粒体功能的研究进展。     

三烯生育酚生物学功能及合成调控研究进展

介绍了三烯生育酚的生物合成途径,重点综述了三烯生育酚在神经保护、抗癌、降低胆固醇以及抗氧化等方面的优越生物学功能,以及利用关键酶的高效表达和前体物质水平的提高等植物代谢工程手段提高植物体内三烯生育酚生物合成水平的研究进展。     

植物维生素E基因工程研究进展

维生素E(vitamin E, VE)是一类由光合生物合成的、人类饮食中必不可少的两种抗氧化物质,分为生育酚和生育三烯酚两大类。除了生育酚类物质所具有的抗氧化作用外,生育三烯酚还有很强的降胆固醇、预防糖尿病、促进骨吸收、抗癌和神经保护的作用,因此,VE被广泛应用于医药、食品、化妆品等行业中。本文主要综述了植物维生素E生物合成相关酶的研究进展以及利用基因工程手段提高植物维生素E活性的新策略。其中,多基因共转化、多基因操纵子及质体转化等方法为提高植物维生素E活性提供了新的思路。     

植物中维生素E的生物学功能研究进展

维生素E又称生育酚,是人类和动物营养所必需的一类脂溶性的维生素。维生素E只能在植物和光合细菌中合成,人类和动物只能通过摄入植物性食物或药物进行补充。与人类和动物相比,植物中维生素E的生物学功能研究进展相对缓慢。近些年得益于转基因植株或者植物突变体材料的获得,维生素E在植物体内的功能研究得到了迅速发展。综述了维生素E在种子萌发、植株生长发育、衰老过程、糖类运输、信号转导以及对逆境的响应等方面的生理功能,并对未来的研究提出了展望,以期为维生素E的基础和应用研究提供参考。     

微量营养素与抗氧化防御作用

现代医学证明,机体和氧进行正常交换时产生的自由基,能损伤蛋白质、核酸等生物大分子,改变细胞结构与功能,从而影响人体健康、引发疾病、甚至危及生命。微量营养素中的维生素E、维生素C、β-胡萝卜素等维生素和某些抗氧化酶依赖的硒、锌、铜等元素,是生物体内抗氧化防御系统的组成成分,可直接或间接地清除活性氧或阻断连锁氧化反应,保护细胞及生物大分子免受氧化损伤。因此,它们在抗氧化防御系统中具有重要的作用,合理摄入这些微量营养素,对预防动脉硬化、抗癌、改善免疫功能及延缓衰老等有着特殊的意义。     

利用基因工程提高植物维生素E营养品质的策略

维生素E是一种对植物、动物和人类都具有重要作用的脂溶性维生素。而植物则是人类维生素E的主要来源。对维生素E的结构和合成途径进行了简单的介绍,并重点综述了利用基因工程提高植物维生素E营养品质的策略。这些策略主要包括导入编码影响维生素E总量相关酶的基因来提高维生素E的总量;导入编码影响维生素E组成相关酶的基因,提高α-生育酚在总生育酚中所占的比例,从而提高维生素E的活性。     

Vitamin E biosynthesis: functional characterization of the monocot homogentisate geranylgeranyl transferase

The biosynthesis of the tocotrienol and tocopherol forms of vitamin E is initiated by prenylation of homogentisate. Geranylgeranyl diphosphate (GGDP) is the prenyl donor for tocotrienol synthesis, whereas phytyl diphosphate (PDP) is the prenyl donor for tocopherol synthesis. We have previously shown that tocotrienol synthesis is initiated in monocot seeds by homogentisate geranylgeranyl transferase (HGGT). This enzyme is related to homogentisate phytyltransferase (HPT), which catalyzes the prenylation step in tocopherol synthesis. Here we show that monocot HGGT is localized in the plastid and expressed primarily in seed endosperm. Despite the close structural relationship of monocot HGGT and HPT, these enzymes were found to have distinct substrate specificities. Barley (Hordeum vulgare cv. Morex) HGGT expressed in insect cells was six times more active with GGDP than with PDP, whereas the Arabidopsis HPT was nine times more active with PDP than with GGDP. However, only small differences were detected in the apparent Km values of barley HGGT for GGDP and PDP. Consistent with its in vitro substrate properties, barley HGGT generated a mixture of tocotrienols and tocopherols when expressed in the vitamin E-null vte2-1 mutant lacking a functional HPT. Relative levels of tocotrienols and tocopherols produced in vte2-1 differed between organs and growth stages, reflective of the composition of plastidic pools of GGDP and PDP. In addition, HGGT was able to functionally substitute for HPT to rescue vte2-1-associated phenotypes, including reduced seed viability and increased fatty acid oxidation of seed lipids. Overall, we show that monocot HGGT is biochemically distinct from HPT, but can replace HPT in important vitamin E-related physiological processes.     

Genetic and biochemical basis for alternative routes of tocotrienol biosynthesis for enhanced vitamin E antioxidant production

Vitamin E tocotrienol synthesis in monocots requires homogentisate geranylgeranyl transferase (HGGT), which catalyzes the condensation of homogentisate and the unsaturated C20 isoprenoid geranylgeranyl diphosphate (GGDP). By contrast, vitamin E tocopherol synthesis is mediated by homogentisate phytyltransferase (HPT), which condenses homogentisate and the saturated C20 isoprenoid phytyl diphosphate (PDP). An HGGT‐independent pathway for tocotrienol synthesis has also been shown to occur by de‐regulation of homogentisate synthesis. In this paper, the basis for this pathway and its impact on vitamin E production when combined with HGGT are explored. An Arabidopsis line was initially developed that accumulates tocotrienols and homogentisate by co‐expression of Arabidopsis hydroxyphenylpyruvate dioxygenase (HPPD) and Escherichia coli bi‐functional chorismate mutase/prephenate dehydrogenase (TyrA). When crossed into the vte2–1 HPT null mutant, tocotrienol production was lost, indicating that HPT catalyzes tocotrienol synthesis in HPPD/TyrA‐expressing plants by atypical use of GGDP as a substrate. Consistent with this, recombinant Arabidopsis HPT preferentially catalyzed in vitro production of the tocotrienol precursor geranylgeranyl benzoquinol only when presented with high molar ratios of GGDP:PDP. In addition, tocotrienol levels were highest in early growth stages in HPPD/TyrA lines, but decreased strongly relative to tocopherols during later growth stages when PDP is known to accumulate. Collectively, these results indicate that HPPD/TyrA‐induced tocotrienol production requires HPT and occurs upon enrichment of GGDP relative to PDP in prenyl diphosphate pools. Finally, combined expression of HPPD/TyrA and HGGT in Arabidopsis leaves and seeds resulted in large additive increases in vitamin E production, indicating that homogentisate concentrations limit HGGT‐catalyzed tocotrienol synthesis.     

Anti-angiogenic activity of tocotrienol

The anti-angiogenic property of vitamin E compounds, with particular emphasis on tocotrienol, has been investigated in vitro. Tocotrienol, but not tocopherol, inhibited both the proliferation and tube formation of bovine aortic endothelial cells, with delta-tocotrienol appearing the highest activity. Also, delta-tocotrienol reduced the vascular endothelial growth factor-stimulated tube formation by human umbilical vein endothelial cells. Our findings suggest that tocotrienol has potential use as a therapeutic dietary supplement for minimizing tumor angiogenesis.     

Anti-inflammatory properties of alpha- and gamma-tocopherol

Natural vitamin E consists of four different tocopherol and four different tocotrienol homologues (alpha,beta, gamma, delta) that all have antioxidant activity. However, recent data indicate that the different vitamin E homologues also have biological activity unrelated to their antioxidant activity. In this review, we discuss the anti-inflammatory properties of the two major forms of vitamin E, alpha-tocopherol (alphaT) and gamma-tocopherol (gammaT), and discuss the potential molecular mechanisms involved in these effects. While both tocopherols exhibit anti-inflammatory activity in vitro and in vivo, supplementation with mixed (gammaT-enriched) tocopherols seems to be more potent than supplementation with alphaT alone. This may explain the mostly negative outcomes of the recent large-scale interventional chronic disease prevention trials with alphaT only and thus warrants further investigation.     

Anti-angiogenic Potential of Tocotrienolin vitro

Modulation of angiogenesis is now a recognized strategy for the prevention of various angiogenesis-mediated disorders. We investigated, using well-characterized in vitro systems, the anti-angiogenic property of vitamin E compounds, with particular emphasis on tocotrienol, a natural analog of tocopherol. Tocotrienol, but not tocopherol, inhibited the proliferation of bovine aortic endothelial cells in dose dependent manner at half-maximal concentrations in the low micromolar range. Tocotrienol also significantly inhibited the formation of networks of elongated endothelial cells within 3D collagen gels. From these results, we suggest that tocotrienol is a potential candidate for the development of useful therapeutic agents or preventive food factors for tumor angiogenesis.     

Does lack of tocopherols and tocotrienols put women at increased risk of breast cancer?

Breast cancer is the leading site of new cancers in women and the second leading cause (after lung cancer) of cancer mortality in women. Observational studies that have collected data for dietary exposure to alpha-tocopherol with or without the other related tocopherols and tocotrienols have suggested that vitamin E from dietary sources may provide women with modest protection from breast cancer. However, there is no evidence that vitamin E supplements confer any protection whatever against breast cancer. Observational studies that have assessed exposure to vitamin E by plasma or adipose tissue concentrations of alpha-tocopherol have failed to provide consistent support for the idea that alpha-tocopherol provides any protection against breast cancer. In addition, evidence from studies in experimental animals suggest that alpha-tocopherol supplementation alone has little effect on mammary tumors. In contrast, studies in breast cancer cells indicate that alpha- gamma-, and delta-tocotrienol, and to a lesser extent delta-tocopherol, have potent antiproliferative and proapoptotic effects that would be expected to reduce risk of breast cancer. Many vegetable sources of alpha-tocopherol also contain other tocopherols or tocotrienols. Thus, it seems plausible that the modest protection from breast cancer associated with dietary vitamin E may be due to the effects of the other tocopherols and the tocotrienols in the diet. Additional studies will be required to determine whether this may be the case, and to identify the most active tocopherol/tocotrienol.     

Metabolic redesign of vitamin E biosynthesis in plants for tocotrienol production and increased antioxidant content

Tocotrienols are the primary form of vitamin E in seeds of most monocot plants, including cereals such as rice and wheat. As potent antioxidants, tocotrienols contribute to the nutritive value of cereal grains in human and livestock diets. cDNAs encoding homogentisic acid geranylgeranyl transferase (HGGT), which catalyzes the committed step of tocotrienol biosynthesis, were isolated from barley, wheat and rice seeds. Transgenic expression of the barley HGGT in Arabidopsis thaliana leaves resulted in accumulation of tocotrienols, which were absent from leaves of nontransformed plants, and a 10- to 15-fold increase in total vitamin E antioxidants (tocotrienols plus tocopherols). Overexpression of the barley HGGT in corn seeds resulted in an increase in tocotrienol and tocopherol content of as much as six-fold. These results provide insight into the genetic basis for tocotrienol biosynthesis in plants and demonstrate the ability to enhance the antioxidant content of crops by introduction of an enzyme that redirects metabolic flux.     

Transgenic rice grains expressing a heterologous ρ-hydroxyphenylpyruvate dioxygenase shift tocopherol synthesis from the γ to the α isoform without increasing absolute tocopherol levels

We generated transgenic rice plants overexpressing Arabidopsis thaliana ρ-hydroxyphenylpyruvate dioxygenase (HPPD), which catalyzes the first committed step in vitamin E biosynthesis. Transgenic grains accumulated marginally higher levels of total tocochromanols than controls, reflecting a small increase in absolute tocotrienol synthesis (but no change in the relative abundance of the α and γ isoforms). In contrast, there was no change in the absolute tocopherol level, but a significant shift from the γ to the α isoform. These data confirm HPPD is not rate limiting, and that increasing flux through the early pathway reveals downstream bottlenecks that act as metabolic tipping points.