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The Stability of protease in organic solvent media has been widely discussed for more than two decades. Proteases can catalyze synthetic reactions in organic media, by this way solvent stabilities of proteases are very important. In this study, we reported a bacterium isolated from hot spring of Geinarje, Iran producing an organic solvent stable protease. Protease producing bacteria were screened on skim milk agar and the formation of a clear zone around the bacterial colony was investigated. Proteolytic activity was assayed by a modified caseinolytic method using casein as a substrate. The best alkaline protease producing bacterium was selected and identified on the basis of 16S rDNA gene sequencing and morphological and biochemical characteristics. The effect of organic solvents, temperature, pH, and NaCl on proteolytic activity were examined. According to phylogenetic analysis, morphological and physiological tests, isolated, the bacterium was identified as a new strain of Brevibacillus borstelensis. This strain was able to produce an extracellular organic solvent-stable protease with 0.53U/ml enzyme activity. After 2 hour incubation at 30°C the protease of Brevibacillus borstelensis AMN was active in wide ranges of organic solvents, and its activity was enhanced in the presence of 25% (V/V) isopropanol. The biochemical properties of the enzyme revealed that the optimal pH and temperature for protease activity were 9.0 and 60°C, respectively. Our finding indicated that these robust properties of protease, like outstanding activity and stability in organic solvents and alkaline medium, might be applicable for various industrial biotechnologies.
 

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Objective: Ischemic preconditioning (IPC) is an endogenous phenomenon that can induce ischemic tolerance (IT) in variety of organs such as brain. In this study, we examined the intermittent and prolonged normobaric hyperoxia (HO) on neurologic deficit scores, infarct volume, and superoxide dismutase activity. Materials and Methods: The rats were divided to four main groups. First two main groups were exposed with HO in prolonged (24 h; PrHO) and intermittent (4 h×6 days; InHO) groups and second two main group acted as controls, and were exposed to 21% oxygen in the same chamber (room air, RA) continuously (24 h; PrRA) and discontinuously (4 h×6 days; InRA). Each group subdivided to three subgroups. After 24 h, first subgroup were subjected to 60 minutes MCAO followed by 24 h of reperfusion. Then, IT induced by InHO and PrHO were measured by neurologic deficit scores and infarct volume. Second and third subgroups were called sham-operated and intact subgroups for assessment of the effect of HO on superoxide dismutase activity. Results: Our findings indicate that InHO and PrHO are involved in the induction of IT. Pretreatment with InHO and PrHO reduced neurologic deficit scores and infarct volume significantly. InHO and PrHO increase superoxide dismutase activity significantly. Conclusion: Although further studies are needed to clarify the mechanisms of ischemic tolerance, InHO and PrHO seem to partly exert their effects via increase superoxide dismutase activity.

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    In this study to determine the effect of sonication time and intensity on the activity of the alpha-amylase, Aspergillus Oryzae PTCC 5164 as a Producer of enzyme was used in the solid staste fermentation system.  A combination of wheat and barley (0:100, 50:50, 100:0) as a substrate, Ultrasound with the intensity treatment (35, 5/67 and 100%) and 5, 10 and 15 minutes were used. Statistical analysis with software Design Expert 6.0.2 showed that the effect of ultrasound on enzyme produced in culture platform is different. Alpha-amylase production with barley as the substrate had more resistant during the sonication. Also by increasing the time and intensity of sonication, negative effects of cavition became more apparent and enzyme activity decreased. Diminishing activity of the alpha-amylase produced, depends on the duration and intensity of sonication and combination of solid culture. Probably the π₀ → π* amide transitions and secondary structural components, especially b-sheet, of the enzyme was significantly influenced by ultrasound.

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Effects of yeast extract (YE) (0, 0.05, 0.1, 0.2, 0.5 and 1%) were investigated on rosmarinic acid (RA) accumulation and tyrosine aminotransferase (TAT) gene expression in Melissa officinalis seedlings at different time intervals (4 and 17 hours). Based on the results, only YE concentrations of 0.05%, 0.1%, and 0.2%, for the 17-hour treatment, significantly stimulated RA biosynthesis pathway. At this elicitation time, flavonoid content and TAT gene expression significantly increased by the increase in YE concentration up to 0.2% as compared to the control. The maximum amount for both of them was seen at 0.1% YE treatment, where the RA accumulation was drastically elevated. Furthermore, the aerial parts were improved when root lengths were decreased; this was observed in seedlings that were treated by YE for 30 days. Overall, these observations can be attributed to the oxidative stress induced by YE, as a consequence of its uptake by the plant, as revealed by increasing activities in superoxide dismutase and catalase.

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Application of chemical pesticides has increased significantly worldwide and has raised serious concerns about environmental pollutions. One of the encouraging trends to minimize pesticide risk is production of resistant plants containing toxic proteins against insect pests. Considering the importance of purification and characterization of digestive enzymes in the production of resistant plants, in this study an α-glucosidase from the Naranga aenescens Moore's midgut was purified by ammonium sulfate precipitation, ion exchange chromatography on DEAE-sepharose, and concentrating through ultrafiltration. The apparent molecular mass of the enzyme was 48 kDa determined by SDS-PAGE. The optimum pH and temperature of the enzyme were 6.0 and 45°C, respectively. The irreversible thermoinactivation of the enzyme showed that it was highly stable at 35ºC but moderately stable at 40 and 45ºC. Zn²⁺, Hg²⁺, Co²⁺ at 10 and 20 mM, and Ba⁺²only in 20 mM strongly inhibited the α-glucosidase activity. Ba²⁺ and Ca²⁺ only at 10 mM, EDTA and Hg₂²⁺ only at 20 mM and Mg²⁺ at 10 and 20 mM significantly increased the enzyme activity. The K_m and K_cat values for the α-glucosidase were 0.54 mM and 3.62 min^-1, respectively, when p-Nitrophenyl-α-D-glucopyranoside (pNαG) was used as a substrate.

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Lipases are known to have important functions in many physiological processes in plants. Here, we cloned a lipase gene via Rapid Amplification of cDNA Ends (RACE) technique from Brassica napus L., designated as BnDIL1 (B. napus Desiccation-Induced Lipase 1). The lipase enzyme activity was confirmed by estimating the lipase activity and reduced lipids content in Saccharomyces cerevisiae (pep4)transformant. Two B. napus lines with different oil contents were employed to examine the transcription profiles of BnDIL1 during the processes of seed morphogenesis, maturation, dormancy, pregermination and germination. The transcription level of lipid degradation pathway was enhanced during the processes of seed maturation, dormancy, pregermination and germination, and was higher in seeds of low oil-contents line than that of high oil-contents line. However, BnDIL1 was significantly activated when seed desiccation started. Both “slow desiccation” and “fast desiccation” treatments on seedlings dramatically activated the transcription of BnDIL1, while only “slow desiccation” stress, which would induce the cell apoptosis, significantly activated the transcription of lipid degradation gene. This result demonstrated that BnDIL1 in B. napus was desiccation stress dependent gene rather than fatty acids degradation gene.

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In 2021, an experiment was conducted to study the quality and storage of ‘Sena’ variety of tomato by application of calcium oxide, gamma-aminobutyric acid and salicylic acid in the postharvest laboratory of department of horticultural sciences and engineering in Vali-e-Asr university of Rafsanjan. The statistical design of the experiment was a factorial in the form of a completely random design. The first factor included control (distilled water), 0.5% calcium oxide (CaO), gammaaminobutyric acid with 10 mM concentration (GABA), salicylic acid with a concentration of 0.5 mM (SA) and combined treatment (CaO + GABA + SA), and the second factor included 3 storage times (7, 14 and 21 days). The results of the experiment showed that the effect of the combined treatment was significant in maintaining the quantitative and qualitative characteristics of tomatoes. The lowest content of soluble solids, the lowest activity percentage of pectin methyl esterase enzyme, the highest amount of phenol and the highest water activity were observed in gamma-aminobutyric acid treatment. Compared to the control sample, salicylic acid treatment prevented up to 32% of ion leakage, and 2.5% maintained the weight, and maintained titratable acidity on 7th day comparing the first day. According to the results, the highest level of firmness, general acceptance, water activity and vitamin C belonged to calcium oxide treatment. The lowest amount of malondialdehyde and lycopene, the lowest level of polygalacturonase enzyme activity (0.09% reduction in activity compared to the control), the lowest microbial contamination (0.63% colony reduction compared to the control), the lowest percentage of ion leakage and frostbite belonged to calcium oxide treatment. In general, among the treatments, 0.5% calcium oxide and 10 mM gamma-aminobutyric acid treatments best maintained the quantitative and qualitative traits of tomato fruit and are suggested for its storage.
 

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Chicory (Cichorium intybus L.) is a typical Mediterranean plant distributed throughout the world and has different commercial uses such as salad, forage, inulin production, and coffee substitute. Health promoting characteristics of inulin as a prebiotic compound led to its biosynthesis pathway discovery. Two enzymes, namely, 1-SST and 1-FFT, are involved in inulin biosynthesis during normal phase. By cold nights or other factors, 1-FEHs enzymes degrade inulin to fructosyl units. To compare the strength of function of these genes in a wild type genotype with root type cultivar (Orchies) of chicory at three stages, i.e. 60, 90, and 120 days after seed planting, relative expression of those genes along with their corresponding metabolites were assessed using RT-qPCR and HPLC. Expression results showed that, unlike Orchies cultivar, relative expression of 1-SST in wild type genotype was ascending, relative expression of 1-FFT was very low and constant and there were high levels of relative expression of 1-FEH I gene during growing season due to flowering initiation. Also, glucose and fructose concentrations were upward, as result of 1-SST and 1-FEH I enzymes activity in wild type genotype, respectively. Degree of Polymerization (DP) of produced inulin had almost no increase due to low function of 1-FFT enzyme in the wild type genotype (DP< 5), but Orchies cultivar produced inulin with DP> 10 as expression of 1-SST decreased during growing season. So, it is possible to make inulin pathway in root type chicory cultivars more efficient by expanding and overexpressing 1-SST function using such wild resources through backcross breeding or biotechnology methods.
 

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In this study, biochemical responses of three common bean genotypes were investigated under three levels of water deficit (75, 50 and 25% FC) at pre- flowering stage. The results showed different effects of drought stress on photosynthetic pigments, total carotenoids, Electrolyte Leakage (EL), Relative Water Content (RWC), lipoxygenase enzyme activity, lipid peroxidation (MDA), total soluble proteins, proline and Abscisic Acid (ABA) content in K-S-31167, GE-288 and NAZ genotypes. Our results showed different patterns in antioxidant enzymes activity including Catalase (CAT), Guaicol PerOxidase (GPOX) and Ascorbate PerOxidase (APX) in K-S-31167 as drought tolerant, GE-288 and NAZ as semi tolerant and sensitive genotypes, respectively. Results showed that drought stress response in common beans is highly genotype dependent.
 

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Waterlogging and drought are severe constrains that limit maize seedling growth in tropical and subtropical regions. It is significant to determine the differences in morphological and physiological responses of maize to drought and excess soil water, with a view toward better breeding and field management. In the present experiment, different levels of soil water availability were initiated at the one-leaf (V1) stage of two maize cultivars (Denghai9 and Yidan629): Control (CK), Severe Drought (SD), Light Drought (LD), Severe Waterlogging (SW), and Light Waterlogging (LW). The results indicated that waterlogging had more discernible impact on the seedling growth of both cultivars than drought stress. The Relative Growth Rate (RGR) of shoots and roots, along with root length, volume, and surface area were all markedly decreased in both cultivars under waterlogging stress. The malondialdehyde content increased significantly in roots and leaves under waterlogging treatment. In both cultivars, SuperOxide Dismutase (SOD) was mostly activated in roots and leaves at the three-leaf (V3) stage by waterlogging stress, while the Catalase (CAT) activity apparently increased under drought stress. The activity of Peroxidase (POD) distinctly enhanced in both cultivars under drought and waterlogging stress. Ascorbate Peroxidase (APX) showed constant activity with prolongation of waterlogging stress, and Glutathione Reductase (GR) activity notably increased in roots under waterlogging conditions at the six-leaf (V6) stage. We concluded that SOD, POD, APX, and GR were the most important antioxidant enzymes under waterlogging conditions, whereas CAT and POD appeared to play key roles under drought stress.

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The tomato leafminer (Tuta absoluta Meyrick) (Lepidoptera: Gelechiidae) is one of the most important pests of tomato worldwide. In this study, resistance of different populations of the tomato leaf miner from Isfahan Province was evaluated against abamectin. The median Lethal Concentrations (LC₅₀) of different populations were estimated by bioassays using a leaf-dip method. The LC₅₀ value of abamectin in the reference population of Isfahan University of Technology (IUT) was estimated as 5.67 mg ai L^-1, while the population of Shahre-e-Abrisham 1 showed the highest (25-fold) resistance, with an LC₅₀ value of 143.18 mg ai L^-1. Pre-treatment of different populations with diethyl maleate (DEM) synergist, an inhibitor of glutathione-S-transferases (GSTs), increased significantly abamectin toxicity. GST activity was also found significantly different between resistant and reference populations. Triphenyl phosphate (TPP), an inhibitor of esterases (ESTs), reduced the LC₅₀ value of abamctin in the populations as much as 1.73- to 3.73-fold. The activity of ESTs in these populations was also significantly different. Furthermore, inhibition of cytochrome P450 monooxygenases (CYP450s) by piperonyl butoxide (PBO) increased abamectin toxicity between 1.3- to 2.9-fold in tested populations. The highest ratios of synergism for DEM (5.86), TPP (3.73-fold), and PBO (2.91-fold) were observed in Shahre-e-Abrisham 1. It seems that GSTs and ESTs play a more important role in the resistance development against abamectin in the studied populations. High levels of resistance to abamectin in the collected populations from Isfahan Province shows the importance of insecticide resistance management based on the early detection of resistance and alternative use of insecticides.

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The present study was conducted to investigate the effects of two concentrations of Aloe vera gel (7.5 and 15%) on quantitative and qualitative characteristics of lime during storage time (20, 40, and 60 days). The study was performed as a factorial experiment in a completely randomized design with four replications. After dipping (5 minutes) and drying, the fruits were packaged in low-density polyethylene containers and transferred to cold storage at 4±1°C and 85±2% relative humidity. The results showed a decrease in fruit sourness and aroma and an increase in fruit bitterness during storage. The highest rate of chilling injury and decay belonged to the control. Aloe vera gel 15% had the lowest fruit bitterness (30%), decay, and malondialdehyde compared to the control on day 60 of storage. The Aloe vera gel enhanced peroxidase activity as an antioxidant enzyme and decreased defense-related enzymes such as phenylalanine ammonia-lyase activity. The maximum vitamin C was related to Aloe vera gel 15%. According to the results, Aloe vera gel, could not effectively control weight loss and firmness. During 60 days of storage, compared to the control, Aloe vera gel 15% increased Chroma index (2.07%) and vitamin C (26.37%), and prevented decay (100%), chilling injury (25.75%), bitterness (42.85%), and malondialdehyde (35.80%) of lime fruit.