RESUMEN
The antioxidant responses of coffee (Coffea arabica L.) cell suspension cultures to nickel (Ni) were investigated. Ni was very rapidly accumulated in the cells and the accumulation could be directly correlated with the increase of NiCl(2) concentration in the medium. At 0.05 mM NiCl(2) growth was stimulated, but at 0.5 mM NiCl(2), the growth rate was reduced. An indication of alterations in the presence of reactive oxygen species was detected by an increase in lipid peroxidation at 0.5 mM NiCl(2). Catalase (CAT; EC 1.11.1.6), glutathione reductase (GR; EC 1.6.4.2), ascorbate peroxidase (APX; EC 1.11.1.11), guaiacol peroxidase (GOPX; EC 1.11.1.7) and superoxide dismutase (SOD; EC 1.15.1.1) activities were increased, particularly at earlier NiCl(2) exposure times and the activities were higher at 0.5 mM NiCl(2) for most of exposure times tested. Non-denaturing PAGE revealed one CAT isoenzyme, nine SOD isoenzymes and four GR isoenzymes. The SOD isoenzymes were differentially affected by NiCl(2) treatment and one GR isoenzyme was increased by NiCl(2). NiCl(2) at 0.05 mM did not induce lipid peroxidation and the main response appeared to be via the induction of SOD, CAT, GOPX and APX activities for the removal of the reactive oxygen species and through the induction of GR to ensure the availability of reduced glutathione.
Asunto(s)
Antioxidantes/metabolismo , Coffea/efectos de los fármacos , Níquel/farmacología , Células Cultivadas , Coffea/citología , Coffea/metabolismo , Activación Enzimática , Glutatión/metabolismo , Glutatión Reductasa/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Peroxidasas/metabolismo , Proteínas de Plantas/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Superóxido Dismutasa/metabolismoRESUMEN
Aspartate is the common precursor of the essential amino acids lysine, threonine, methionine and isoleucine in higher plants. In addition, aspartate may also be converted to asparagine, in a potentially competing reaction. The latest information on the properties of the enzymes involved in the pathways and the genes that encode them is described. An understanding of the overall regulatory control of the flux through the pathways is undisputedly of great interest, since the nutritive value of all cereal and legume crops is reduced due to low concentrations of at least one of the aspartate-derived amino acids. We have reviewed the recent literature and discussed in this paper possible methods by which the concentrations of the limiting amino acids may be increased in the seeds.
Asunto(s)
Ácido Aspártico/metabolismo , Plantas/metabolismo , Isoleucina/biosíntesis , Lisina/metabolismo , Plantas/enzimologíaRESUMEN
The essential amino acids lysine and threonine are synthesized in higher plants via a pathway starting with aspartate that also leads to the formation of methionine and isoleucine. Lysine is one of most limiting amino acids in plants consumed by humans and livestock. Recent genetic, molecular, and biochemical evidence suggests that lysine synthesis and catabolism are regulated by complex mechanisms. Early kinetic studies utilizing mutants and transgenic plants that over-accumulate lysine have indicated that the major step for the regulation of lysine biosynthesis is at the enzyme dihydrodipicolinate synthase. Despite this tight regulation, recent strong evidence indicates that lysine catabolism is also subject to control, particularly in cereal seeds. The challenge of producing crops with a high-lysine concentration in the seeds appeared to be in sight a few years ago. However, apart from the quality protein maize lines currently commercially available, the release of high-lysine crops has not yet occurred. We are left with the question, is the production of high-lysine crops still a challenge?
Asunto(s)
Productos Agrícolas/metabolismo , Lisina/biosíntesis , Treonina/biosíntesis , Grano Comestible/metabolismo , Humanos , Plantas Modificadas Genéticamente/metabolismoRESUMEN
The essential amino acids lysine and threonine are synthesized in higher plants via a pathway starting with aspartate that also leads to the formation of methionine and isoleucine. Lysine is one of most limiting amino acids in plants consumed by humans and livestock. Recent genetic, molecular, and biochemical evidence suggests that lysine synthesis and catabolism are regulated by complex mechanisms. Early kinetic studies utilizing mutants and transgenic plants that over-accumulate lysine have indicated that the major step for the regulation of lysine biosynthesis is at the enzyme dihydrodipicolinate synthase. Despite this tight regulation, recent strong evidence indicates that lysine catabolism is also subject to control, particularly in cereal seeds. The challenge of producing crops with a high-lysine concentration in the seeds appeared to be in sight a few years ago. However, apart from the quality protein maize lines currently commercially available, the release of high-lysine crops has not yet occurred. We are left with the question, is the production of high-lysine crops still a challenge?.
Asunto(s)
Humanos , Productos Agrícolas/metabolismo , Lisina/biosíntesis , Treonina/biosíntesis , Grano Comestible/metabolismo , Plantas Modificadas Genéticamente/metabolismoRESUMEN
Lysine is catabolyzed by the bifunctional enzyme lysine 2-oxoglutarate reductase-saccharopine dehydrogenase (LOR-SDH) in both animals and plants. LOR condenses lysine and 2-oxoglutarate into saccharopine, using NADPH as cofactor and SDH converts saccharopine into alpha-aminoadipate delta-semialdehyde and glutamic acid, using NAD as cofactor. The distribution pattern of LOR and SDH among different tissues of Phaseolus vulgaris was determined. The hypocotyl contained the highest specific activity, whereas in seeds the activities of LOR and SDH were below the limit of detection. Precipitation of hypocotyl proteins with increasing concentrations of PEG 8000 revealed one broad peak of SDH activity, indicating that two isoforms may be present, a bifunctional LOR-SDH and possibly a monofunctional SDH. During the purification of the hypocotyl enzyme, the LOR activity proved to be very unstable, following ion-exchange chromatography. Depending on the purification procedure, the protein eluted as a monomer of 91-94 kDa containing only SDH activity, or as a dimer of 190 kDa with both, LOR and SDH activities, eluting together.
Asunto(s)
Phaseolus/enzimología , Sacaropina Deshidrogenasas/aislamiento & purificación , Peso Molecular , Sacaropina Deshidrogenasas/químicaRESUMEN
To investigate the antioxidant responses of radish (Raphanus sativus L.) to cadmium (Cd) treatment, seedlings of a tolerant variety were grown in increasing concentrations of CdCl(2), ranging from 0.25-1 mM, for up to 72 h in a hydroponic system. Analysis of Cd uptake indicated that most of the Cd accumulated in the roots, but some was also translocated and accumulated in the leaves, especially at the higher concentrations of Cd used in the experiments. Roots and leaves were analysed for catalase, glutathione reductase and superoxide dismutase activities. Catalase and glutathione reductase activities increased considerably in the roots and leaves after 24 h exposure to the metal, indicating a direct correlation with Cd accumulation. The analysis of native PAGE enzyme activity staining, revealed several superoxide dismutase isoenzymes in leaves, with the two predominant isoenzymes exhibiting increases in activity in response to Cd treatment. The results suggest that in radish, the activity of antioxidant enzymes responds to Cd treatment. The main response may be via the activation of the ascorbate-glutathione cycle for the removal of hydrogen peroxide, or to ensure the availability of glutathione for the synthesis of Cd-binding proteins.
Asunto(s)
Brassica/efectos de los fármacos , Cadmio/farmacología , Catalasa/metabolismo , Glutatión Reductasa/metabolismo , Superóxido Dismutasa/metabolismo , Brassica/enzimología , Hojas de la Planta/enzimología , Raíces de Plantas/enzimologíaRESUMEN
The essential amino acid lysine is synthesised in higher plants via a pathway starting with aspartate, that also leads to the formation of threonine, methionine and isoleucine. Enzyme kinetic studies and the analysis of mutants and transgenic plants that overaccumulate lysine, have indicated that the major site of the regulation of lysine synthesis is at the enzyme dihydrodipicolinate synthase. Despite this tight regulation, there is strong evidence that lysine is also subject to catabolism in plants, specifically in the seed. The two enzymes involved in lysine breakdown, lysine 2-oxoglutarate reductase (also known as lysine a-ketoglutarate reductase) and saccharopine dehydrogenase exist as a single bifunctional protein, with the former activity being regulated by lysine availability, calcium and phosphorylation/ dephosphorylation.
Asunto(s)
Lisina/metabolismo , Plantas/metabolismo , Mutación , Plantas/genética , Plantas Modificadas Genéticamente/metabolismoRESUMEN
The essential amino acids lysine, threonine, methionine and isoleucine are synthesised in higher plants via a common pathway starting with aspartate. The regulation of the pathway is discussed in detail, and the properties of the key enzymes described. Recent data obtained from studies of regulation at the gene level and information derived from mutant and transgenic plants are also discussed. The herbicide target enzyme acetohydroxyacid synthase involved in the synthesis of the branched chain amino acids is reviewed.