The use of entomopathogenic fungi (EF) in recent years has been highly effective against the different orders of insects considered pests of agricultural importance and their conidia have been commonly applied, but it has been reported that these are sensitive to the environmental conditions. For this reason, biopesticides products have been formulated based on secondary metabolites, recently. These biomolecules participate as biological control agent, such as: cyclic depsipeptides, amino acids, polyketides, polyphenols and terpenoids, affecting their morphology, life cycle and insect behavior. The use of secondary metabolites of entomopathogenic fungi opens the possibility of application in a more efficient way for the control of agricultural pests in a compatible with the environment and human health; therefore, it is important to know, analyzing the type of molecules, their effects, and their different methods of application.
Abstract Remote control enzyme technology is widely used today through resonance. In this study, we showed that the use of frequencies of the catalase enzyme itself to increase enzymatic rate is successful not only in test tubes but also remotely. The present study also suggests that, under optimal temperature, the use of bovine catalase frequency (the specific frequency of that enzyme) has a superior rate promoting vibration than the human catalase frequency, and so increases very significantly the chemical rate of bovine catalase (about 120% at 40ºC). It also suggests that bovine catalase subjected to bovine and human frequencies with catalase bound NADP+ experienced more resonance weight towards NADP+ and so were more slowly reduced back to catalase bound NADPH, increasing compound II formation rate, and slowing down the catalase activity rate.
Plant essential oils and their constituents have proven to be very effective against insects, especially termites. They are the best alternative to synthetic pesticides that are harmless to the environment and human health. In the present study, different enzymes, namely alkaline phosphatase, acid phosphatase, glutamate-oxaloacetate transaminase, glutamate-pyruvate transaminase and acetylcholinesterase, were evaluated in Indian termite Odontotermes obesus in a combination preparation based on Tagetes erecta essential oil. For the study of anti-termite effects against worker termites were treated topically with 40% and 80% of the 24-hour LD50 values of various combination formulations. Subsequent bioassays at 40% and 80% of LD50 of combinatorial preparations S-AST-A, S-AST-B, and S-AST-C resulted in a significant (p > 0.05) decrease in glutamate-oxaloacetate transaminase. After 16 hours of treatment, they were 87.92%, 80.17%, 89.30%, 79.17%, 81.92% and 73.17% respectively. In vivo exposures of 40% and 80% of the LD50 of the combination formulation caused a highly significant (p > 0.05) reduction (p > 0.05) of all test enzymes tested compared to controls. The effects of different oil formulations exhibited time- and dose-dependent responses, resulting in physiological changes in the concentrations of various enzymes. The combined mixture of Tagetes erecta essential oils has significantly better anti-termite ability compared to inorganic insecticides. Findings from this study will help support termite control in fields, gardens and homes in a sustainable way, without the downsides of insecticide resistance and pollution. These could potentially be used to produce commercial formulations for use against pests.
Wheat arabinoxylan (water-insoluble fraction) contains ~36% arabinose which may include both singly or doubly substitutions at C2/C3 of the Xylp units. α-L-Arabinofuranosidses (ABFs) of two GH families were analyzed for their respective activities on the hydrolysis of Xylp-Araf. BaABF (GH43) produced twice the yield of arabinose residues from the heteroxylan compared to AnABF (GH51) under the same reaction conditions. Two endo-xylanases (of GH10 and 11) also showed differential hydrolytic activities on the Xylp chain, with the GH10 XYN-ATM double the amount of reducing sugar yield (as xylose equivalent) than using the GH11 XYN-M3. When the ABF and XYN were combined in optimial ratios, a synergistic increase of 73.8% in arabinose yield was observed.
In the present, investigation effects of sub-lethal dose of purified paper wasp Ropalidia marginata venom toxins were evaluated on important metabolic enzymes i.e.ALP ACP, GPT, GOT, LDH, and AchE enzyme activity in serum, liver, and gastrocnemius muscles of albino mice.Alkaline phosphatase was found to be increased up to 119.9% at the 6 th hr of the toxin injection in comparison to control.This elevation may be due to cytolysis.Maximum increase i.e., 153.33% level of glutamate pyruvate transaminase (GPT) was found at 6 hrs of 40% of 24-h LD 50 treatment while it was found to be 151.1% at 6 hrs of 24 hr 80% of LD 50 , venom injection.A significant elevation was observed in LDH activity in serum, liver, and muscles, while the activity of AchE was decreased in serum, liver, and gastrocnemius muscles of albino mice after injecting the sub-lethal dose of Ropalidia marginata venom.This increase in the activity of LDH produces liver damage, massive disintegration and necrosis of hepatic cells.This elevation in LDH level led to a significant increase in the glucose catabolism and elevated oxidative stress in muscle and liver cells.It also displays insufficient oxygen supply and consequently leads to cell death.In experimental animals, venom toxin treatment decreased AchE level, and animals showed muscular paralysis.When mice were treated with 40% and 80% of 24-h LD 50 of purified venom caused a significant (p < 0.05) elevation in the level of ACP, GOT, GPT, and LDH while the reduction in ALP and AChE level.Present study will be useful in the development of prototypes for study of pharmacological and therapeutic effects of various venom toxins.For this purpose structure activity relationship of enzyme and venom toxin, its due interaction to various metabolic enzymes and receptors must be explored.
Combinatorial enzyme technology was applied for the conversion of wheat insoluble arabinoxylan to oligosaccharide structural variants. The digestive products were fractionated by Bio-Gel P4 column and screened for bioactivity. One fraction pool was observed to exhibit antimicrobial property resulting in the suppression of cell growth of the test organism ATCC 8739 E. coli. It has a MIC value of 1.5% (w/v, 35°C, 20 hr) and could be useful as a new source of prebiotics or preservatives. The present results further confirm the science and useful application of combinatorial enzyme approach.
In the present investigation, various bioassays were conducted to evaluate the alteration in levels of various enzymes i.e. alkaline phosphatase, acid phosphatase, glutamate oxaloacetate transaminase, glutamate pyruvate transaminase and acetylcholinesterase.For this purpose worker termites were treated with sub-lethal doses of 40% and 80% of 24 hrs LD 50 of latex-based combinatorial formulations and observations were taken at 4 hours intervals up to 24 hours.Ficus benghalensis crude latex and its combinatorial mixtures, like C-MLT-A, CU-MLT-A, AQ-MLT, P-MLT and EA-MLT significantly altered the level of enzymes in Odontotermes obesus and this effect found time and dose-dependent.Reduction or increase in enzymes was calculated by using the corresponding control.The maximum decrease in acid phosphatase level was observed at 16 h when termites were treated with 80% of LD 50 of B-MLT-B and P-MLT i.e. 82.84% at 16 h of treatment.A similar dose caused a very slight decrease in glutamate pyruvate transaminase at 4 h of treatment but with the increase in time.Further decrease was noted in other successive treatments that were significant at p > 0.05.Similarly, 40% and 80% of LD 50 of the C-MLT-B mixture caused significant (p > 0.05) decrease in alkaline phosphatase and acid phosphatase level at 16 h treatment i.e. 93.42%, 89.46% and 95.89%, 88.17% respectively.The level of acetylcholinesterase was also found to be decreased when termites were treated with 40% and 80% of LD 50 of C-MLT-B mixture i.e. 92.72% and 97.27% respectively.All the above alterations noted in levels of various enzymes confirm the action of latex ingredients on worker termites that were anti-feedant or deterrent types.These ingredients can be used to control not only termites but also other phytophagous insects.
In the present investigation, in vivo effects of purified ticks’ saliva toxin were evaluated on the level of certain important cellular metabolic enzymes i.e. acid phosphatase (ACP), alkaline phosphatase (ALP), glutamate pyruvate transaminase, glutamate oxaloacetate transaminase and lactic dehydrogenase. For this purpose, sub-lethal doses, 40% and 80% of 24 h LD50 purified saliva toxins of Rhipicephalus microplus (Canestrini, 1888) were injected subcutaneously in the albino mice. In treated mice saliva toxins targeted membrane-bound enzymes i.e. serum acid phosphatase and alkaline phosphatase, its level was increased from 118.30% to 163.63% at the 6th hr in comparison to the control. Besides this, the levels of serum glutamate pyruvate transaminase (GPT) and glutamate oxaloacetate transaminase (GOT) and lactic dehydrogenase (LDH) also increased up to 161.11% (at 6th hr), 148.27 (at 8th hr) and 125.45% (at 6th hr) respectively in comparison to control. An increase in the level of LDH showed insufficient oxygen supply, massive disintegration of cells and leakage of the enzyme into the circulation. It clearly indicated the toxic effects of saliva toxins on the membrane of blood cells, hepatocytes and myocardial muscle cell functions in albino mice. On the other hand activity of acetyl cholinesterase was reduced by 65.51% at the 6th hr of the saliva toxin injection in comparison to the control. This inhibition of acetyl cholinesterase activity caused the accumulation of acetylcholine molecules at the synaptic junctions and led to prolonged activation of acetylcholine receptors. It caused permanent stimulation of nerves and muscle cells that may result in muscular paralysis and finally death of the animal.
In the present study, Citrus maxima essential crude oil extract was used to prepare combinatorial formulations and workers of Indian white termite Odontotermes obesus were treated topically with 40% and 80% of 24 hr LD50 values of these formulations. In subsequent bioassays levels of various enzymes i.e. alkaline phosphatase, acid phosphatase, glutamate oxaloacetate transaminase, glutamate pyruvate transaminase and acetylcholinesterase were evaluated to determine the anti-termite efficacy of Citrus maxima essential oil based combinatorial formulations. S-RET-A, S-RET-B and S-RET-C caused significant (p > 0.05) decrease in glutamate oxaloacetate transaminase i.e. 87.47%, 86.81% and 81.77% & 82.04%, 79.39% and 74.75% respectively at 16 h treatment. In vivo exposure of 40% and 80% of LD50 of combinatorial formulations caused very significant (p > 0.05) reduction in all the test enzymes i.e. alkaline phosphatase, acid phosphatase, glutamate oxaloacetate transaminase and glutamate pyruvate transaminase expect acetylcholinesterase levels after 16 h all tested treatments in comparison to control. Both dose-response and time period were found important in physiological alteration in levels of various enzymes. Combinatorial mixtures of Citrus essential oils have shown synergistic activity against termites. The research findings of the present study would help termite control in crop fields, gardens and houses in a sustainable way.
The use of enzymes is replacing chemicals in many industrial production processes because of the eco-friendly nature of enzymes which do not generate greenhouse gases and have reduced the demand for energy in industries. To meet the ever-increasing demand for enzymes in many industries and survive the harsh production conditions, microbial sources of enzyme production are the most preferred source for industrial enzyme production because the microbes are readily available, they grow at a very fast rate, and they can be genetically manipulated to produce enzymes which can perform optimally at different industrial production conditions. Microbial enzymes have found so many applications in various industries (textiles, leather, paper and pulp, research and development, pharmaceutical, agriculture, detergent, waste, biorefineries, photography and food industries), thus making them very essential in several industrial production processes. Here in this review, the application of some important microbial enzymes in food industry and the microbial sources for the enzymes are discussed.
Hormone Receptor positive (HR+) breast cancer is the most common malignancy in women. New strategies in the treatments have targeted the estrogen biosynthesis pathways including the inhibition of the aromatase and 17β-HSD1 enzymes. The present work, describes the study of a new family of 9 hybrid compounds derived from estrone attached to a coumarin fragment, linked through different lengths of hydrocarbon chains. The activity of these compounds was evaluated by molecular docking with two relevant enzymes in breast cancer (HR+). It has been proposed nine compounds as 17β-HSD1 inhibitors and six as aromatase inhibitors. We found important interactions with key amino acids at the orthosteric site of each enzyme and their score values compared to the crystallographic ligand. The in silico analysis showed good score values in the proposed compounds, where the steroidal portion presented important interactions with Met374 and Tyr155 in aromatase and in 17β-HSD1 respectively. Highlighting Compounds 2, 5 and 8 with an aromatic ring at the C4 position of the coumarin moiety, which favored arene-H type interactions essential for protein-ligand recognition. In addition, the results related to the 17β-HSD1 enzyme demonstrated how the length of the linker influences the interaction; the best score was found for derivative 8 with a chain of 8 methylenes.
Lipase producing Aspergillus was isolated from soil collected from a refuse dump site located at Awka, Anambra State, Nigeria using standard microbiology and biochemical techniques.Crude extract of lipase was produced after a successful screening of the isolates using mineral broth containing p-NPP through submerged fermentation system with optimized physiologic conditions.Three steps of purification were carried out: Ammonium sulphate, dialysis and gel filtration (sephadex G-150).Crude extract was precipitated by using 70% saturation of ammonium sulphate at pH 6.0 which gave the optimum precipitation of the protein with specific activity of 260.56 U/mg.Precipitation using ammonium sulphate carried out at pH 6.5 and 8.0 gave specific activity of 217 U/mg of the protein.The precipitates were further desalted through dialysis for twelve hours and specific activity of 343.20 U/mg was recorded from the dialysate afterwards.Further purification was done by using sephadex G-150 and specific activity of 490.55 U/mg was recorded from the active pooled fractions.The purification table showed a 2.32 purification folds of lipase was gotten after gel filtration (sephadex G-150) with a lipase percentage yield of 2.00%.The specific activity of lipase increased from 211.81 to 490.55 U/mg.Characterization of β-galactosidase gave optima pH and temperature of the enzyme at 6.0 and 60˚C respectively.Kinetic constants: K m and V max values were obtained at various concentrations of p-NPP where 0.32 mM and V max of 200.00 μmol/min respectively.Ca 2+ and Co 2+ showed greater effect on lipase activity in a concentration-dependent manner (0.03 -0.05 M) when compared to Mn 2+ and Fe 2+ .The results from this study have shown that lipase produced from filamentous Aspergillus has a wide range of activity over physiologic conditions in regards to industrial and clinical standard operational procedures.
An α-L-arabinofuranosidase (ARF) gene of 1503 bp was synthesized, subcloned into pET26b vector, and expressed in Escherichia coli.The enzyme was purified in active form, and consisted of 500 amino acid residues, corresponding to 55 kD based on SDS-PAGE.The affinity-purified protein was characterized using arabinofuranosyl xylooligosaccharides (AXOS) as substrates.The pH effect was investigated showing an optimum at pH 5.5.XaARF catalyzed the cleavage of arabinose at C3 of the xylopyranosyl unit efficiently if the arabinofuranosyl substitution was at the terminal compared to internal xylose units.The enzyme was able to act on di-substituted xylopyranosyl units with the first cleavage at C3 followed by C2 linkages.
Biochar offers several benefits as a soil amendment, including increased soil fertility, carbon sequestration, and water-holding capacity in nutrient-poor soils.Here, we performed a series of enzyme assays on pine biochar-amended soils, comparing multiple enzyme activities (EAs) simultaneously determined in the same soil sample vs. the sum of individual EAs involved in the C, N, S, and P cycles to provide information of the impacts of biochar on biogeochemical cycling.The combination of these four EAs has been considered an indicator of soil health due to their role in the reactions that release bioavailable nutrients in the cycling of C (β-glucosidase), N and C (β-glucosaminidase), P (acid phosphomonoesterase), and S (arylsulfatase) in soils.Comparisons of the theoretical EAs and the CNPS activity assay approaches in the biochar-modified soil revealed similar activity trends with the different concentrations of added biochar.Two years after adding biochar, study results showed the amended soils did not retain more pNP substrate than the un-amended control soils in three different pH buffers (5.5, 5.8, and 6.5) commonly used in EA reactions.Finally, we performed a third experiment to determine if the biochar previously added to the EAs interfered with the reactions' enzyme or substrate.The results indicated that greater activity was measured using the combined assay, which suggests the CNPS activity method was less affected by biochar than the individual EAs.Our findings indicate that the potential soil biochemical-health index, CNPS activity (combination of four enzymes) assay is more robust than the individual EAs and can be used as an alternative tool to monitor soil functioning.
Thermostable α-amylases hold a very important place in commercial industrial applications in Sri Lanka.Therefore, the main aim of this study was to identify superior Bacillus strain and optimize growth conditions that could yield high α-amylase production.Three Bacillus strains, B. amyloliquefaciens ATCC 23350, B. licheniformis ATCC 14580 and B. megaterium ATCC 14581 were used for the study.Shake flask culture experiments were conducted to identify the effect of various fermentation conditions such as growth temperature, incubation period, carbon source, nitrogen source, initial pH and carbon concentration on extracellular α-amylase production.DNSA assay was carried out to determine the enzyme activity.The highest temperature for enzyme activity was reported by B. licheniformis at 85˚C, followed by B. amyloliquefaciens at 75˚C and B. megaterium at 45˚C.Both B. amyloliquefaciens and B. licheniformis were able to give their optimum enzyme production at 37˚C, while B. megaterium at 30˚C in 150 rpm with initial pH of 7. B. licheniformis and B. amyloliquefaciens gave their optimum yield of the enzyme after 48 h of incubation while B. megaterium gave after 24 h of incubation.Among the carbon sources tested cassava starch was able to give the highest enzyme production.For B. amyloliquefaciens, the highest yield of the enzyme was obtained with 2% of starch, tryptone as a nitrogen source and initial pH of 7. Maximum enzyme production for B. licheniformis was obtained with 1.5% of starch, KNO 3 as a nitrogen source and initial pH of 6.For B. megaterium 1% of starch, tryptone and pH 7.5 induced the optimum α-amylase production.According to the results obtained, B. amyloliquefaciens is the highest thermostable alpha amylase producer.However, according to the industrial requirement, B. licheniformis can also be used as an enzyme producer due to its stability in higher temperatures.
