Polyprenyl pyrophosphate-p-hydroxybenzoate polyprenyltransferase activity in mitochondria of broad-bean seeds and yeast.
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ABSTRACT:
SUBMITTER: Thomas G
PROVIDER: S-EPMC1177878 | biostudies-other | 1973 Jul
REPOSITORIES: biostudies-other
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Project description:Ubiquinone is an essential component of the electron transfer system in both prokaryotes and eukaryotes and is synthesized from chorismate and polyprenyl diphosphate by eight steps. p-Hydroxybenzoate (PHB) polyprenyl diphosphate transferase catalyzes the condensation of PHB and polyprenyl diphosphate in ubiquinone biosynthesis. We isolated the gene (designated ppt1) encoding PHB polyprenyl diphosphate transferase from Schizosaccharomyces pombe and constructed a strain with a disrupted ppt1 gene. This strain could not grow on minimal medium supplemented with glucose. Expression of COQ2 from Saccharomyces cerevisiae in the defective S. pombe strain restored growth and enabled the cells to produce ubiquinone-10, indicating that COQ2 and ppt1 are functional homologs. The ppt1-deficient strain required supplementation with antioxidants, such as cysteine, glutathione, and alpha-tocopherol, to grow on minimal medium. This suggests that ubiquinone can act as an antioxidant, a premise supported by our observation that the ppt1-deficient strain is sensitive to H(2)O(2) and Cu(2+). Interestingly, we also found that the ppt1-deficient strain produced a significant amount of H(2)S, which suggests that oxidation of sulfide by ubiquinone may be an important pathway for sulfur metabolism in S. pombe. Ppt1-green fluorescent protein fusion proteins localized to the mitochondria, indicating that ubiquinone biosynthesis occurs in the mitochondria in S. pombe. Thus, analysis of the phenotypes of S. pombe strains deficient in ubiquinone production clearly demonstrates that ubiquinone has multiple functions in the cell apart from being an integral component of the electron transfer system.
| S-EPMC94818 | biostudies-literature
Project description:Ubiquinone (coenzyme Q(10) or CoQ(10)) is a lipid-soluble component of virtually all cell membranes, where it functions as a mobile electron and proton carrier. CoQ(10) deficiency is inherited as an autosomal recessive trait and has been associated with three main clinical phenotypes: a predominantly myopathic form with central nervous system involvement, an infantile encephalomyopathy with renal dysfunction, and an ataxic form with cerebellar atrophy. In two siblings of consanguineous parents with the infantile form of CoQ(10) deficiency, we identified a homozygous missense mutation in the COQ2 gene, which encodes para-hydroxybenzoate-polyprenyl transferase. The A-->G transition at nucleotide 890 changes a highly conserved tyrosine to cysteine at amino acid 297 within a predicted transmembrane domain. Radioisotope assays confirmed a severe defect of CoQ(10) biosynthesis in the fibroblasts of one patient. This mutation in COQ2 is the first molecular cause of primary CoQ(10) deficiency.
| S-EPMC1380241 | biostudies-literature
Project description:Dry bean (Phaseolus vulgaris L.) is one of the most important pulses consumed in the world. Total phenolic content, total flavonoid content, total monomeric anthocyanin content and antioxidant capacity were determined, using ferric reducing antioxidant power and free radical scavenging activity, in 255 lines grown under the same environmental conditions. For all parameters analysed, there was a wide range of variability, with differences always above one order of magnitude. Phenolic compounds in beans with coloured coats were found to be more efficient antioxidants than those with completely white coats, and samples with more strongly coloured coats (red, cream, black, pink and brown) showed the highest antioxidant capacities. Based on the strong correlation detected between the variables, total phenolic content can be considered an appropriate indicator of antioxidant activity. The results provide a robust database for selecting those lines of greater functional and nutritional interest in terms of cultivation for direct consumption, for inclusions in food formulations or for use in future breeding programs.
| S-EPMC8071416 | biostudies-literature
Project description:Endosperm extracts were prepared at various times during germination from intact castor-bean seeds and from seeds from which the embryos had been removed. The sterilized seeds were incubated either on solid water agar or on agar containing 0.3 mM-gibberellic acid. 2. Isocitrate lyase and 3-hydroxyacyl-CoA dehydrogenase had very low activities in the mature seeds, but increased 44-fold and 27-fold respectively during germination. In contrast, the extracts of mature seeds had considerable acid and alkaline lipase activity and this only increased two- to three-fold during the incubation period. 3. Incubation of the seeds with gibberellic acid accelerated the rate of appearance of isocitrate lyase and 3-hydroxyacyl-CoA dehydrogenase. It also increased the total activity attained. However, the application of hormone had, in comparison, little effect on the development of lipase activity. 4. The removal of the embryo had little influence on the development of enzyme activity in the endosperm tissue; only with isocitrate lyase was a decrease in activity observed in the absence of the embryo.
| S-EPMC1172440 | biostudies-other
Project description:BACKGROUND: Defensins are basic, cysteine-rich antimicrobial peptides that are important components of plant defense against pathogens. Previously, we isolated a defensin, PvD1, from Phaseolus vulgaris L. (common bean) seeds. RESULTS: The aim of this study was to overexpress PvD1 in a prokaryotic system, verify the biologic function of recombinant PvD1 (PvD1r) by comparing the antimicrobial activity of PvD1r to that of the natural defensin, PvD1, and use a mutant Candida albicans strain that lacks the gene for sphingolipid biosynthesis to unravel the target site of the PvD1r in C. albicans cells. The cDNA encoding PvD1, which was previously obtained, was cloned into the pET-32 EK/LIC vector, and the resulting construct was used to transform bacterial cells (Rosetta Gami 2 (DE3) pLysS) leading to recombinant protein expression. After expression had been induced, PvD1r was purified, cleaved with enterokinase and repurified by chromatographic steps. N-terminal amino acid sequencing showed that the overall process of the recombinant production of PvD1r, including cleavage with the enterokinase, was successful. Additionally, modeling revealed that PvD1r had a structure that was similar to the defensin isolated from plants. Purified PvD1 and PvD1r possessed inhibitory activity against the growth of the wild-type pathogenic yeast strain C. albicans. Both defensins, however, did not present inhibitory activity against the mutant strain of C. albicans. Antifungal assays with the wild-type C. albicans strains showed morphological changes upon observation by light microscopy following growth assays. PvD1r was coupled to FITC, and the subsequent treatment of wild type C. albicans with DAPI revealed that the labeled peptide was intracellularly localized. In the mutant strain, no intracellular labeling was detected. CONCLUSION: Our results indicate that PvD1r retains full biological activity after recombinant production, enterokinase cleavage and purification. Additionally, our results from the antimicrobial assay, the microscopic analysis and the PvD1r-FITC labeling assays corroborate each other and lead us to suggest that the target of PvD1 in C. albicans cells is the sphingolipid glucosylceramide.
| S-EPMC3996258 | biostudies-literature
Project description:Knowledge is limited about the level of bioactive compounds and antioxidant activity of seeds from bred lines of common beans developed from interspecific crosses using four different Phaseolus species (P. vulgaris L., P. coccineus L., P. acutifolius A. Gray. Gray., and P. dumosus). In this study, differences in the nutritional quality of seeds among 112 bean genotypes were evaluated by measuring the levels of phenolic compounds, pigments, antioxidant activity, and sugars. The bean genotypes were grown under high temperatures and acid soil conditions in the Amazon region of Colombia. Five typology groups of bean genotypes were identified based on the level of bioactive compounds and their functional capacity: (1) highly bioactive and functional (HBF); (2) moderately bioactive and functional (MBF); (3) moderate antioxidant content with pigment influence (MACP); (4) moderately antinutritional with limited antioxidant potential (MALAP); and (5) antinutritional, low bioactive, and functional (ALBF). We developed a nutritional quality index (NQI) with values ranging from 0 to 1 based on the nutritional and anti-nutritional balance of each genotype and the higher values of the NQI of a genotype indicating greater nutritional quality. We found three interspecific bred lines (SER 212, SER 213, and RRA 81), with NQI values higher than 0.8. These three lines belong to the typology group of HBF. The superior nutritional quality of these three interspecific bred lines is attributed to a greater level of bioactive compounds and antioxidant capacity. These three bred lines may serve as useful parents to develop nutritionally superior and stress-resilient beans from bean breeding programs. Further research is needed to explore the role of testa color in improving the nutritional quality of seeds of common bean genotypes grown under different climatic conditions.
| S-EPMC10417468 | biostudies-literature
Project description:IntroductionDespite the development of oral hypoglycemic medications, diabetes and its associated complications continue to be significant clinical issues. The purpose of this study was to examine the antidiabetic effects of Vigna radiata (L.) Wilczek seeds in mice that had been given alloxan to cause diabetes.MethodsIn Swiss albino mice, diabetes was brought on by a single intraperitoneal injection of the drug alloxan (150 mg/kg). For 14 days, glibenclamide (5 mg/kg) and methanol extract of V. radiata seeds (100, 200, and 400 mg/kg) were given orally. Following oral administration of V. radiata to mice, the blood glucose levels (BGL) and body weight were measured at 7 and 14 days. The mice were sacrificed at the end of the trial, and blood samples were taken for the evaluation of insulin, glycated hemoglobin, aspartate aminotransferase (AST), alanine aminotransferase (ALT), high-density lipoprotein (HDL), total cholesterol (TC), and triglyceride (TG) levels. It was determined how much glycogen was present in the liver. Additionally, the total phenolic and flavonoid contents of V. radiata were determined, along with the in vitro DPPH (2, 2 diphenyl-1-picrylhrazyl) free radical-scavenging activity. P < 0.05 was chosen as the cutoff for statistical significance.ResultsFollowing oral administration of V. radiata for 14 days, diabetic mice's BGL and bad cholesterol (TC and TG) levels significantly decreased, while HDL levels increased. Treatment with V. radiata significantly decreased the levels of AST, ALT, and glycated hemoglobin when compared with diabetes control. On the other hand, it raised insulin levels and the amount of liver glycogen. V. radiata underwent phytochemical analysis, which identified the presence of tannins, saponins, phenols, alkaloids, terpenoids, steroids, flavonoids, and glycosides. Per gram of V. radiata seed extract, the total phenolic content was 43.12 ± 3.14 mg of gallic acid equivalents, while the total flavonoid content was 38.35 ± 2.6 mg of quercetin equivalents. Ascorbic acid was shown to have an IC50 value of 18.64 µg/ml during a DPPH-scavenging assay, while V. radiata had an IC50 value of 73.35 µg/ml.ConclusionAccording to the findings of the current study, the methanolic extract of the seeds from the plant V. radiata possesses significant antidiabetic characteristics that are on par with those of the commonly used drug glibenclamide. Hence, V. radiata seems to be effective as a natural antidiabetic.
| S-EPMC9629934 | biostudies-literature
Project description:To dissect the gene regulatory networks operating during Scarlet Runner Bean seed development, we identified the binding sites genome-wide for transcription factor in Scarlet Runner Bean seeds during seed development using ChIP-seq
2020-07-13 | GSE153644 | GEO
Project description:Mineral deficiency is worldwide one of the major problems associated with human health, and biofortification through breeding is considered an important strategy to improve the nutritional content of staple food in countries that face this problem. The assessment of genetic variability for seed nutrient contents is a first step in the development of a biofortified crop. From the germplasm bank IDR-IAPAR-EMATER, a set of 1,512 common bean accessions, consisting of local and commercial varieties and improved lines, was analyzed. High variability among the accessions was observed for all evaluated nutrient contents (P, K, Ca, Mg, Cu, Zn, Mn, Fe and S and protein). In the mean, the contents of the carioca and black market groups (Mesoamerican gene pool), were around 7% higher for the minerals Ca, Cu, Mn and Fe and between 2-4% higher for P, K, Mg and Zn than in the other groups with Mesoamerican and Andean common bean. Few differences were observed among the Mesoamerican accessions that belong to the carioca and black commercial groups. Wide variability was observed among the evaluated genotypes, and the concentrations of the best accessions exceeded the overall mean by 14-28%. Due to the high variability in the evaluated accessions, these results may contribute to the selection of promising parents for the establishment of mating blocks. The nutritional contents of many of the improved lines evaluated in this study were higher than those of the commercial cultivars, indicating the possibility of developing new biofortified cultivars.
| S-EPMC7521705 | biostudies-literature
Project description:We conducted a survey of fungal endophytes in 582 germinated seeds belonging to 11 Colombian cultivars of the common bean (Phaseolus vulgaris). The survey yielded 394 endophytic isolates belonging to 42 taxa, as identified by sequence analysis of the ribosomal DNA internal transcribed spacer (ITS) region. Aureobasidium pullulans was the dominant endophyte, isolated from 46.7 % of the samples. Also common were Fusarium oxysporum, Xylaria sp., and Cladosporium cladosporioides, but found in only 13.4 %, 11.7 %, and 7.6 % of seedlings, respectively. Endophytic colonization differed significantly among common bean cultivars and seedling parts, with the highest colonization occurring in the first true leaves of the seedlings.
| S-EPMC4857701 | biostudies-literature