Search results for: enzymatic inhibition activity

Authors: Boutemak Khalida, Dahmani Siham

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Garden cress (Lepidium Sativum L.) belonging to the family Brassicaceae, is edible growing annual herb. Its various parts (roots, leaves and seeds) have been used to treat various human ailments. Its seed extracts have been screened for various biological activities like hypotensive, antimicrobial, bronchodilator, hypoglycaemic and antianemic. The aim of the present study is to optimize the process parameters (cellulase concentration and incubation time) of enzymatic pre-treatment of the garden cress seeds and to evaluate the effect of cellulase pre-treatment of the crushed seeds on the oil yield, physico-chemical properties and antibacterial activity and comparing to non-enzymatic method. The optimum parameters of cellulase pre-treatment were as follows: cellulase of 0,1% w/w and incubation time of 2h. After enzymatic pre-treatment, the oil was extracted by n-hexane for 1.5 h, the oil yield was 4,01% for cellulase pre-treatment as against 10,99% in the control sample. The decrease in yield might be caused a result of mucilage. Garden cress seeds are covered with a layer of mucilage which gels on contact with water. At the same time, the antibacterial activity was carried out using agar diffusion method against 4 food-borne pathogens (Escherichia coli, Salmonella typhi,Staphylococcus aureus, Bacillus subtilis). The results showed that bacterial strains are very sensitive to the oil with cellulase pre-treatment. Staphylococcus aureus is extremely sensitive with the largest zone of inhibition (40 mm), Escherichia coli and salmonella typhi had a very sensitive to the oil with a zone of inhibition (26 mm). Bacillus subtilizes is averagely sensitive which gave an inhibition of 16 mm. But it does not exhibit sensivity to the oil without enzymatic pre-treatment with a zone inhibition (< 8 mm). Enzymatic pre-treatment could be useful for antimicrobial activity of the oil, and hold a good potential for use in food and pharmaceutical industries.

Keywords: Lepidium sativum L., cellulase, enzymatic pretreatment, antibacterial activity.

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Authors: Rafayel Azizyan, Valeri Arakelyan, Aram Gevorgyan, Varduhi Balayan, Emil Gevorgyan

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Currently, mathematical and computer modeling are widely used in different biological studies to predict or assess behavior of such complex systems as biological ones. This study deals with mathematical and computer modeling of bi-substrate enzymatic reactions, which play an important role in different biochemical pathways. The main objective of this study is to represent the results from in silico investigation of bi-substrate enzymatic reactions in the presence of uncompetitive inhibitors, as well as to describe in details the inhibition effects. Four models of uncompetitive inhibition were designed using different software packages. Particularly, uncompetitive inhibitor to the first [ES1] and the second ([ES1S2]; [FS2]) enzyme-substrate complexes have been studied. The simulation, using the same kinetic parameters for all models allowed investigating the behavior of reactions as well as determined some interesting aspects concerning influence of different cases of uncompetitive inhibition. Besides that shown, that uncompetitive inhibitors exhibit specific selectivity depending on mechanism of bi-substrate enzymatic reaction.

Keywords: mathematical modeling, bi-substrate enzymatic reactions, reversible inhibition

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Authors: J. B. Minari, O. B. Oloyede, A. A. Odutuga

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Myeloperoxidase is the most abundant enzyme found in the polymorphonuclear neutrophil and is known to play a central role in the host defense system of the leukocyte. The enzyme has been reported to interact with some drugs to generate free radical which inhibits its activity. This study investigated the effects of artesunate on the activity of the enzyme and the subsequent effect on the host immune system. In investigating the effects of the drugs on myeloperoxidase, the influence of concentration, pH, partition ratio estimation and kinetics of inhibition were studied. This study showed that artesunate is concentration-dependent inhibitor of myeloperoxidase with an IC50 of 0.078mM. Partition ratio estimation showed that 60 enzymatic turnover cycles are required for complete inhibition of myeloperoxidase in the presence of artesunate. The influence of pH on the effect of artesunate on the enzyme showed least activity of myeloperoxidase at physiological pH. The kinetic inhibition studies showed that artesunate caused a competitive inhibition with an increase in the Km value from 0.12mM to 0.26mM and no effect on the Vmax value. The Ki value was estimated to be 2.5mM. The results obtained from this study show that artesunate is a potent inhibitor of myeloperoxidase and it is capable of inactivating the enzyme. It is considered that the inhibition of myeloperoxidase in the presence of artesunate as revealed in this study may partly explain the impairment of polymorphonuclear neutrophil and consequent reduction of the strength of the host defense system against secondary infections.

Keywords: myeloperoxidase, artesunate, inhibition, nuetrophill

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Authors: Delgado-Meza M., Minor-Pérez H.

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Protease is the generic name of enzymes that hydrolyze proteins. These are classified in the subgroup EC3.4.11-99X of the classification enzymes. In food technology the proteolysis is used to modify functional and nutritional properties of food, and in some cases this proteolysis may cause food spoilage. In general, seafood and rainbow trout have accelerated decomposition process once it has done its capture, due to various factors such as the endogenous enzymatic activity that can result in loss of structure, shape and firmness, besides the release of amino acid precursors of biogenic amines. Some studies suggest the use of protease inhibitors from legume as biological regulators of proteolytic activity. The enzyme inhibitors are any substance that reduces the rate of a reaction catalyzed by an enzyme. The objective of this study was to evaluate the reduction of the proteolytic activity of enzymes in extracts of rainbow trout with protease inhibitors obtained from chickpea flour. Different proportions of rainbow trout enzyme extract (75%, 50% and 25%) and extract chickpea enzyme inhibitors were evaluated. Chickpea inhibitors were obtained by mixing 5 g of flour in 30 mL of pH 7.0 phosphate buffer. The sample was centrifuged at 8000 rpm for 10 min. The supernatant was stored at -15°C. Likewise, 20 g of rainbow trout were ground in 20 mL of phosphate buffer solution at pH 7.0 and the mixture was centrifuged at 5000 rpm for 20 min. The supernatant was used for the study. In each treatment was determined the specific enzymatic activity with the technique of Kunitz, using hemoglobin as substrate for the enzymes acid fraction and casein for basic enzymes. Also biuret protein was quantified for each treatment. The results showed for fraction of basic enzymes in the treatments evaluated, that were inhibition of endogenous enzymatic activity. Inhibition values compared to control were 51.05%, 56.59% and 59.29% when the proportions of endogenous enzymes extract rainbow trout were 75%, 50% and 25% and the remaining volume used was extract with inhibitors. Treatments with acid enzymes showed no reduction in enzyme activity. In conclusion chickpea flour reduced the endogenous enzymatic activity of rainbow trout, which may favor its application to increase the half-life of this food. The authors acknowledge the funding provided by the CONACYT for the project 131998.

Keywords: rainbouw trout, enzyme inhibitors, proteolysis, enzyme activity

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Authors: Khalida Boutemak, Nasssima Benali, Nadji Moulai-Mostefa

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Effect of enzyme on the yield and chemical composition of Artemisia campestris essential oil is reported in the present study. It was demonstrated that enzyme facilitated the extraction of essential oil with increase in oil yield and did not affect any noticeable change in flavour profile of the volatile oil. Essential oil was tested for antibacterial activity using Escherichia coli; which was extremely sensitive against control with the largest inhibition (29mm), whereas Staphylococcus aureus was the most sensitive against essential oil obtained from enzymatic pre-treatment with the largest inhibition zone (25mm). The antioxidant activity of the essential oil with hemicellulase pre-treatment (EO2) and control sample (EO1) was determined through reducing power. It was significantly lower than the standard drug (vitamin C) in this order: vitamin C˃EO2˃EO1.

Keywords: Artemisia campestris, enzyme pre-treatment, hemicellulase, antibacterial activity, antioxidant activity

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Authors: Catherine Joke Adeseko

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Enzymatic browning is an economically important disorder that degrades organoleptic properties and prevent the consumer from purchasing fresh fruit and vegetables. Prevention and control of enzymatic browning in fruit and its product is imperative. Therefore, this study sought to investigate the catalytic effect of polyphenol oxidase (PPO) in the adverse browning of African bush mango (Irvingia gabonensis) fruit peel and pulp. PPO was isolated and purified, and its physicochemical properties, such as the effect of pH with SDS, temperature, and thermodynamic studies, which invariably led to thermal inactivation of purified PPO at 80 °C, were evaluated. The pH and temperature optima of PPO were found at 7.0 and 50, respectively. There was a gradual increase in the activity of PPO as the pH increases. However, the enzyme exhibited a higher activity at neutral pH 7.0, while enzymatic inhibition was observed at acidic region, pH 2.0. The presence of SDS at pH 5.0 downward was found to inhibit the activity of PPO from the peel and pulp of I. gabonensis. The average value of enthalpy (ΔH), entropy (ΔS), and Gibbs free energy (ΔG) obtained at 20 min of incubation and temperature 30 – 80 °C were respectively 39.93 kJ.mol-1, 431.57 J.mol-1 .K-1 and -107.99 kJ.mol-1 for peel PPO, and 37.92 kJ.mol-1, -442.51J.mol-1.K-1, and -107.22 kJ.mol-1 for pulp PPO. Thermal inactivation of PPO from I. gabonensis exhibited a reduction in catalytic activity as the temperature and duration of heat inactivation increases using catechol, reflected by an increment in k value. The half-life of PPO (t1/2) decreases as the incubation temperature increases due to the instability of the enzyme at high temperatures and was higher in pulp than peel. Both D and Z values decrease with increase in temperature. The information from this study suggests processing parameters for controlling PPO in the potential industrial application of I. gabonensis fruit in order to prolong the shelf-life of this fruit for maximum utilization.

Keywords: enzymatic, browning, characterization, activity

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Authors: Asma Meddour, Mouloud Yahia, Afaf Benhouda, Souhila Benbia, Hachani Khadhraoui

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Peripheral analgesic activity of defatted methanol extract of root barks of Capparis spinosa was tested orally at the dose of 100 and 200 mg/kg against pain induced by acetic acid in rats. The dose of 200 mg/kg presents significant analgesic effect with a percentage of inhibition of torsions of 88.51% compared to the positive control which is the acetylsalicylic acid which represents a percentage of inhibition of 92.55%. The dose of 100 mg/kg presents a percentage of inhibition of 81.68%.

Keywords: peripheral analgesic activity, Capparis spinosa, percentage of inhibition of torsions, chemical sciences

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Authors: Rajesh A. Rane, Shital S. Naphade, Rajshekhar Karpoormath

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Thiozolidin-4-one, a mimic of thiazolobenzimidazole (TBZ) has drawn many attentions due to its potent and selective inhibition against the HIV-1 and low toxicity by binding to the allosteric site of the reverse transcriptase (RT) as a non-nucleoside RT inhibitor (NNRTI). Similarly, marine bromopyrrole alkaloids are well known for their diverse array of anti-infective properties. Hence, we have reported synthesis and in vitro HIV-1 RT inhibitory activity of a series of 4-thiazolidinone-bromopyrrole alkaloid hybrids tethered with amide linker. The results of in vitro HIV-1 RT kit assay showed that some of the compounds, such as 4c, 4d, and 4i could effectively inhibit RT activity. Among them, compounds 4c having 4-chlorophenyl substituted 4-thiazolidione ring was the best one with the IC50 value of 0.26 µM. The sturdy emerges with key structure-activity relationship that pyrrole-NH-free core benefited inhibition against HIV-1 RT inhibition. This study identified conjugate 4c with potent activity and selectivity as promising compound for further drug development to HIV.

Keywords: antiviral drugs, bromopyrrole alkaloids, HIV-1 RT inhibition, 4-thiazolidinone

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Authors: Gulnur Arabaci

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Heavy metals are one of the major groups of contaminants in the environment and many of them are toxic even at very low concentration in plants and animals. However, some metals play important roles in the biological function of many enzymes in living organisms. Metals such as zinc, iron, and cooper are important for survival and activity of enzymes in plants, however heavy metals can inhibit enzyme which is responsible for defense system of plants. Polyphenol oxidase (PPO) is a copper-containing metalloenzyme which is responsible for enzymatic browning reaction of plants. Enzymatic browning is a major problem for the handling of vegetables and fruits in food industry. It can be increased and effected with many different futures such as metals in the nature and ground. In the present work, PPO was isolated and characterized from green leaves of red poppy plant (Papaver rhoeas). Then, the effect of some known antibrowning agents which can form complexes with metals and metals were investigated on the red poppy PPO activity. The results showed that glutathione was the most potent inhibitory effect on PPO activity. Cu(II) and Fe(II) metals increased the enzyme activities however, Sn(II) had the maximum inhibitory effect and Zn(II) and Pb(II) had no significant effect on the enzyme activity. In order to reduce the effect of heavy metals, the effects of metal-antibrowning agent complexes on the PPO activity were determined. EDTA and metal complexes had no significant effect on the enzyme. L-ascorbic acid and metal complexes decreased but L-ascorbic acid-Cu(II)-complex had no effect. Glutathione–metal complexes had the best inhibitory effect on Red poppy leaf PPO activity.

Keywords: inhibition, metal, red poppy, poly phenol oxidase (PPO)

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Authors: Rajesh Kumar, Anil Kamboj

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Inhibition of α- amylase play a vital role in the clinical management of postprandial hyperglycemia. Although, powerful synthetic inhibitors are available, natural inhibitors are potentially safer. The present study was carried out to evaluate α- amylase inhibition activity from hydroalcoholic extracts from aerial parts of Cestrum nocturnum. Hydroalcoholic extract was prepared by Soxhletation Method. The extract showed strong inhibition towards α- amylase activity and IC50 value were 45.9 µg. This In vitro studies indicate the potential of C. nocturnum in the development of effective anti-diabetic agents.

Keywords: α- amylase, cestrum nocturnum, hyperglycemia, hydroalcoholic extracts, diabetes

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Authors: Anshu Siwach, Qianlai Zhuang, Ratul Baishya

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Context: This study focuses on the influence of moss cover and seasonality on soil microbial biomass and enzymatic activity in different Central Himalayan temperate forest types. Soil microbial biomass and enzymes are key indicators of microbial communities in soil and provide information on soil properties, microbial status, and organic matter dynamics. The activity of microorganisms in the soil varies depending on the vegetation type and environmental conditions. Therefore, this study aims to assess the effects of moss cover, seasons, and different forest types on soil microbial biomass carbon (SMBC), soil microbial biomass nitrogen (SMBN), and soil enzymatic activity in the Central Himalayas, Uttarakhand, India. Research Aim: The aim of this study is to evaluate the levels of SMBC, SMBN, and soil enzymatic activity in different temperate forest types under the influence of two ground covers (soil with and without moss cover) during the rainy and winter seasons. Question Addressed: This study addresses the following questions: 1. How does the presence of moss cover and seasonality affect soil microbial biomass and enzymatic activity? 2. What is the influence of different forest types on SMBC, SMBN, and enzymatic activity? Methodology: Soil samples were collected from different forest types during the rainy and winter seasons. The study utilizes the chloroform-fumigation extraction method to determine SMBC and SMBN. Standard methodologies are followed to measure enzymatic activities, including dehydrogenase, acid phosphatase, aryl sulfatase, β-glucosidase, phenol oxidase, and urease. Findings: The study reveals significant variations in SMBC, SMBN, and enzymatic activity under different ground covers, within the rainy and winter seasons, and among the forest types. Moss cover positively influences SMBC and enzymatic activity during the rainy season, while soil without moss cover shows higher values during the winter season. Quercus-dominated forests, as well as Cupressus torulosa forests, exhibit higher levels of SMBC and enzymatic activity, while Pinus roxburghii forests show lower levels. Theoretical Importance: The findings highlight the importance of considering mosses in forest management plans to improve soil microbial diversity, enzymatic activity, soil quality, and health. Additionally, this research contributes to understanding the role of lower plants, such as mosses, in influencing ecosystem dynamics. Conclusion: The study concludes that moss cover during the rainy season significantly influences soil microbial biomass and enzymatic activity. Quercus and Cupressus torulosa dominated forests demonstrate higher levels of SMBC and enzymatic activity, indicating the importance of these forest types in sustaining soil microbial diversity and soil health. Including mosses in forest management plans can improve soil quality and overall ecosystem dynamics.

Keywords: moss cover, seasons, soil enzymes, soil microbial biomass, temperate forest types

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Authors: Abhichet Nobhiwong, Jiraporn Rojtinnakorn, Udomluk Sompong

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Six probiotic strains isolated from Chiang Mai and Chiang Rai province, Thailand; CR1-2, CM3-4, CM5-2, CR7-8, CM10-5 and CM10-8 were used to study their morphology and inhibition activity on three pathogenic bacteria; Aeromonas sp., Streptococcus sp. and Flavobacterium sp. that isolated from infected Nile tilapia. The agar well diffusion technique was applied for 24 and 48 hours incubation. Interestingly, some probiotics showed good inhibition activity both 24 and 48 hours on each 3 bacterial pathogens. The capable inhibiting Aeromonas sp. were CR1-2 and CR5-2 with inhibition diameters of 13.0 mm and 11.2 mm, respectively. For Streptococcus sp., effective probiotics were CR10-2 with inhibition diameters of 10.7 mm. Whereas for Flavobacterium sp., effective probiotics were CR5-2 with inhibition diameter of 9.7 mm. It can be concluded that these probiotics have potentiality to develop as the pathogens biocontrol products. These will be support for safety and organic aquaculture that which the most worthy for people health.

Keywords: probiotics, Aeromanas sp., Streptococcus sp., Flavobacterium sp.

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Authors: Yenny Paola Morales Cortés, Fabián Rico Rodríguez, Juan Carlos Serrato Bermúdez, Carlos Arturo Martínez Riascos

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Numerous benefits of galactooligosaccharides (GOS) as prebiotics have motivated the study of enzymatic processes for their production. These processes have special complexities due to several factors that make difficult high productivity, such as enzyme type, reaction medium pH, substrate concentrations and presence of inhibitors, among others. In the present work the production of galactooligosaccharides (with different degrees of polymerization: two, three and four) from lactose was studied. The study considers the formulation of a mathematical model that predicts the production of GOS from lactose using the enzyme β-galactosidase. The effect of pH in the reaction was studied. For that, phosphate buffer was used and with this was evaluated three pH values (6.0.6.5 and 7.0). Thus it was observed that at pH 6.0 the enzymatic activity insignificant. On the other hand, at pH 7.0 the enzymatic activity was approximately 27 times greater than at 6.5. The last result differs from previously reported results. Therefore, pH 7.0 was chosen as working pH. Additionally, the enzyme concentration was analyzed, which allowed observing that the effect of the concentration depends on the pH and the concentration was set for the following studies in 0.272 mM. Afterwards, experiments were performed varying the lactose concentration to evaluate its effects on the process and to generate the data for the adjustment of the mathematical model parameters. The mathematical model considers the reactions of lactose hydrolysis and transgalactosylation for the production of disaccharides and trisaccharides, with their inverse reactions. The production of tetrasaccharides was negligible and, because of that, it was not included in the model. The reaction was monitored by HPLC and for the quantitative analysis of the experimental data the Matlab programming language was used, including solvers for differential equations systems integration (ode15s) and nonlinear problems optimization (fminunc). The results confirm that the transgalactosylation and hydrolysis reactions are reversible, additionally inhibition by glucose and galactose is observed on the production of GOS. In relation to the production process of galactooligosaccharides, the results show that it is necessary to have high initial concentrations of lactose considering that favors the transgalactosylation reaction, while low concentrations favor hydrolysis reactions.

Keywords: β-galactosidase, galactooligosaccharides, inhibition, lactose, Matlab, modeling

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Authors: Sonia Mbarki Marian Brestic, Artemio Cerda Naceur Jedidi, Jose Antonnio Pascual Chedly Abdelly

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Monitoring the effect addition bio-amendment as compost to an agricultural soil for growing plant lolium perenne irrigated with a CdCl2 solution at 50 µM on physicochemical soils characteristics and plant production in laboratory condition. Even microbial activity indexes (acid phosphatase, β-glucosidase, urease, and dehydrogenase) was determined. Basal respiration was the most affected index, while enzymatic activities and microbial biomass showed a decrease due to the cadmium treatments. We noticed that this clay soil with higher pH showed inhibition of basal respiration. Our results provide evidence for the importance of ameliorating effect compost on plant growth even when soil was added with cadmium solution at 50 µmoml.l-1. Soil heavy metal concentrations depended on heavy metals types, increased substantially with cadmium increase and with compost addition, but the recorded values were below the toxicity limits in soils and plants except for cadmium.

Keywords: compost, enzymatic activity, lolium perenne, bioremediation

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Authors: J. B. Minari, O. B. Oloyede

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Polymorphonuclear neutrophils (PMNs) play a crucial role in a variety of infections caused by bacteria, fungi, and parasites. Indeed, the involvement of PMNs in host defence against Plasmodium falciparum is well documented both in vitro and in vivo. Many of the antimalarial drugs such as chloroquine used in the treatment of human malaria significantly reduce the immune response of the host in vitro and in vivo. Myeloperoxidase is the most abundant enzyme found in the polymorphonuclear neutrophil which plays a crucial role in its function. This study was carried out to investigate the effect of chloroquine on the enzyme. In investigating the effects of the drug on myeloperoxidase, the influence of concentration, pH, partition ratio estimation and kinetics of inhibition were studied. This study showed that chloroquine is concentration-dependent inhibitor of myeloperoxidase with an IC50 of 0.03 mM. Partition ratio estimation showed that 40 enzymatic turnover cycles are required for complete inhibition of myeloperoxidase in the presence of chloroquine. The influence of pH on the effect of chloroquine on the enzyme showed significant inhibition of myeloperoxidase at physiological pH. The kinetic inhibition studies showed that chloroquine caused a non-competitive inhibition with an inhibition constant Ki of 0.27mM. The results obtained from this study shows that chloroquine is a potent inhibitor of myeloperoxidase and it is capable of inactivating the enzyme. It is therefore considered that the inhibition of myeloperoxidase in the presence of chloroquine as revealed in this study may partly explain the impairment of polymorphonuclear neutrophil and consequent immunosuppression of the host defence system against secondary infections.

Keywords: myeloperoxidase, chloroquine, inhibition, neutrophil, immune

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Authors: Zohreh Bayat, Dariush Minai-Tehrani

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Lipases constitute one of the most important groups of industrial enzymes that catalyze the hydrolysis of triacylglycerol to glycerol and fatty acids. Muscarinic antagonist relieves smooth muscle spasm of the gastrointestinal tract and effect on the cardiovascular system. In this research, the effect of a muscarinic antagonist on the lipase activity of Pseudomonas aeruginosa was studied. Lineweaver–Burk plot showed that the drug inhibited the enzyme by competitive inhibition. The IC50 value (60 uM) and Ki (30 uM) of the drug revealed the drug bound to the enzyme with high affinity. Determination of enzyme activity in various pH and temperature showed that the maximum activity of lipase was at pH 8 and 60°C both in presence and absence of the drug.

Keywords: bacteria, inhibition, kinetics, lipase

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Authors: Bhavini Patel, Aslihan Ugun-Klusek, Ellen Billet

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The two main contributing factors to familial and idiopathic form of Parkinson’s disease (PD) are oxidative stress and altered proteolysis. Monoamine oxidase-A (MAO-A) plays a significant role in redox homeostasis by producing reactive oxygen species (ROS) via deamination of for example, dopamine. The ROS generated induces chemical modification of proteins resulting in altered biological function. The ubiquitin-proteasome system, which consists of three different types or proteolytic activity, namely “chymotrypsin-like” activity (CLA), “trypsin-like” activity (TLA) and “post acidic-like” activity (PLA), is responsible for the degradation of ubiquitinated proteins. Defects in UPS are known to be strongly correlated to PD. Herein, the effect of ROS generated by MAO-A on proteasome activity and the effects of proteasome inhibition on MAO-A protein levels in WT, mock and MAO-A overexpressed (MAO-A+) SHSY5Y neuroblastoma cell lines were investigated. The data in this study report increased proteolytic activity when MAO-A protein levels are significantly increased, in particular CLA and PLA. Additionally, 20S proteasome inhibition induced a decrease in MAO-A levels in WT and mock cells in comparison to MAO-A+ cells in which 20S proteasome inhibition induced increased MAO-A levels to be further increased at 48 hours of inhibition. This study supports the fact that MAO-A could be a potential pharmaceutical target for neuronal protection as data suggests that endogenous MAO-A levels may be essential for modulating cell death and survival.

Keywords: monoamine oxidase, neurodegeneration, Parkinson's disease, proteasome

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Authors: Hamdi Belfeki, Belgacem Chandoul, Mnasser Hassouna, Mondher Mejri

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Aqueous and ethanolic extracts of eight Tunisian aromatic and medicinal plants (TAMP) were characterized by studying their composition in polyphenols and also their antiradical and antioxidant capacities. In absence and in the presence of the various extracts, α-amylase from Bacillus subtlis activity, was measured in order to detect a potential inhibition. The total contents of polyphenols and flavonoid vary in function of TAMP and the mobile phase used for the extraction (distilled water or ethanol). The ethanolic extracts showed the most significant antiradical and antioxidant activities. Only the extracts from Coriandrum sativum showed a significant inhibiting effect on the α-amylase activity. This inhibiting capacity could be correlated with the chemical profile of the two extracts, due to the fact that they have the greatest amount of total flavonoid. The ethanolic extract has the most important antioxidant and anti-radicalizing activities among the sixteen extracts studied. The inhibition kinetics of the two coriander extracts were evaluated by pre-incubation method, using Lineweaver-Burk’s equation, obtained by linearization of Michaeilis-Menten’s expression. The results showed that both extracts exercised a competitive inhibition mechanism.

Keywords: α-amylase, antioxidant activity, aromatic and medicinal plants, inhibition

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Authors: Sadia Roshan, Kulsoom Sughra, Shazia Shamas, Shamaila Irum, Haleema Sadia

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To evaluate synergistic medicinal effects of Withania somnifera (Ashwaghandha) and Murraya koenigii (Curry pata) in vitro. Antimicrobial activity was determined using the disc diffusion method against five bacterial and two fungal strains. The antioxidant activity was evaluated by the DPPH assay. The antidiabetic activity was evaluated by alpha-glucosidase inhibition assay and alpha-amylase inhibition assay. Synergistic antibacterial activity was observed against all the strains of bacteria, either Gram-positive or Gram-negative and fungi under study conditions. The maximum antibacterial activity was displayed by combined extract against E. coli i.e. 26±0.4mm. Maximum antifungal activity was shown by combined extract against Aspergillus niger, i.e., 17.3±0.5mm. The antioxidant activity of the combined extract was also significant. Alpha-glucosidase inhibition and alpha-amylase inhibition assays also showed synergism. Results indicate that Withania somnifera and Murraya koengii have medicinal properties. The combined extract of both plants is more potent than their individual extracts, suggesting that these can work in synergism. The research suggests that different plant extracts could be used in combination to increase their medicinal activities by many folds, thus giving an insight into future use of herbal medication.

Keywords: withania somnifera, murraya koenigii, antimicrobial activity, gram-positive bacetria, gram-negative bacteria

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Authors: Rini Jarial, Puranjan Mishra, Lakhveer Singh, Sveta Thakur, A. W. Zularisam, Mimi Sakinah

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The aim of the present study was to assess the biological properties of Camellia sinensis and to identify its functional compounds. The methanolic leaves-extract (MLE) of commercial green tea (Camellia sinensis) was assessed for anti-bacterial activities by measuring inhibition zones against a panel of pathogenic bacterial strains using agar diffusion method. The flavonoid (5.0 to 8.0 mg/ml) and protein content (10 to 15 mg/ml) of the MLE were recorded. MLE at a concentration of 25 μg/ml showed marked anti-bacterial activity against all bacterial strains (11-30 mm zone of inhibition) and was maximum against Staphylococcus aureus (30 mm). The MLE of Camellia sinensis had the best MIC values of 2.25 and 0.56 mg/ml against S. aureus and Enterobacter sp., respectively. The MLE also possessed good anti-lipolytic activity (65%) against a Porcine pancreatic lipase (PPL) and cholesterol oxidase inhibition (79%). The present study provided strong experimental evidences that the MLE of Camellia sinensis is not only a potent source of natural anti-oxidants and anti-bacterial activity but also possesses efficient cholesterol degradation and anti-lipolytic activities that might be beneficial in the body weight management.

Keywords: anti-oxidant, anti-bacterial activity, anti-lipolytic activity, Camellia sinensis, phyto-chemicals

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Authors: M. E. Abalaka, O. A. Falusi, M. Galadima, D. Damisa

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The antimicrobial activity of aqueous, ethanolic and methanolic extracts of Chromolaena odorata (Siam weed) was evaluated against four wound isolates: Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae at the concentrations of 200mg/ml, 100mg/ml, 50mg/ml and 25mg/ml respectively. S. aureus and E. coli showed high susceptibility to the various extracts than the other test isolates. The aqueous extract showed activity against Staphylococcus aureus with a mean diameter of zone of inhibition of 16 ± 3.00 at concentration of 200mg/ml and as low as 8 ± 0.00 at concentration of 25mg/ml; E. coli showed susceptibility with a mean diameter of zone of inhibition of 18 ± 2.00 and 10 ± 0.00 at a concentration of 200mg/ml and 25mg/ml respectively. Pseudomonas aeruginosa and Klebsiella pneumoniae were resistant to the aqueous extract. Methanol extract showed activity against Staphylococcus aureus with a mean diameter of zone of inhibition at 28 ± 4.00 and 12 ± 2.30 at a concentration of 200mg/ml and 25mg/ml respectively; while E. coli was susceptible with mean diameter of zone of inhibition of 18 ± 2.00 and as low as 12 ± 0.00 at a concentration of 200mg/ml and 50mg/ml respectively, Pseudomonas aeruginosa showed considerable susceptibility with mean diameter of zone of inhibition of 13 ± 1.00 and 12 ± 0.00 at a concentration of 200mg/ml and 100mg/ml respectively. The ethanol extract showed activity against S. aureus with a mean diameter zone of inhibition of 15 ± 2.00 and 9 ± 0.00 at a concentration of 200mg/ml and 25mg/ml respectively: E. coli showed susceptibility with a mean diameter zone of inhibition of 20 ± 4.00 and 13 ± 2.00 at a concentration of 200mg/ml and 25mg/ml respectively. Pseudomonas aeruginosa showed considerable susceptibility with a mean diameter zone of inhibition of 13 ± 1.00 and 9 ± 0.00 at a concentration of 200mg/ml and 100mg/ml respectively. The results above indicate the efficacy and potency of the crude extracts of Chromolaena odorata leaf on the tested wound isolates.

Keywords: antibacterial, Chromolaena odorata, leaf extracts, test isolates

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Authors: Michelle Belinda S. Weerawardana, Gobika Thiripuranathar, Priyani A. Paranagama

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Most of fresh vegetables and fruits available in the retail markets undergo a physiological disorder in its appearance and coloration, which indeed discourages consumer purchase. A loss of millions of dollars yearly to the food industry had been due to this pronounced color reaction called Enzymatic Browning which is driven due to the catalytic activity by an oxidoreductase enzyme, polyphenol oxidase (PPO). The enzyme oxidizes the phenolic compounds which are abundantly available in fruits and vegetables as substrates into quinones, which could react with proteins in its surrounding to generate black pigments, called melanins, which are highly UV-active compounds. Annona muricata (Katu anoda) and Musa acuminata (Ash plantains) is a fruit and a vegetable consumed by Sri Lankans widely due to their high nutritional values, medicinal properties and economical importance. The objective of the present study was to evaluate and determine the effective natural anti-browning inhibitors that could prevent PPO activity in the selected fruit and vegetable. Enzyme extracts from Annona muricata (Katu anoda) and Musa acuminata (Ash plantains), were prepared by homogenizing with analytical grade acetone, and pH of each enzyme extract was maintained at 7.0 using a phosphate buffer. The extracts of inhibitors were prepared using powdered ginger rhizomes and essential oil from the bark of Cinnamomum zeylanicum. Water extracts of ginger were prepared and the essential oil from Ceylon cinnamon bark was extracted using steam distillation method. Since the essential oil is not soluble in water, 0.1µl of cinnamon bark oil was mixed with 0.1µl of Triton X-100 emulsifier and 5.00 ml of water. The effect of each inhibitor on the PPO activity was investigated using catechol (0.1 mol dm-3) as the substrate and two samples of enzyme extracts prepared. The dosages of the prepared Cinnamon bark oil, and ginger (2 samples) which were used to measure the activity were 0.0035 g/ml, 0.091 g/ml and 0.087 g/ml respectively. The measurements of the inhibitory activity were obtained at a wavelength of 525 nm using the UV-visible spectrophotometer. The results evaluated thus revealed that % inhibition observed with cinnamon bark oil, and ginger for Annona muricata was 51.97%, and 60.90% respectively. The effects of cinnamon bark oil, and ginger extract on PPO activity of Musa acuminata were 49.51%, and 48.10%. The experimental findings thus revealed that Cinnamomum zeylanicum bark oil was a more effective inhibitor for PPO enzyme present in Musa acuminata and ginger was effective for PPO enzyme present in Annona muricata. Overall both the inhibitors were proven to be more effective towards the activities of PPO enzyme present in both samples. These inhibitors can thus be corroborated as effective, natural, non-toxic, anti-browning extracts, which when added to the above fruit and vegetable will increase the shelf life and also the acceptance of the product by the consumers.

Keywords: anti-browning agent, enzymatic browning, inhibitory activity, polyphenol oxidase

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Authors: Miguel Pereira, Lígia Pimentel, Manuela Pintado

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Animal blood is a major by-product of slaughterhouses and still represents a cost and environmental problem in some countries. To be eliminated, blood should be stabilised by cooking and afterwards the slaughterhouses must have to pay for its incineration. In order to reduce the elimination costs and valorise the high protein content the aim of this study was the optimization of hydrolysis conditions, in terms of enzyme ratio and time, in order to obtain hydrolysates with biological activity. Two enzymes were tested in this assay: pepsin and proteases from Cynara cardunculus (cardosins). The latter has the advantage to be largely used in the Portuguese Dairy Industry and has a low price. The screening assays were carried out in a range of time between 0 and 10 h and using a ratio of enzyme/reaction volume between 0 and 5%. The assays were performed at the optimal conditions of pH and temperature for each enzyme: 55 °C at pH 5.2 for cardosins and 37 °C at pH 2.0 for pepsin. After reaction, the hydrolysates were evaluated by FPLC (Fast Protein Liquid Chromatography) and tested for their antioxidant activity by ABTS method. FPLC chromatograms showed different profiles when comparing the enzymatic reactions with the control (no enzyme added). The chromatogram exhibited new peaks with lower MW that were not present in control samples, demonstrating the hydrolysis by both enzymes. Regarding to the antioxidant activity, the best results for both enzymes were obtained using a ratio enzyme/reactional volume of 5% during 5 h of hydrolysis. However, the extension of reaction did not affect significantly the antioxidant activity. This has an industrial relevant aspect in what concerns to the process cost. In conclusion, the enzymatic blood hydrolysis can be a better alternative to the current elimination process allowing to the industry the reuse of an ingredient with biological properties and economic value.

Keywords: antioxidant activity, blood, by-products, enzymatic hydrolysis

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Authors: Ozlem Temiz-Arpaci, Meryem Tasci, Fatma Sezer Senol, İlkay Erdogan Orhan

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Alzheimer’s disease (AD), a neurodegenerative disorder characterized by a progressive deterioration of memory and cognition, occurs more frequently in elderly people. Current treatment approaches in this disease with the major therapeutic strategy are based on the AChE and BChE inhibition. On the other hand, tyrosinase inhibition has become a target for the treatment of Parkinson’s disease (PD) since this enzyme may play a role in neuromelanin formation in the human brain and could be critical in the formation of dopamine neurotoxicity associated with neurodegeneration linked to PD. Also benzoxazoles are structural isosteres of natural nucleotides that can interact with biopolymers so that benzoxazoles showed a lot of different biological activities. In this study, a series of 2,5-disubstituted-benzoxazole derivatives were synthesized and were evaluated as possible inhibitors of acetylcholinesterase (AChE) / butyrylcholinesterase (BChE) and tyrosinase. The results demonstrated that the compounds exhibited a weak spectrum of AChE / BChE inhibitory activity ranging between 3.92% - 54.32% except compound 8 which showed no activity against AChE and compound 4 which showed no activity against BChE at the specified molar concentrations. Also, the compounds indicated lower than tyrosinase inhibitory activity of ranging between 8.14% - 22.90% to that of reference (kojic acid).

Keywords: AChE and BChE inhibition, Alzheimer’s disease, benzoxazoles, tyrosinase inhibition

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Authors: Nassim Belkacem, Amina Azzam, Dalila Haouchine, Kahina Bennacer, Samira Soufit

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Objective: To evaluate the antioxidant activity of the methanolic extract along with the antimicrobial activity of the essential oil of the aerial parts of Rosmarinus officinalis L. collected in the region of Bejaia (northern center of Algeria). Materials and methods: The polyphenols and flavonoids contents of the methanolic extract were measured. The antioxidant activity was evaluated using two methods: the ABTS method and DPPH assay. The antimicrobial activity was studied by the agar diffusion method against five bacterial strains (Three Gram positive strains and two Gram negative strains) and one fungus. Results: The total polyphenol and flavonoid content was about 43.8 mg gallic acid equivalent per gram (GA Eq/g) and 7.04 mg quercetin equivalent per gram (Q Eq/g), respectively. In the ABTS assay, the rosemary extract has shown an inhibition of 98.02% at the concentration of 500ug/ml with a half maximal inhibitory concentration value (IC50) of 194.92ug/ml. The results of DPPH assay have shown that the rosemary extract has an inhibition of 94.67 % with an IC50 value of 17.87ug/ml, which is lower than that of Butylhydroxyanisol (BHA) about 6.03ug/ml and ascorbic acid about 1.24μg/ml. The yield in essential oil of rosemary obtained by hydrodistillation was 1.42%. Based on the determination of the diameter of inhibition, different antimicrobial activity of the essential oil was revealed against the six tested microbes. Escherichia coli from the University Hospital (UH), Streptococcus aureus (UH) and Pseudomonas aeruginosa ATCC have a minimum inhibitory concentration value (MIC) of 62.5µl/ml. However, Bacillus sp (UH) and Staphylococcus aureus ATCC have an MIC value of 125μl/ml. The inhibition zone against Candida sp was about 24 mm. The aromatograms showed that the essential oil of rosemary exercises an antifungal activity more important than the antibacterial one.

Keywords: Rosmarinus officinalis L., maceration, essential oil, antioxidant, antimicrobial activity

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Authors: John Carlo Combista, Jimbert Tan

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The orchid, Vanda tessellata was chosen by the researchers because of the presence of the constituents in the family Orchidaceae such as alkaloids, flavonoids and glycosides that might give an inhibition activity of the carbonic anhydrase enzyme. This study aimed to determine the in vitro inhibition of carbonic anhydrase of Vanda tessellata flower extract. With the use of modified colorimetric Maren T.H. (1960) method, the time in seconds each test solution changed its color after the rate of CO2 hydration were recorded. Two solvents were used: the semi-polar, 95% ethanol and the non-polar, dichloromethane solvents. The percent inhibition activity of carbonic anhydrase of the different concentrations of solvents ethanol (1%, 25% and 50%) and dichloromethane (1% and 10%) test solutions were determined. Results showed that the ethanol-based extract of Vanda tessellata in different concentrations showed an inhibitory effect while the dichloromethane-based extract of Vanda tessellata showed no inhibitory effect of carbonic anhydrase activity. For ethanol extract, the concentration with the highest activity was 50% followed by 25% which changed its color from red to yellow with an average time of 13.11 seconds and 11.57 seconds but 1% with an average time of 7.56 seconds did not exhibit an effect. The researchers recommend the isolation of the specific active constituents of Vanda tessellata that is responsible for the inhibitory effect of carbonic anhydrase enzyme. It is also recommended to utilize different blood types to observe different reactions to the inhibition of the carbonic anhydrase.

Keywords: carbonic anhydrase, inhibition, modified colorimetric Maren TH method, Vanda orchid

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Authors: F. Belkhiri, A. Baghiani, S. Boumerfeg, N. Charef, S. Khennouf, L. Arrar

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The present study was conducted to evaluate the in vitro antioxidant and antibacterial properties of methanolic extracts from roots of Tamus communis L. (TCRE), which is a plant used in traditional medicine in Algeria. The antioxidant potential of pattern was evaluated using tow complementary techniques, inhibition of free radical DPPH and the test of β-Carotene/linoleic acid. The antioxidant test indicates that non-polar fractions of TCRE (chloroform and ethyl acetate fractions) were more active than the polar fractions. Among these fractions, the chloroform extract appear in the DPPH test an IC50 of (18.89 µg/ml) comparable to that of BHT (18.6 µg/ml). This fraction was able to inhibiting the oxidation of β-Carotene with a percentage of inhibition (89.84 %). In antibacterial test, non-polar fractions showed antibacterial activity very important compared with the polar fractions. These fractions have inhibited the growth of four from nine bacterial strains, causing zones of inhibition from 08 to 23 mm of diameter.

Keywords: antioxidant activity, antibacterial activity, Tamus communis L., polar fractions

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Authors: Popy Bora, L. C. Bora, A. Bhattacharya, Sehnaz S. Ahmed

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Microbial bioagents, viz., Pseudomonas fluorescens, Bacillus subtilis, and Trichoderma viride were assessed for their ability to suppress grey blight caused by Pestalotiopsis theae, a major disease of tea crop in Assam. The expression of defense-related phytochemicals due to the application of these bioagents was also evaluated. The individual bioagents, as well as their combinations, were screened for their bioefficacy against P. theae in vitro using nutrient agar (NA) as basal medium. The treatment comprising a combination of the three bioagents, P. fluorescens, B. subtilis, and T. viride showed significantly the highest inhibition against the pathogen. Bioformulation of effective bioagent combinations was further evaluated under field condition, where significantly highest reduction of grey blight (90.30%), as well as the highest increase in the green leaf yield (10.52q/ha), was recorded due to application of the bioformulation containing the three bioagents. The application of the three bioformulation also recorded an enhanced level of caffeine (4.15%) and polyphenols (22.87%). A significant increase in the enzymatic activity of phenylalanine ammonia-lyase, peroxidase and polyphenol oxidase were recorded in the plants treated with the microbial bioformulation of the three bioagents. The present investigation indicates the role of microbial agents in suppressing disease, inducing plant defense response, as well as improving the quality of tea.

Keywords: enzymatic activity, grey blight, microbial bioagents, Pestalotiopsis theae, phytochemicals, plant defense, tea

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Authors: Imran Muhammad

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The main cause of infectious disease in the modern world is Mycobacterium Tuberculosis (MT). To combat tuberculosis, new and efficient drugs are an urgent need in the modern world. Schif bases are potent for their biological pharmacophore activity. Thus we selected different Vanillin-based Schiff bases for their binding activity against target enzymes of Mycobacterium tuberculosis that is (DprE1 (decaprenyl phosphoryl-β-D-ribose 2′-epimerase), and DNA gyrase subunit-A), using molecular docking. We evaluate the inhibition potential, interaction, and binding mode of these compounds with the target enzymes.

Keywords: schiff bases, tuberculosis, DNA gyrase, DprE1, docking

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Authors: H. Malainine, A. Amrouche, H. Benmehdi

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The aim of the present work was aimed at evaluating antifungal effect of crude esters and their corresponding FAMEs isolated from Citrullus colocynthis L., Linum usitatissimum L. and Nigella sativa L. seeds against two toxigenic fungal strains namely Aspergillus flavus and Aspergillus ochraceus. The results of the antifungal activity performed radial growth on solid medium (PDA; potatoes dextrose agar) showed that the crude esters and their corresponding FAMEs have exhibited against the two strains tested. Overall, FAMEs have provided an antifungal effect more efficient than that of crude esters. Inhibition of Aspergillus ochraceus has been labeled with percentages ranging from 13.33 to 26.61% by crude esters, While FAMEs inhibition was ranged between 27.33 to 41.13%. However, the inhibition observed against the Aspergillus flavus was varying from 14.68 to 18.59% by crude esters compared with the inhibition percentages ranging from 21.5 to 33.45% by the FAMEs. The antifungal potency of esters oils seeds of the studied plants may be an alternative for consideration by the authorities interested, due to serving the public health, in reducing the fungal enormous peril.

Keywords: Citrullus colocynthis L., Linum usitatissimum L., Nigella sativa L., FAMEs, antifungal activity, Aspergillus flavus, Aspergillus ochraceus

Procedia PDF Downloads 223