In plants, it is well-known that ascorbic acid (vitamin C) can be synthesized via multiple metabolic pathways but there is still much to be learnt concerning their integration and control mechanisms. Furthermore, the structural biology of the component enzymes has been poorly exploited. Here we describe the first crystal structure for an L-galactose dehydrogenase (SoGDH from spinach), from the D-mannose/L-galactose (Smirnoff Wheeler) pathway which converts L-galactose into L-galactono-1,4-lactone. The kinetic parameters for the enzyme are similar to those from its homologue from camu-camu, a super-accumulator of vitamin C found in the Peruvian amazon. Both enzymes are monomers in solution, have a pH optimum of 7 and their activity is largely unaffected by high concentrations of ascorbic acid, suggesting the absence of a feedback mechanism acting via GDH. Previous reports may have been influenced by changes of the pH of the reaction medium as a function of ascorbic acid concentration. The structure of SoGDH is dominated by a (β/α)8 barrel closely related to aldehyde-keto reductases (AKRs). The structure bound to NAD+ shows that the lack of Arg279 justifies its preference for NAD+ over NADP+, as employed by many AKRs. This favours the oxidation reaction which ultimately leads to ascorbic acid accumulation. When compared with other AKRs, residue substitutions at the C-terminal end of the barrel (Tyr185, Tyr61, Ser59 and Asp128) can be identified to be likely determinants of substrate specificity. The present work contributes towards a more comprehensive understanding of structure-function relationships in the enzymes involved in vitamin C synthesis.
Septins possess a conserved guanine nucleotide‐binding (G) domain that participates in the stabilization of organized hetero‐oligomeric complexes which assemble into filaments, rings and network‐like structures. The fruit fly, Drosophila melanogaster, has five such septin genes encoding Sep1, Sep2, Sep4, Sep5 and Pnut. Here, we report the crystal structure of the heterodimer formed between the G‐domains of Sep1 and Sep2, the first from an insect to be described to date. A G‐interface stabilizes the dimer (in agreement with the expected arrangement for the Drosophila hexameric particle) and this bears significant resemblance to its human counterparts, even down to the level of individual amino acid interactions. On the other hand, a model for the G‐interface formed between the two copies of Pnut which occupy the centre of the hexamer, shows important structural differences, including the loss of a highly favourable bifurcated salt‐bridge network. Whereas wild‐type Pnut purifies as a monomer, the reintroduction of the salt‐bridge network results in stabilizing the dimeric interface in solution as shown by size exclusion chromatography and thermal stability measurements. Adaptive steered molecular dynamics reveals an unzipping mechanism for dimer dissociation which initiates at a point of electrostatic repulsion within the switch II region. Overall, the data contribute to a better understanding of the molecular interactions involved in septin assembly/disassembly.
Rosewood, Aniba rosaeodora is an endangered species in Amazon forests and its natural stands have been heavily depleted due to over-exploitation for the cosmetic industry. This study aimed to investigate the genetic diversity and population structure of 90 rosewood accessions from eight localities in the Peruvian Amazon through 11 Inter Simple Sequence Repeats (ISSR) primers. The ISSR primers produced a sum of 378 bands, of which 375 (99.2%) were polymorphic, with an average polymorphism information content (PIC) value of 0.774. The mean effective number of alleles (Ne), Shannon informative index (I), gene diversity (He) and total gene diversity (Ht) were 1.485, 0.294, 0.453 and 0.252, respectively. Analysis of molecular variance (AMOVA) showed the presence of maximum variability within populations (88%). The Structure algorithm, neighbor joining and principal coordinate analysis (PCoA) grouped the 90 rosewood accessions into three main populations (A, B and C). Diversity indices at the inter-population level revealed a greater genetic diversity in population A, due to higher gene flow. The neighbor-joining analysis grouped populations A and B, while population C was found to be divergent at the inter population level. We concluded that population A reflects higher genetic diversity and should be prioritized for future management and conservation plans.
Environmental pollution is a severe and common problem in all the countries worldwide. Various physicochemical technologies and organisms (e.g., plants, microorganisms, etc.) are used to address these environmental issues, but low-cost, practical, efficient, and effective approaches have not been available yet. Microalgae offer an attractive, novel, and little-explored bioremediation alternative because these photosynthetic organisms can eliminate pathogenic microorganisms and remove heavy metals and toxic organic compounds through processes still under study. Our research team has conducted some experiments to determine the bioremediation potential of native microalgae on some pollutant sources (i.e., leachate and wastewater) and its ability to remove hazardous chemical compounds. Therefore, in this chapter, we provide the results of our research and updated information about this exciting topic. Experiments were conducted under controlled culture conditions using several native microalgae species, variable time periods, different pollutant sources, and hazardous chemicals such as ethidium bromide. The results indicated that native microalgae can remove pollutants (i.e., phosphorus, ammonia, etc.) of wastewater, leachate, and some hazardous chemical compounds such as ethidium bromide. In conclusion, native microalgae have an excellent potential for removing several pollutants and, consequently, could be used to develop bioremediation technologies based on native microalgae from the Peruvian Amazon.
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