Characterization of Blighia sapida seed extracts.

Blighia sapida, commonly known as the ackee, is a member of the Sapindaceae family. The tree is native to the forests of West Africa and was brought to the Caribbean and later Florida, where it is cultivated as an orchard crop in some areas. Arilli of the fruit are processed to make canned ackee in brine whereas the seeds, raphe and pods are discarded. Physiochemical studies were conducted on extracts of the seed. Qualitative analysis detected the presence of phenolics and reducing sugars. Aqueous extracts of the seeds (ASE) exhibited free radical scavenging activity and had an inhibitory concentration of 2.59 mg/mL. Gas chromatography mass spectrometry led to the identification of several metabolites including amino acids and fatty acids. Hypoglycin B was isolated utilizing ion exchange chromatography. Fourier transform infrared spectroscopy of hypoglycin B detected a band resonating at 3070 cm-1 which may be attributed to the methylenecyclopropane moiety of hypoglycin B. The seeds had a lipid content of 5.72 ± 0.25 % (w/w). The ackee seed oil (ASO) had a saponification value of 152.07 ± 37 and a carotenoid content of 23.7 ± 1.8 mg/kg. The ackee seeds are a source of bioactive components.

Changes in secondary metabolites and fiber quality of cotton (Gossypium hirsutum) seed under consecutive water stress and in silico analysis of cellulose synthase and xyloglucan endotransglucosylase.

Global warming has led to severe drought conditions. The selection of plant varieties that can withstand drought and produce increased yields are of utmost importance. In the current study, secondary metabolites, seed trait and fiber characteristic of cottonseeds (Gossypium hirsutum) exposed to double and third water stress exposure was investigated. Total phenol and tannin content in W1S33 increased significantly after third water stress exposure. Accumulation of wax was enhanced in seeds of W3S33 and W3S34 that were subjected to third water stress. Fiber quality parameters decreased when cottonseeds were rainfed. High irrigation resulted in fragile and delicate fiber. Seeds grown under 66% FC irrigation saved water and produced seeds that had the potential of producing high quality fibers. In silico analysis was performed on cellulose synthase A (CesA) and xyloglucan endotransglycosylase (XET) enzymes present in Gossypium hirsutum. The intracellular locations of the CesA and XET1 enzymes are the plasma membrane and cell wall, respectively. Proline is conserved in the C-terminal of the CesA enzyme and plays an important role in enzyme functionality. This study provides a better understanding as to the mechanisms by which the plant can tolerate and combat water stress conditions as well as reduce water consumption. In order to grow cotton seeds with desirable morphometric characteristics and optimal fibers under water stress exposure and in dry areas, it is better to use seeds that are irrigated under optimal irrigation conditions, ie 66% FC.

Anatomic features, tolerance index, secondary metabolites and protein content of chickpea (Cicer arietinum) seedlings under cadmium induction and identification of PCS and FC genes.

ABSTRACT: Chickpea (Cicer arietinum) belonging to the Fabaceae family is a major legume crop and is a good source of protein and carbohydrates. Industrialization has resulted in soil contamination with heavy metals such as cadmium. Adsorption of cadmium by plants can lead to reduced yields and heavy metal toxicity. In the current study, changes in the anatomical, morphological features and biochemical properties of the chickpea plant were evaluated. Two indexes DWSTI and PHSTI were determined. Anatomically, there was a change in the number of xylem poles within the root structure which was most significant at treatments of 125 µg cadmium. There was also a noticeable change in leaf pigmentation, the total phenolics and soluble protein in the plant. Cadmium levels were elevated attaining concentrations of 0.21, 0.40 and 0.52 mg per gram dry weight in plants exposed to 62, 125 and 250 µg/g Perlit cadmium after a period of 30 days. A noticeable increase in the level of cadmium in the plant was observed. Two PCS genes, glutathione gamma-glutamylcysteinyltransferase 1 and glutathione gamma-glutamylcysteinyltransferase and four FC genes, 4 proteins and 4 mRNA were detected in chickpeas. Bioinformatics tools were utilized to predict enzyme structure and binding sites. Chickpea may be classified as a cadmium hyperaccumulator and may be considered for use in phytoremediation. This study provides a better understanding with regards to the response of chickpeas to cadmium and the genetic mechanism by which the plant regulates heavy metal toxicity.

In silico prediction, phylogenetic and bioinformatic analysis of SoPCS gene, survey of its protein characterization and gene expression in response to cadmium in Saccharum officinarum.

Phytochelatin synthase isolated from microorganisms, yeasts, algae and plant, serve a fundamental role in reducing heavy metals. In this research the in silico PCS gene structure (SoPCS) of sugarcane, its secondary and 3D protein structure, physicochemical properties, cell localization and phylogenetic tree were predicted utilizing bioinformatics tools. SoPCS expression in the leaves and roots of sugarcane in tissue culture treated with cadmium was also studied utilizing real time PCR. The predicted SoPCS gene contains 1524 nucleotides, a protein encoded with 508 amino acids of which the molecular weight is 55953.3 Da, 6 exons and 5 introns. The subcellular position of the enzyme is mitochondrion or cytoplasmic. Two domains belonging to the phytochelatin synthase family with similar features was found in Pfam having more than 97% similarity with the predicted SoPCS protein. Phylogeny analyses of plant species were well isolated from other organisms. Ten disulfide-bonded cysteines were excluded from the structure of SoPCS. The predicted 3D structure of SoPCS showed that it is able to bind to L-gamma-glutamylcysteine as substrate. The binding site sequence of PCS included amino acids 52(Q),55(P),56(A),57(F), 58(C),103(G),104(I),151(S),163(G),165(F),206(D), 213(R). The common amino acid with conserved sequence in the binding site of the plant was 103Gly. Gene expression indicated that SoPCS has an important role in the response of sugarcane to cadmium with potential use in genetic engineering to remove metal contaminants in the environment. This is the first characterization of a PCS from sugarcane.

Characterization of Jamaican Delonix regia and Cassia fistula Seed Extracts.

Delonix regia and Cassia fistula seed extracts were evaluated for their antioxidant activity, total phenolics, ash, zinc and fatty acid content. Fourier Transform Infrared Spectroscopy (FTIR) was utilized to assess the chemical functionalities present within the seeds. Antioxidant activity was determined by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Trolox equivalent antioxidant capacity (TEAC) assays. Total phenolics were determined by the Folin-Ciocalteu assay. Lipid extracts were characterized by nuclear magnetic resonance spectroscopy and gas chromatography/mass spectrometry. Zinc concentration was determined by atomic absorption spectroscopy. Extracts from the seeds of C. fistula had a higher antioxidant activity, free radical scavenging activity, and phenolic content than D. regia. FTIR revealed that the seeds are a rich source of protein with small quantities of fat. C. fistula extracts contained a higher percentage of total fat than D. regia. Palmitic acid was identified as the predominant saturated fatty acid in both extracts. Oleic acid and linoleic acid were identified in smaller quantities. Seed extracts may be considered for use in food and nutraceutical applications.

Purification and Characterization of Phenylalanine Ammonia Lyase from Trichosporon cutaneum.

Trichosporon cutaneum phenylalanine ammonia lyase was selected as a model to investigate the dual substrate activity of this family of enzymes. Sequencing of the PAL gene identified an extensive intron region at the N-terminus. Five amino acid residues differing from a prior report were identified. Highest Phe : Tyr activities (1.6 ± 0.3 : 0.4 ± 0.1 µ mol/h g wet weight) were induced by Tyr. The enzyme has a temperature optimum of 32°C and a pH optimum of 8-8.5 and shows no metal cofactor dependence. Michaelis-Menten kinetics (Phe, K m 5.0 ± 1.1 mM) and positive allostery (Tyr, K' 2.4 ± 0.6 mM, Hill coefficient 1.9 ± 0.5) were observed. Anion exchange chromatography gave a purification fold of 50 with 20% yield. The His-Gln motif (substrate selectivity switch region) indicates the enzyme's ability to act on both substrates.