scholarly journals Protease purification and characterization of a serine protease inhibitor from Egyptian varieties of soybean seeds and its efficacy against Spodoptera littoralis

2015 ◽  
Vol 55 (1) ◽  
pp. 16-25 ◽  
Author(s):  
Ashraf Oukasha Abd El-latif

Abstract Serine inhibitors have been described in many plant species and are universal throughout the plant kingdom. Trypsin inhibitors are the most common type. In the present study, trypsin and chymotrypsin inhibitory activity was detected in the seed flour extracts of four Egyptian varieties of soybean (Glycine max). The soybean variety, Giza 22, was found to have higher trypsin and chymotrypsin inhibitory potential compared to other tested soybean varieties. For this reason, Giza 22 was selected for further purification studies which used ammonium sulphate fractionation and DEAE-Sephadex A-25 column. Soybean purified proteins showed a single band on SDS-PAGE corresponding to a molecular mass of 17.9 kDa. The purified inhibitor was stable at temperatures below 60°C and was active at a wide range of pH, from 2 to 12 pH. The kinetic analysis revealed a non-competitive type of inhibition against trypsin and chymotrypsin enzymes. The inhibitor constant (Ki) values suggested that the inhibitor has higher affinity toward a trypsin enzyme than to a chymotrypsin enzyme. Purified inhibitor was found to have deep and negative effects on the mean larval weight, larval mortality, pupation, and mean pupal weight of Spodoptera littoralis. It may be concluded, that soybean protease inhibitor gene(s) could be potential targets for those future studies which are concerned with developing insect resistant transgenic plants

2013 ◽  
Vol 8 (10) ◽  
pp. 1934578X1300801 ◽  
Author(s):  
Arvind Dabhade ◽  
Priti Patel ◽  
Ulhas Patil

A thermo-stable, proteinaceous protease inhibitor (LPI) from Lawsonia inermis is reported. The LPI was purified from Lawsonia inermis seeds by subsequent ammonium sulfate precipitation, ion exchange chromatography (DEAE-Cellulose) and gel permeation chromatography (Sephadex-50). The purified protease inhibitor is effective against a wide range of proteases viz. papain, trypsin, pepsin and metallo-protease. The apparent molecular weight of the protease inhibitor is 19 kDa, determined by SDS-PAGE electrophoresis. The protease inhibitor was found to be stable at 70 °C for 30 min. It was also examined for antibacterial activity against Pseudomonas aeruginosa MTCC 7926 and Staphylococcus aureus NCIM 2079; the IC50 values of the purified LPI were 11.4 μg/mL and 16.6 μg/mL respectively.


2020 ◽  
Vol 89 (2) ◽  
pp. 101-112
Author(s):  
Soheir F. Abd El-Rahman ◽  
Sayeda S. Ahmed ◽  
Mahmoud H. Abdel Kader

Cotton leaf worm, Spodoptera littoralis (Boisduval, 1833), is one of the most dangerous pests in Egypt, causing economically significant losses of different crops. The present study was aimed to evaluate toxicological, biological and biochemical effects of two nanocomposites, Silver (Ag) and graphene oxide (GO) nanomaterials over magnesium chlorophillin (Mg-Chl/Ag and Mg-Chl/GO) at three concentrations (1, 10, 100 ml/L) against 2nd instar larvae of S. littoralis. The results showed that larval mortality rate was positively correlated with the increase of concentrations of the tested nanocomposites and time after exposure. The mortality rate in nanographene oxide composite were higher than nano silver. LC50 (lethal concentration of 50% of a group of test larvae) values were 10.27 and 16.14 mg/L at Mg-Chl/Ag and Mg-Chl/GO compound at 2 h exposure to light. Some biological aspects of S. littoralis resulted from the treated 2nd instars larvae with two nanocomposites were recorded. Larval, pupal durations, pupation, adult emergence rate and pupal weight were significantly lower in all concentrations compared to untreated larvae. All tested samples showed decrease in total carbohydrates, total proteins and total lipids. Obtained results suggest that using silver and graphene oxide nanomaterials over the magnesium chlorophillin would be a useful component for controlling S. littoralis.


2016 ◽  
Vol 56 (3) ◽  
pp. 279-290 ◽  
Author(s):  
Hala M. Mead ◽  
Samah N. El-Shafiey ◽  
Hend M. Sabry

Abstract The carried out investigations evaluated ovicidal activity of mahlab, Prunus mahaleb L. kernel oil against cotton leafworm, Spodoptera littoralis (Boisd.). The chemical constituents of the fixed oil of mahlab were analyzed using gas-liquid chromatography (GLC). Timnodonic (33.07%), oleic (28.71%) and linoleic (24.35%) were the basic fatty acids, while the major hydrocarbon and sterol were found to be heneicosane (62.57%) and β-sitosterol (10.57%). The LC50 values for the one-day-old egg masses were found to be more susceptible than 3-day-old ones. Moreover, the leaf dip technique occurred to be more efficient than spraying technique. The results also showed abnormalities in the external morphology of egg shell, chorion surface, shell imprints and aeropyles of S. littoralis eggs treated with mahlab and KZ oils as compared to a control by using scanning electron microscope. Generally, the tested oils significantly reduced the activities of transaminase enzymes (AST and ALT), acid and alkaline phosphatases and total soluble protein except mahlab oil on acid phosphatase as compared to a control. Additionally, the oils of both mahlab and KZ oil affected some biological aspects such as incubation period, larval duration, larval mortality and pupal weight comparing to a control.


2014 ◽  
Vol 6 (2) ◽  
pp. 633-637
Author(s):  
Sheetanshu Grover ◽  
Satvir Kaur Grewal ◽  
Anil Kumar Gupta ◽  
Gaurav Kumar Taggar

Helicoverpa armigera is one of the most devastating known pest causing major economic losses. Development of insect resistance against chemical pesticides and the environment concerns are forcing researchers to pave way towards biopesticides. Our study aimed to test the efficacy of seed flour from ten pigeon pea genotypes against H. armigera development. H. armigera fed with diet containing pigeon pea seed powder exhibited larval and pupal weight reduction and certain abnormalities. Larval weight reduction varied from 19 % (AL 1495) to 37 % (AL 1677). Increase in larval duration was seen in genotypes containing higher trypsin inhibitor, phenols and phytic acid. AL 1677 was found to be the most resistant genotype with 37 % larval weight reduction followed by AL 1735. Larval-pupal intermediates were observed in diets containing AL 1495 and AL 1747 seed powder. The pupal weight reduction varied from 15.5 % to 44.5 %. Results from this study suggest the inhibitory potential of PIs and other bioactive compounds such as phenols and phytic acid from pigeon pea cultivars against H. armigera development, that can be further exploited in pest management.


2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
Saswati Gayen ◽  
Uma Ghosh

Tannin acyl hydrolase produced extracellularly by the fungal strainPenicillium notatumNCIM 923 in mixed solid state fermentation of wheat bran and marigold flower in the ratio 4 : 1 was purified from the cell-free extract broth by ammonium sulphate fractionation followed by diethylaminoethyl-cellulose column chromatography. Tannase was purified by 19.89-fold with yield of 11.77%. The specific activity of crude tannase was found to be 1.31 U/mg protein while that of purified tannase was 22.48 U/mg protein. SDS-PAGE analysis indicated that the enzyme is dimeric with one major band of molecular mass 97 kDa and a very light band of molecular mass 43 kDa. Temperature of 35 to 40°C and pH 5 were optimum for tannase activity. The enzyme retained more than 60% of its stability at 60°C and 40% stability at pH 3 and 8, respectively.Kmwas found to be0.33×10-2 M andVmax=40 U/mg. Since the enzyme is active over a wide range of pH and temperature, it could find potential use in the food processing industry.


Author(s):  
Alexey Shcherbakov ◽  
Valentin Zhezmer

Department of hydraulic engineering and hydraulics FGBNU «VNIIGiM them. A.N. Kostyakova «has a long history. For many years, the department’s staff has been such scientists and water engineers with extensive experience as M.A. Volynov, V.S. Verbitsky, S.S. Medvedev, N.V. Lebedev, B.C. Panfilov, T.G. Voynich-Syanozhentsky, V.A. Golubkova, G.V. Lyapin and others. The department solved a wide range of tasks, the main areas of research were the following: – theoretical and applied hydrodynamics and hydraulics, with reference to the open channel flows that affect the state and level of safety of the hydraulic structures; – integrated use and protection of water bodies – water sources and water sources of water resources used in land reclamation; – development of measures and technical solutions for the protection of objects from the negative effects of water; – theoretical substantiation of works to improve the safety level of the GTS (declaration); – development and implementation of digitalization methods for solving design, construction, operation and control of landreclamation facilities. Currently, promising areas of research is the development of a decision-making algorithm in the designation of measures to rationalize the provision of resources to water amelioration. The algorithm is developed on the basis of a detailed study, systematization and processing of data both on safety and on the efficiency of systems and structures, ensuring the delivery of irrigation water of the required quality and in sufficient quantity from a water source to the field.


Author(s):  
M.A. Zemlianova ◽  
I.V. Tikhonova

Alumina refineries are among the leading sources of atmospheric air pollution with a wide range of pollutants hazardous to human respiratory organs. It is relevant to study and evaluate the occurrence of the risks for development of respiratory diseases in children living in the area affected by the emission components of an alumina refinery. We assessed air quality of the area under observation and comparison according to monitoring observations, risk of non-carcinogenic effects from the respiratory organs. The content of chemicals in the blood and urine adequate to risk factors was quantified. The structure of individual groups of respiratory diseases was analyzed. The causal relationships of violations of laboratory parameters with an increased content of chemicals in biological media were evaluated. It was found that an aerogenic exposure of chemical pollutants is formed on the territory with the production of metallurgical alumina. It determines the risk for development of respiratory diseases, exceeding an acceptable level up to 49.9 times. In the exposed children, the content of manganese, chromium, nickel, copper, xylenes, formaldehyde and aluminum, fluoride ion in the urine was increased to 4.2 times in relation to the indices in the comparison group. A high level of additional respiratory morbidity(1.8 times) was revealed. Chronic lymphoproliferative diseases of the nasopharynx and inflammatory diseases of the upper respiratory tract (up to 6.6 times more often), inflammatory diseases with a predominance of the mechanism of allergic inflammation ( up to 2.1 times more often)are more often detected in the framework of the respiratory diseases. Negative effects on the part of the respiratory system in the form of activation of antioxidant processes, the development of an inflammatory reaction, local, general and specific sensitization of the respiratory tract were established. It confirms the occurrence of the risks for the development of respiratory diseases in children in the exposure area of the chemical factors of alumina refinery-associated economic activity.


Pathogens ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 974
Author(s):  
Irina B. Ivshina ◽  
Maria S. Kuyukina ◽  
Anastasiia V. Krivoruchko ◽  
Elena A. Tyumina

Under conditions of increasing environmental pollution, true saprophytes are capable of changing their survival strategies and demonstrating certain pathogenicity factors. Actinobacteria of the genus Rhodococcus, typical soil and aquatic biotope inhabitants, are characterized by high ecological plasticity and a wide range of oxidized organic substrates, including hydrocarbons and their derivatives. Their cell adaptations, such as the ability of adhering and colonizing surfaces, a complex life cycle, formation of resting cells and capsule-like structures, diauxotrophy, and a rigid cell wall, developed against the negative effects of anthropogenic pollutants are discussed and the risks of possible pathogenization of free-living saprotrophic Rhodococcus species are proposed. Due to universal adaptation features, Rhodococcus species are among the candidates, if further anthropogenic pressure increases, to move into the group of potentially pathogenic organisms with “unprofessional” parasitism, and to join an expanding list of infectious agents as facultative or occasional parasites.


Author(s):  
Eman Mohammed Abd-ElAzeem ◽  
Warda Ahmed Zaki El-Medany ◽  
Hend Mohammed Sabry

AbstractBiological activities of spores and metabolites of some fungi isolated from dead larva of the spiny bollworms (SBW), Earias insulana (Boisd.) (Lepidoptera: Noctuidae), against the newly hatched larvae of the pest were carried out. Results showed that the fungi Metarhizium anisopliae, Acremonium sp., and Paecilomyces variotii had affected the newly hatched larvae of (SBW). Acremonium sp. was the most potent one as it had the highest newly hatched larval mortality percentage (65 and 58.33%) for its spore suspension and metabolites, respectively, while the lowest one (41%) was for P. variotii metabolites. Also, spore suspensions of the all fungal isolates had the highest larval mortality than fungal metabolites. Studying the enzymatic activity showed that Acremonium sp. produced protease enzyme on media containing gelatin, which caused the highest larval mortality (72.22%).These isolates showed different effects on all stages of the pest and decreased pupal weight, adult emergence percentages, deposited eggs, and hatchability percentages than the control. Identification of Acremonium sp. EZ1 was confirmed using 18 s rRNA and its accession number MN25101.


2005 ◽  
Vol 387 (1) ◽  
pp. 271-280 ◽  
Author(s):  
Seonghun KIM ◽  
Sun Bok LEE

The extremely thermoacidophilic archaeon Sulfolobus solfataricus utilizes D-glucose as a sole carbon and energy source through the non-phosphorylated Entner–Doudoroff pathway. It has been suggested that this micro-organism metabolizes D-gluconate, the oxidized form of D-glucose, to pyruvate and D-glyceraldehyde by using two unique enzymes, D-gluconate dehydratase and 2-keto-3-deoxy-D-gluconate aldolase. In the present study, we report the purification and characterization of D-gluconate dehydratase from S. solfataricus, which catalyses the conversion of D-gluconate into 2-keto-3-deoxy-D-gluconate. D-Gluconate dehydratase was purified 400-fold from extracts of S. solfataricus by ammonium sulphate fractionation and chromatography on DEAE-Sepharose, Q-Sepharose, phenyl-Sepharose and Mono Q. The native protein showed a molecular mass of 350 kDa by gel filtration, whereas SDS/PAGE analysis provided a molecular mass of 44 kDa, indicating that D-gluconate dehydratase is an octameric protein. The enzyme showed maximal activity at temperatures between 80 and 90 °C and pH values between 6.5 and 7.5, and a half-life of 40 min at 100 °C. Bivalent metal ions such as Co2+, Mg2+, Mn2+ and Ni2+ activated, whereas EDTA inhibited the enzyme. A metal analysis of the purified protein revealed the presence of one Co2+ ion per enzyme monomer. Of the 22 aldonic acids tested, only D-gluconate served as a substrate, with Km=0.45 mM and Vmax=0.15 unit/mg of enzyme. From N-terminal sequences of the purified enzyme, it was found that the gene product of SSO3198 in the S. solfataricus genome database corresponded to D-gluconate dehydratase (gnaD). We also found that the D-gluconate dehydratase of S. solfataricus is a phosphoprotein and that its catalytic activity is regulated by a phosphorylation–dephosphorylation mechanism. This is the first report on biochemical and genetic characterization of D-gluconate dehydratase involved in the non-phosphorylated Entner–Doudoroff pathway.


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