Effect of incubation period on the glycosylated protein content in germinated and ungerminated seeds of mung bean (Vigna radiata (L.) Wilczek).

The effects of different incubation periods on the contents of amino acids, proteins, glycosylated proteins and metabolites in germinated and ungerminated mung bean seeds were investigated in this study. The study employs soaking of mung bean seeds in water under laboratory conditions at 28 °C for 3, 6, and 9 h, followed by germination for 12, 24, 36, and 48 h. Seeds collected from different period of imbibition and germination were subjected to total protein extraction for phytochemical analysis. Germination of the seeds was found to be most successful after 6 h of soaking (rather than 9 h of incubation). Hence, seeds imbibed for 6 h were further investigated for germination at 28 °C for 12, 24, 36, and 48 h. Total protein was extracted from both imbibed and germinated seeds, followed by trypsin digestion. Liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based peptide mass fingerprinting revealed 38 proteins in 6 h water-imbibed seeds and 50 proteins in 24 h germinated seeds. Among these, 16 were identified as glycosylated proteins and the maximum number of glycosylated proteins were detected in 6 h water-imbibed seeds and 24 h germinated seeds. Moreover, High Performance Liquid Chromatography (HPLC) was used to quantify amino acids from the extracted proteins. A total of 15 amino acids were detected, of which eight were essential and the remaining were non-essential; amino acid concentrations increased following 3, 6, and 9 h of imbibition when compared to the control. It was concluded from the study that seeds with 6 h of imbibition and 24 h of germination can be used as potential nutritional source of different amino acids, proteins, glycosylated proteins, and other bioactive metabolites in human diet.

Synthesis of a 1,2,3-bistriazole derivative of embelin and evaluation of its effect on high-fat diet fed-streptozotocin-induced type 2 diabetes in rats and molecular docking studies.

The embelin derivative 2a was synthesized with the 1,2,3-bistriazole and spectral data confirmed its structural identity. Anti-diabetic and anti-lipidemic effects were evaluated using HFD-STZ induced type 2 diabetic rats. The derivative 2a (30 mg/kg b wt.) supplementation significantly (P ≤ 0.01) normalized the changed biochemical parameters like fasting blood glucose (FBG), body weights, plasma insulin level, total cholesterol (TC), triglycerides (TG) and marker enzymes of carbohydrate metabolism. The derivative 2a (30 mg/kg) also showed a significant effect on oral glucose tolerance test (OGTT) and intraperitoneal insulin tolerance test (ITT). But 15 mg/kg dose of derivative 2a failed to show any significant effects in HFD-STZ induced type2 diabetic rats. Histopathology analysis substantiated the protective effect of this derivative 2a (30 mg/kg b wt.) on the ß-cells of the pancreatic, liver and adipose tissues in diabetic treated rats. Further, the expressions of PPARγ and GLUT4 were significantly enhanced in the epididymal adipose tissue. The HOMO and LUMO energies characterized the molecular stability of the derivative 2a with 6-311G++ (d, p) in DFT/B3LYP/LanL2DZ method using Gaussian09 program package. The molecular docking analysis also confirmed the activity of derivative 2a through hydrogen bond interaction with ARG 288, GLU 343, SER 342 and least energy value (-7.72 kcal/mol). Hence, the embelin-1,2,3-bis triazole derivative 2a (30 mg/kg) enhanced the activity of embelin and might be acting as a suitable drug for type 2 diabetes, obesity and its complications.

Indigenous plant growth-promoting bacteria enhance plant growth, biomass, and nutrient uptake in degraded forest plants.

The present study was aimed to isolate indigenous plant growth-promoting bacteria (PGPB) from Nanmangalam reserve forest, India and to analyze their positive impact on nursery plant species. In total, 160 isolates were obtained from different nitrogen-free Media (LGI, JMV, NFB). Amongst these, 12 isolates were shown positive for 5-8% of ammonia production nif H positive and then isolates were further tested for their plant growth-promoting (PGP) activity. Based on their PGP activity, nine isolates were selected, and applied in nurseries of twelve native plant species, along with organic manure and inorganic fertilizer. All the isolates were shown positive effects when compared to control. In that, five of these bacterial isolates, Paenibacillus sp. RRB2, Azospirillum brasilense RRAK5, Bacillus subtilis subsp. subtilis RRD8, Burholdria kururiensis RRAK1, and Pseudomonas stutzeri RRAN2, enhanced biomass production in several trees.

Bioactive potential and composition analysis of sulfated polysaccharide from Acanthophora spicifera (Vahl) Borgeson.

Marine seaweeds contain a valuable source of functional bioactive polysaccharide and it plays main role for effective anticancer activity. The structural feature of SPs was studied through FT-IR and 1H NMR spectra analysis. The isolated SPs from A. spicifera contain 63.3% of total sugar, 21.9% of total sulfate and 12.6% of total uranic acid was found. The active F2 fraction molecular weight of SP was found to be 420 kDa. The sugar was composed of galactose (73.5%), xylose (9.2%), mannose (1.9%) and arabinose (10.9%). Further the SP showed DPPH free radical scavenging activity of 55.55% at 150 µg/mL and reducing power activity of 91.3% at 125 µg/mL. In the present study, the purified sulfated polysaccharide (fraction F2) were extracted, purified and characterized for red seaweed and evaluated for their potential anticancer activity of in A549 cell lines under in vitro condition. These polysaccharide fractions exhibited potential apoptotic effects on A549 cell lines.

Mass cultivation of UV-B adapted Arthrospira platensis RRGK under open raceway pond for the production of Poly-ß-hydroxy butyrate.

Six different strains of cyanobacteria were isolated from the freshwater lake, Arakkonam, India. Staining of cells with Nile Red showed the presence of large quantities of PHB granules in the cell cytoplasm of Arthrospira. Molecular identification of the strain was carried out using 16S rRNA analysis and their systematic position was ascertained as Arthrospira platensis RRGK. Studies were carried out on random mutagenesis approach using UV-B radiation for enhancing the production of PHB. Further, Response Surface Methodology was used for optimization of pH, temperature, and sodium bicarbonate for higher biomass and PHB production. Under open raceway pond A. platensis RRGK produced biomass concentration of 2.2±0.13gL-1 and 131±0.36gL-1 of PHB content. It was chemically characterized through FTIR, DSC, TGA and XRD analyses. Hence, PHB can be produced from cyanobacteria by sequestering harmful CO2. It can also be used as a substitute for synthetic polymers in tissue engineering.

Optimization of polyhydroxybutyrate production utilizing waste water as nutrient source by Botryococcus braunii Kütz using response surface methodology.

Investigations have been made to optimize various factors including pH, temperature, and substrate for enhanced polyhydroxybutyrate (PHB) production in Botryococcus braunii which serves as a pioneer for production of bioplastic (PHB). Polyhydroxybutyrate is a natural, decomposable polymers accumulated by the microorganism under different nutritional condition. Strain selection was done by staining method using Sudan black and Nile red dye. Using response surface methodology (RSM), three level- three variables Box Behnken design (BBD), the best potential combination of pH (4-11), temperature (30-50°C) and sewage waste water as substrate fed at different concentrations at 20%-100% for maximum PHB production was investigated. Maximum yield (247±0.42mg/L) of PHB dry weight was achieved from the 60% concentration of sewage waste water as a growth medium at pH 7.5 at 40°C. It was well in close agreement with the value predicted by RSM model yield (246± 0.32mg/L). Thus the study shows the production of PHB by B. braunii along with the basic characterization of PHB by using FTIR and TEM analysis. These preliminary studies indicated that PHB can also be produced by B. braunii utilizing waste water. There is no report on the optimization of PHB production in this microalgae have been documented.

Biosynthesis, purification and characterization of polyhydroxybutyrate from Botryococcus braunii kütz.

Polyhydroxybutyrate (PHB) is completely biodegradable which is metabolised by microorganisms in the soil as their sole food source in few years. The level of PHB up to 10.6% of algal dry weight is of great potential of the eco-friendly nature. Botryococcus braunii is mainly used for the production of biodiesel and is also capable of producing biopolymer polyhydroxy butyrate (PHB). In this study, Botryococcus braunii is used which generally produce PHB to around 20% of the dry weight. Three different microalgae were isolated from the fresh water of Kolavoi lake of Tamil Nadu. They were identified by their morphological features under the light microscope. The primary screening of PHB intracellular granules was made by using Nile red dye under a fluorescent microscope. Among them, Botryococcus braunii showed high accumulation of PHB granules. For authentic confirmation, the chloroform extracted PHB was analysed by FTIR, XRD and DSC-TGA analyses to characterize PHB with commercial biodegradable thermoplastic. This is the first report in B. braunii for its PHB production.

Isolation, purification and characterization of antimicrobial protein from seedlings of Bauhinia purpurea L.

A novel antimicrobial protein was purified from the seedlings of Bauhinia purpurea by sequential procedures entailing ammonium sulfate precipitation, cation exchange chromatography, preparative native-PAGE and a yield of 2.7% was obtained from the crude extract. The purified antimicrobial protein appeared as a single protein band on SDS-PAGE with the molecular mass of 20.9 kDa. Purified antimicrobial protein exhibited a potent antimicrobial activity against both Gram-positive and Gram-negative bacteria. Analysis of the trypsin digested peptides of purified protein using the MALDI-TOF MS/MS resulted in the identification of 174 amino acids. The purified protein had an optimum of pH of 5.5 and was stable at 35 °C for exhibiting its maximal antibacterial activity. The addition of metal ions such as Mn(2+) and Ca(2+) to the purified protein enhanced the antimicrobial activity of purified protein. The MIC of purified protein against Bacillus cereus and Escherichia coli were 13 µg/ml and 15 µg/ml, respectively. The purified protein digested the peptidoglycan layer of bacteria which was visualized by TEM analysis.

Stabilization of mitochondrial and microsomal function of fucoidan from Sargassum plagiophyllum in diethylnitrosamine induced hepatocarcinogenesis.

Crude fucoidan from Sargassum plagiophyllum extracted from blade and purified by Q-Sepharose fast flow anion-exchange chromatography and three fucoidan fractions were obtained. Maximum sulphate containing fucoidan fraction was considered as purified fucoidan and purity was checked with agarose gel electrophoresis. The monosaccharides of purified fucoidan analysed by HPLC revealed the presence of the sugars such as fucose as a major sugar were 70.8 mol%. The percentages of other sugars were galactose (13.5%), xylose (2.5%) and mannose (11.2%). GPC was used to analyse molecular weight of purified fucoidan and it was found to be 35 kDa. The levels of ICDH, SDH, MDH, a-KGDH, Phase-I biotransformation enzymes, and Phase-II biotransformation enzymes were decreased in cancer bearing animals which may be due to oxidative stress and mitochondrial damage and fucoidan restored these enzyme activities. The inhibition of carcinogen metabolic activation indicates the anticancer activity of fucoidan in DEN induced liver cancer.

Studies on screening, isolation and purification of a fibrinolytic protease from an isolate (VK12) of Ganoderma lucidum and evaluation of its antithrombotic activity.

Antithrombotic activity of a protease purified from a medicinal mushroom, Ganoderma lucidum, has been evaluated platelet aggregation in vitro and pulmonary thrombosis in vivo. The purified protease exhibited concentration dependent inhibitory effects on platelet aggregation induced by ADP (adenosine diphosphate), with an IC(50) value of 2.4 mg/mL. The purified protease protected mice against thrombotic death or paralysis induced by collagen and epinephrine in a dose dependent manner when administered orally. It produced a significant inhibition of thrombotic death or paralysis at 60 µg/kg body weight, while aspirin produced a significant inhibition of thrombosis at 10-20 mg/kg body weight. The purified protease also has showed fibrinolytic activity and alters coagulation parameters such as activated partial thromboplastin time (APTT), and thrombin time (TT) in rat platelet. These results suggested that the antithrombotic activity of Ganoderma lucidum protease might be due to antiplatelet activity rather than anticoagulation activity.