Search results for: antifungal protein

Authors: Ahmed E. Al-Sabri, Mohamed A. Moslem, Sarfaraz Hadi

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To avoid the harmful effects of the chemical fungicides on the human and minimize the environmental pollution, an alternative eco-friendly control strategies should be developed. The extract of Commiphora myhrra L. was tested against twenty fungal genera isolated from the indoor air collected from different rooms in King Saud University, Kingdom of Saudi Arabia. Disc diffusion test was modified for use in this study and the collected data was statistically analyzed. Variable antifungal efficacy of different myrrh extract was recorded against the investigated fungal genera. The efficacy of the extract was increased as the concentration increased. The highest growth inhibition (74.6%) was against Acremonium strictum followed by Trichoderma psuedokoningii (70.6%). On contrast, the lowest efficacy (12.7%) was against Ulocladium consortiale. It could be concluded that myrrh extract is promised as a source of substances from which of safer and eco-friendly could be used as antimicrobial agents against a number of pathogenic fungi.

Keywords: antifungal, myrrh, antimicrobial, medicinal plant

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Authors: Nazia Zaffar, Alam Khan, Abdul Haq, Malik Badshah

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Pakistan is an agricultural country. The increasing population leads to an increase in demand for food. A large number of crops are infected by different microbes, and nutrient deficiency of soil adversely affects the yield of crops. Furthermore, the use of chemical fertilizers like Nitrogen, Phosphorus, Potassium (NPK) Urea, and Diammonium phosphate (DAP) and pesticides have environmental consequences. Therefore, there is an urgent need to explore alternative renewable and sustainable biofertilizers. Maize is one of the top growing crops in Pakistan, but it has low yield compared to other countries due to deficiency of organic matter, widespread nutrients deficiency (phosphorus and nitrogen), unbalanced use of fertilizers and various fungal diseases. In order to get rid of all these disadvantages, Digestate emerged as a win-win opportunity for the control of a few plant diseases and a replacement for the chemical fertilizers. The present study was designed to investigate the effect of Anerobic digestate on Antifungal Activity and in different parameters of Maize. The antifungal activity, minimum inhibitory concentration (MIC), and minimum fungicidal concentration (MFC) against selected phytopathogens (Colletotrichum coccodis, Pythium ultimum, Phytophthora capsci, Rhizoctonia solani, Bipolaris oryzae and Fusarium Fujikuroi) were determined by microtiter plate method. The effect of various fertilizers in different growth parameters height, diameter, chlorophyll, leaf area, biomass, and yield were studied in field experiments. The extracts from anaerobic digestate have shown antifungal activity against selected phytopathogens, the highest activity was noted against P. ultimum, the MIC activity was high in case of P. ultimum and B. oryzae. The present study concludes that anaerobic digestate have a positive effect on maize growth and yield as well as an antifungal activity which can be potentially a good biofertilizer.

Keywords: anaerobic digestate, antifungal activity, MIC, phytopathogens

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Authors: Bouchenak Fatima, Lmegharbi Abdelbaki, Houssem Degaichia, Benrebiha Fatima

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The essential oil composition of the leaf of Artemisia absinthium from region of Cherchell (The south of Algeria) was investigated by GC, GC-MS. 27 constituents were identified correspond to 84, 63% of the total oil. The major components are Thujone (60, 82%), Chamazulènel (16, 62%), ρ-cymène (4, 29%) and 2-carène (4.25%). The antimicrobial activity of oil was tested in vitro by two methods (agar diffusion and microdilution) on three plant pathogenic fungi. This oil has been tested for antimicrobial activity against three pathogenic fungi (Botrytis cinerea, Fusarium culmorum and Helminthosporium Sp.).The study of activity was evaluated by two methods: Method of diffusion in gelose and the minimum inhibitory concentration MIC. This oil exhibited an interesting antimicrobial activity. A preliminary study showed that this oil presented high toxicity against this fungus. These results, although preliminary show a good antifungal activity, to limit and inhibit stop the development of those pathogen agent.

Keywords: artemisia absinthian, extraction process, chemical study, antifungal activity

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Authors: T. C. Machaba, S. M. Mahlo

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The current study investigates the antifungal properties of crude plant extracts from selected medicinal plant species. Eight plant species used by the traditional healers and local people to treat fungal infections were selected for further phytochemical analysis and biological assay. The selected plant species were extracted with solvent of various polarities such as acetone, methanol, ethanol, hexane, dichloromethane, ethyl acetate and water. Leaf, roots and bark extracts of Maerua juncea Pax, Albuca seineri (Engl & K. Krause) J.C Manning & Goldblatt, Senna italica Mill., Elephantorrhiza elephantina (Burch.) Skeels, Indigofera circinata Benth., Schinus molle L., Asparagus buchananii Bak., were screened for antifungal activity against three animal fungal pathogens (Candida albicans, Aspergillus fumigatus and Cryptococcus neoformans). All plant extracts were active against the tested microorganisms. Acetone, dichloromethane, hexane and ethanol extracts of Senna italica and Elephantorrhiza elephantine had excellent activity against Candida albicans and A. fumigatus with the lowest MIC value of 0.02 mg/ml. Bioautography assay was used to determine the number of antifungal compounds presence in the plant extracts. No active compounds were observed in plant extracts of Indigofera circinnata, Schinus molle and Pentarrhinum insipidum with good antifungal activity against C. albicans and A. fumigatus indicating possible synergism between separated metabolites.

Keywords: antifungal activity, bioautography, ethnobotanical survey, minimum inhibitory concentration

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Authors: Ikram Mohamed Eltayeb Elsiddig, Abdel Khalig Muddather, Hiba Abdel Rahman Ali, Saad Mohamed Hussein Ayoub

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Anogeissusleiocarpus (Combretaceae) is well known for its medicinal uses in African traditional medicine, for treating many human diseases mainly skin diseases and infections.Mycetoma disease is a fungal and/ or bacterial skin infection, mainly cause by Madurella mycetomatis fungus.This study was carried out in vitro to investigate the antifungal activity of Anogeissusleiocarpus leaf extracts against the isolated pathogenicMadurellamycetomatis, by using the NCCLS modified method compared to Ketoconazole standard drug and MTT assay. The bioactive fraction was subjected to chemical analysis implementing different chromatographic analytical methods (TLC, HPLC, and LC-MS/MS). The results showed significance antifungal activity of A. leiocarpus leaf extractsagainst the isolated pathogenicM. mycetomatis, compared to negative and positive controls. The chloroform fraction showed the highest antifungal activity.The chromatographic analysis of the chloroform fraction with the highest activity showed the presence of important bioactive compounds such as ellagic and flavellagic acids derivatives, flavonoids and stilbenoid, which are well known for their antifungal activity.

Keywords: Anogeissus leiocarpus, crude extracts and fractions of Anogeissus leiocarpus, in vitrosusceptibility of Madurella mycetomatis, Madurella mycetomatis

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Authors: Guido Trautmann-Sáez, Mario Pérez-Won, Vilbett Briones, María José Bugueño, Gipsy Tabilo-Munizaga, Luis Gonzáles-Cavieres

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Shrimp by-products are a valuable source of protein. However, traditional protein extraction methods have limitations in terms of their efficiency. Protein extraction from shrimp (Pleuroncodes monodon) industrial by-products assisted with ohmic heating (OH), microwave (MW) and pulsed electric field (PEF). It was performed by chemical method (using NaOH and HCl 2M) assisted with OH, MW and PEF in a continuous flow system (5 ml/s). Protein determination, differential scanning calorimetry (DSC) and Fourier-transform infrared (FTIR). Results indicate a 19.25% (PEF) 3.65% (OH) and 28.19% (MW) improvement in protein extraction efficiency. The most efficient method was selected for the electrospinning process and obtaining fiber.

Keywords: electrospinning process, emerging technology, protein extraction, shrimp by-products

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Authors: Gulcin Yildiz, Hao Feng

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In recent years there is growing interested in using soy protein because of several advantages compared to other protein sources, such as high nutritional value, steady supply, and low cost. Soy protein isolate (SPI) is the most refined soy protein product. It contains 90% protein in a moisture-free form and has some desirable functionalities. Creating a protein-polysaccharide conjugate to be the emulsifying agent rather than the protein alone can markedly enhance its stability. This study was undertaken to examine the effects of ultrasound treatments on the physicochemical properties of SPI-starch conjugates. The soy protein isolate (SPI, Pro-Fam® 955) samples were obtained from the Archer Daniels Midland Company. Protein concentrations were analyzed by the Bardford method using BSA as the standard. The volume-weighted mean diameters D [4,3] of protein–polysaccharide conjugates were measured by dynamic light scattering (DLS). Surface hydrophobicity of the conjugates was measured by using 1-anilino-8-naphthalenesulfonate (ANS) (Sigma-Aldrich, St. Louis, MO, USA). Increasing the pH from 2 to 12 resulted in increased protein solubility. The highest solubility was 69.2% for the sample treated with ultrasonication at pH 12, while the lowest (9.13%) was observed in the Control. For the other pH conditions, the protein solubility values ranged from 40.53 to 49.65%. The ultrasound treatment significantly decreased the particle sizes of the SPI-modified starch conjugates. While the D [4,3] for the Control was 731.6 nm, it was 293.7 nm for the samples treated by sonication at pH 12. The surface hydrophobicity (H0) of SPI-starch at all pH conditions were significantly higher than those in the Control. Ultrasonication was proven to be effective in improving the solubility and emulsifying properties of soy protein isolate-starch conjugates.

Keywords: particle size, solubility, soy protein isolate, ultrasonication

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Authors: Azadeh Yasari, Michael Ganzle, Stephen Strelkov, Nuanyi Liang, Jonathan Curtis, Nat N. V. Kav

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Pathogenic fungi cause significant yield losses and quality reductions to major crops including wheat, canola, and barley. Toxic metabolites produced by phytopathogenic fungi also pose significant risks to animal and human health. Extensive application of synthetic fungicides is not a sustainable solution since it poses risks to human, animal and environmental health. Unsaturated fatty acids may provide an environmentally friendly alternative because of their direct antifungal activity against phytopathogens as well as through the stimulation of plant defense pathways. The present study assessed the in vitro and in vivo efficacy of two hydroxy fatty acids, coriolic acid and ricinoleic acid, against the phytopathogens Fusarium graminearum, Pyrenophora tritici-repentis, Pyrenophora teres f. teres, Sclerotinia sclerotiorum, and Leptosphaeria maculans. Antifungal activity of coriolic acid and ricinoleic acid was evaluated using broth micro-dilution method to determine the minimum inhibitory concentration (MIC). Results indicated that both ricinoleic acid and coriolic acid showed antifungal activity against phytopathogens, with the strongest inhibitory activity against L. maculans, but the MIC varied greatly between species. An antifungal effect was observed for coriolic acid in vivo against pathogenic fungi of wheat and barley. This effect was not correlated to the in vitro activity because ricinoleic acid with equivalent in vitro antifungal activity showed no protective effect in vivo. Moreover, neither coriolic acid nor ricinoleic acid controlled fungal pathogens of canola. In conclusion, coriolic acid inhibits some phytopathogens in vivo and may have the potential to be an effective crop protection agent.

Keywords: coriolic acid, minimum inhibitory concentration, pathogenic fungi, ricinoleic acid

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Authors: Zhazira Shemsheyeva, Zhanara Suleimenova, Olga Shemshura, Gulnaz Mombekova, Zhanar Rakhmetova

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Bacteria that colonize plant roots and promote plant growth are referred to as plant growth-promoting rhizobacteria (PGPR). They not only provide nutrients to the plants (direct plant growth promotion) and protect plants against the phytopathogens (indirect plant growth promotion) but also increase the soil fertility. Indirectly PGPRs improve the plant growth by becoming a biocontrol agent for a fungal pathogen. The antifungal activities of the PGPrhizobacteria were assayed against different species of phytopathogenic fungi such as Fusarium tricinctum, Fusarium oxysporum, Sclerotiniasclerotiorum, and Botrytis cinerea. Pseudomonas putidaSM-1, Azotobacter sp., and Bacillus thuringiensis AKS/16 strains have been used in experimental tests on growth inhibition of phytopathogenic fungi infecting Kidney beans. Agar well diffusion method was used in this study. Diameters of the zones of inhibition were measured in millimeters. It was found that Bacillus thuringiensis AKS/16 strain showed the lowest antifungal activity against all fungal pathogens tested. Zones of inhibition were 15-18 mm. In contrast, Pseudomonas putida SM-1 exhibited good antifungal activity against Fusarium oxysporum and Fusarium tricinctum by producing 29-30 mm clear zones of inhibition. The moderate inhibitory effect was shown by Azotobacter sp. against all fungal pathogens tested with zones of inhibition from24 to 26 mm. In summary, Pseudomonas putida SM-1 strain demonstrated the potential of controlling root rot diseases in kidney beans.

Keywords: PGPR, pseudomonas putida, kindey beans, antifungal activity

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Authors: Ravid Inbar

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CaMKII (calcium-calmodulin dependent protein kinase II) makes up 2% of the protein in our brain and has a critical role in memory formation and long-term potentiation of neurons. Despite this, research has yet to uncover the role of one of the domains on the activation of this kinase. The following proposes to express the protein without the hub domain in E. coli, leaving only the kinase and regulatory segment of the protein. Next, a series of kinase assays will be conducted to elucidate the role the hub domain plays on CaMKII sensitivity to calcium/calmodulin activation. The hub domain may be important for activation; however, it may also be a variety of domains working together to influence protein activation and not the hub alone. Characterization of a protein is critical to the future understanding of the protein's function, as well as for producing pharmacological targets in cases of patients with diseases.

Keywords: CaMKII, hub domain, kinase assays, kinase + reg seg

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Authors: I. Nanako, Mariko Era, Hiroshi Morita

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Objectives: Japanese uses TATAMI rather than flooring at home. Igusa ( Juncus effuses var. decipiens ), which is commonly known in the forms of TATAMI. Juncus spp. grow at a relatively high humidity area (Japan, China and Southeast Asia ). Yatsushiro region in the southern part of Kumamoto prefecture is major produing area of Igusa. Igusa found to have honeycomb structure and was also shown to have the ability to control humidity. And Igusa has been used as a medicinal herb for diuretic and antiphlogistic agent. In previous study, we investigated antimicrobial effects of Igusa, and showed high antimicrobial activity against food poisoning bacteria. Therefore, the food trays blended Igusa can be kept clean by antimicrobial activity of Igusa. We focus on ‘Igusa foam materials’. In this study, we investigated the antibacterial and antifungal activity of Igusa, and new application to foam materials for food industry. Materials and method: We used Igusa foam materials (3 × 3 × 3 cm) as a sample. We set about fifteen types of samples combined with a commercial antibacterial agent A, a commercial antibacterial agent B, potassium laurate (C12K) and a commercial antifungal agent C, a commercial antifungal agent D and a commercial antifungal agent E. We selected four bacteria strains (Escherichia coli NBRC 3972, Staphylococus aureus NBRC 12732, Salmonella typhimurium NBRC 13245, Bacillus subtilis NBRC 3335 ) and three fungus strains (Penicillium pinophilum NBRC 6345, Cladosporium cladosporioides NBRC 30314, Aspergillus oryzae NBRC 5238 ). The fungus was cultured at 30 °C on Igusa foam materials after inoculation of the fungus for fourteen days. The bacteria was cultured at 30 °C on Igusa foam materials after inoculation of the bacteria for three days. And the Igusa foam materials were washed with 10 mL normal saline after three days. The normal saline washed Igusa foam materials plated the NA medium. After, It was cultured at 30 °C and used colony counting method. Result and Conclusion: The fifteen types of sample of Igusa foam materials had antifungal activity against C. cladosporioides, A. oryzae and P. pinophilum for fourteen days. The four types of sample contained potassium laurate and antibacterial agent A, sample contained antibacterial agent B and antifungal agent D, sample contained A and antifungal agent E, sample contained B and E had antibacterial activity against B. subtilis. The three types of sample contained potassium laurate and A, sample contained B and D, sample contained A and E had antibacterial activity against S. typhimurium. The five types of sample contained potassium laurate and A, sample contained B and D, sample contained A and E, sample contained B and E, sample contained B and antifungal agent C had antibacterial activity against E. coli and S. aureus. These results indicate that Igusa of Igusa foam materials had high antifungal activity. In addition, Igusa foam materials combined with a commercial antibacterial agent had antibacterial activity. In the future, we consider that use of Igusa foam materials may be spread from food industry.

Keywords: antibacterial, antifungal, foam materials, Igusa

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Authors: Yichao Liang, Biye Chen, Xiang Li, Steven R. Dimler

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This study investigated the effects of adding calcium and magnesium chloride on heat and storage stability of milk protein concentrate-soy protein isolate (8:2 respectively) mixtures containing 10% w/w total protein subjected to the in-container sterilization (115 °C x 15 min). The particle size does not change when emulsions are heated at pH between 6.7 and 7.3 irrespective of the mixed protein ratio. Increasing concentration of divalent cation salts resulted in an increase in protein particle size, dry sediment formation and sediment height and a decrease in pH, heat stability and hydration in milk protein concentrate-soy protein isolate mixtures solutions on sterilization at 115°C. Fortification of divalent cation salts in milk protein concentrate-soy protein isolate mixture solutions resulted in an accelerated protein sedimentation and two unique sediment regions during accelerated storage stability testing. Moreover, the heat stability decreased upon sterilization at 115°C, with addition of MgCl₂ causing a greater increase in sedimentation velocity and compressibility than CaCl₂. Increasing pH value of protein milk concentrate-soy protein isolate mixtures solutions from 6.7 to 7.2 resulted in an increase in viscosity following the heat treatment. The study demonstrated that the type and concentration of divalent cation salts used strongly impact heat and storage stability of milk protein concentrate-soy protein isolate mixture nutritional beverages.

Keywords: divalent cation salts, heat stability, milk protein concentrate, soy protein isolate, storage stability

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Authors: Mehrnaz Aminifar

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The relation between textural parameters and casein network on release of aromatic compounds was investigated over 90-days of ripening. Low DE maltodextrin and WPI were used to modify the textural properties of low fat brined cheese. Hardness, brittleness and compaction of casein network were affected by addition of maltodextrin and WPI. Textural properties and aroma release from cheese texture were affected by interaction of WPI protein-cheese protein and maltodexterin-cheese protein.

Keywords: aroma release, brined cheese, maltodexterin, WPI

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Authors: C. E. Chinma, S. O. Azeez, J. C. Anuonye, O. B. Ocheme, C. M. Yakubu, S. James, E. U. Ohuoba, I. A. Baba

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There is growing interest in the use of rice bran protein in food formulation due to its hypoallergenic protein, high nutritional value and health promoting potentials. For the first time, the amino acid profile, protein digestibility, antioxidant, and functional properties of protein concentrate from some local varieties of rice bran from Nigeria were studied for possible food applications. Protein concentrates were prepared from rice bran and analysed using standard methods. Results showed that protein content of Kwandala, Yardass, Jeep, and Jamila were 69.24%, 69.97%, 68.73%, and 71.62%, respectively while total essential amino acid were 52.71, 53.03, 51.86, and 55.75g/100g protein, respectively. In vitro protein digestibility of protein concentrate from Kwandala, Yardass, Jeep and Jamila were 90.70%, 91.39%, 90.57% and 91.63% respectively. DPPH radical inhibition of protein from Kwandala, Yardass, Jeep, and Jamila were 48.15%, 48.90%, 47.56%, and 53.29%, respectively while ferric reducing ability power were 0.52, 0.55, 0.47 and 0.67mmol TE per gram, respectively. Protein concentrate from Jamila had higher onset (92.57oC) and denaturation temperature (102.13oC), and enthalpy (0.72J/g) than Jeep (91.46oC, 101.76oC, and 0.68J/g, respectively), Kwandala (90.32oC, 100.54oC and 0.57J/g, respectively), and Yardass (88.94oC, 99.45oC, and 0.51J/g, respectively). In vitro digestibility of protein from Kwandala, Yardas, Jeep, and Jamila were 90.70%, 91.39%, 90.57% and 91.63% respectively. Oil absorption capacity of Kwandala, Yardass, Jeep, and Jamila were 3.61, 3.73, 3.40, and 4.23g oil/g sample respectively, while water absorption capacity were 4.19, 4.32, 3.55 and 4.48g water/g sample, respectively. Protein concentrates had low bulk density (0.37-0.43g/ml). Protein concentrate from Jamila rice bran had the highest foam capacity (37.25%), followed by Yardass (34.20%), Kwandala (30.14%) and Jeep (28.90%). Protein concentrates showed low emulsifying and gelling capacities. In conclusion, protein concentrate prepared from these local rice bran varieties could serve as functional ingredients in food formulations and for enriching low protein foods.

Keywords: rice bran protein, amino acid profile, protein digestibility, antioxidant and functional properties

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Authors: Prajna Metta, Yanti Rusyanti, Nunung Rusminah, Bremmy Laksono

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The decrease of neutrophil chemotaxis function may cause increased susceptibility to aggressive periodontitis (AP). Neutrophil chemotaxis is affected by formyl peptide receptor 1 (FPR1), which when activated will respond to bacterial chemotactic peptide formyl methionyl leusyl phenylalanine (FMLP). FPR1 protein value is decreased in response to a wide number of inflammatory stimuli in AP patients. This study was aimed to assess the alteration of FPR1 protein value in AP patients and if FPR1 protein value could be used as an indicator of neutrophil chemotaxis dysfunction in AP. This is a case control study with 20 AP patients and 20 control subjects. Three milliliters of peripheral blood were drawn and analyzed for FPR1 protein value with ELISA. The data were statistically analyzed with Mann-Whitney test (p>0,05). Results showed that the mean value of FPR1 protein value in AP group is 0,353 pg/mL (0,11 to 1,18 pg/mL) and the mean value of FPR1 protein value in control group is 0,296 pg/mL (0,05 to 0,88 pg/mL). P value 0,787 > 0,05 suggested that there is no significant difference of FPR1 protein value in both groups. The present study suggests that FPR1 protein value has no significance alteration in AP patients and could not be used as an indicator of neutrophil chemotaxis dysfunction.

Keywords: aggressive periodontitis, chemotaxis dysfunction, FPR1 protein value, neutrophil

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Authors: Phakamas Rachamontree, Theerawut Phusantisampan, Natthakorn Woravutthikul, Peerapong Pornwongthong, Malinee Sriariyanun

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A total of 115 yeast strains isolated from local cassava processing wastes were measured for crude protein content. Among these strains, the strain MSY-2 possessed the highest protein concentration (>3.5 mg protein/mL). By using molecular identification tools, it was identified to be a strain of Pichia kudriavzevii based on similarity of D1/D2 domain of 26S rDNA region. In this study, to optimize the protein production by MSY-2 strain, Response Surface Methodology (RSM) was applied. The tested parameters were the carbon content, nitrogen content, and incubation time. Here, the value of regression coefficient (R2) = 0.7194 could be explained by the model, which is high to support the significance of the model. Under the optimal condition, the protein content was produced up to 3.77 g per L of the culture and MSY-2 strain contain 66.8 g protein per 100 g of cell dry weight. These results revealed the plausibility of applying the novel strain of yeast in single-cell protein production.

Keywords: single cell protein, response surface methodology, yeast, cassava processing waste

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Authors: Seyed Mohammad Hosein Al Omrani Nejad, Ali Rezvani Aghdam

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This study was done in order to evaluation different irrigation intervals on amount of protein, and gel production in Aloe vera, a traditional medicinal plant. Plants was plnted in Greenhouse and irrigated according to Accumulative Pan Evaporation(APE). The treatments were included 20, 40, 60, 80, 100, 120, 140, 160, 180, and 200 mm APE which has been showed W1,W2, W3, W4, W5, W6, W7, W8,W9 and W10 respectively.The amount of protein and gel production was measured seperately. Results showed that highest protein and fresh weight of gel obtained plants which irrigated W6 and W7 respectively. According to these results can recomend which if plant irrigatedwhen APE reached 120 and 140 mm by Class A Evaporation Pan method gel production and protein would besuitable in north of khozestan province in limited irrigation conditions.

Keywords: irrigation, protein, gel, aloe vera, Iran

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Authors: Er-Yuan Chuang

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Bio-mimetic matters have biological functionalities. This might be valuable in the development of versatile biomaterials. At biological fields, protein-based materials might be components to form a 3D network of extracellular biomolecules, containing growth factors. Also, the protein-based biomaterial provides biochemical and structural assistance of adjacent cells. In this study, we try to prepare protein based biomaterial, which was harvested from living animal. We analyzed it’s chemical, physical and biological property in vitro. Besides, in vivo bio-interaction of the prepared biomimetic matrix was tested in an animal model. The protein-based biomaterial has degradability and biocompatibility. This development could be used for tissue regenerations and be served as platform technologies.

Keywords: protein based, in vitro study, in vivo study, biomaterials

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Authors: Mohamed Abdullah Ahmed

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In a study of chickpea protein isolate (CPI) preparation, the wet alkaline extraction was carried out. The objectives were to determine the optimal extracting conditions of CPI and apply CPI into a sponge cake recipe to replace egg and make acceptable product. The design used in extraction was a central composite design. The response surface methodology was preferred to graphically express the relationship between extraction time and pH with the output variables of percent yield and protein content of CPI. It was noted that optimal extracting conditions were 60 min and pH 10.5 resulting in 90.07% protein content and 89.15% yield of CPI. The protein isolate (CPI) could be incorporated in cake to 20% without adversely affecting the cake physical properties such as cake hardness and sensory attributes. The higher protein content in cake was corresponding to the amount of CPI added. Therefore, adding CPI can significantly (p<0.05) increase protein content in cake. However, sensory evaluation showed that adding more than 20% of CPI decreased the overall acceptability. The results of this investigation could be used as a basic knowledge of CPI utilization in other food products.

Keywords: chick bean protein isolate, sponge cake, utilization, sponge

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Authors: Shayli Varasteh Moradi, Wayne A. Johnston, Dejan Gagoski, Kirill Alexandrov

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The demand for a rapid and more efficient technique to identify protein-protein interaction particularly in the areas of therapeutics and diagnostics development is growing. The method described here is a rapid in vitro protein-protein interaction analysis approach based on AlphaLISA technology combined with Leishmania tarentolae cell-free protein production (LTE) system. Cell-free protein synthesis allows the rapid production of recombinant proteins in a multiplexed format. Among available in vitro expression systems, LTE offers several advantages over other eukaryotic cell-free systems. It is based on a fast growing fermentable organism that is inexpensive in cultivation and lysate production. High integrity of proteins produced in this system and the ability to co-express multiple proteins makes it a desirable method for screening protein interactions. Following the translation of protein pairs in LTE system, the physical interaction between proteins of interests is analysed by AlphaLISA assay. The assay is performed using unpurified in vitro translation reaction and therefore can be readily multiplexed. This approach can be used in various research applications such as epitope mapping, antigen-antibody analysis and protein interaction network mapping. The intra-viral protein interaction network of Zika virus was studied using the developed technique. The viral proteins were co-expressed pair-wise in LTE and all possible interactions among viral proteins were tested using AlphaLISA. The assay resulted to the identification of 54 intra-viral protein-protein interactions from which 19 binary interactions were found to be novel. The presented technique provides a powerful tool for rapid analysis of protein-protein interaction with high sensitivity and throughput.

Keywords: AlphaLISA technology, cell-free protein expression, epitope mapping, Leishmania tarentolae, protein-protein interaction

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Authors: Muhammad Arshad, Ghulam Hussain Bhatti, Abdul Qayyum

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Zinc-doped silica oxide nanoparticles having size 7.93nm were synthesized by the deposition precipitation method by using different solvents (acetonitrile, n-hexane, isoamylalchol). Biological potential such as antibacterial activities against Bacillussubtilusand Escherichia coli, and antifungal activities against Candida parapsilosis and Aspergilusniger were also investigated by Disc diffusion method. Different characterizations techniques including Fournier Transmission Infrared Spectroscopy (FT-IR), X-ray diffraction (XRD), Scanning Electron Microscope (SEM), Transmission Electron Microscope (TEM), Thermo-gravimeteric Analysis (TGA), Atomic forced microscopy (AFM), and Dynamic Light Scattering (DLS) were used. FT-IR characterization confirmed the presence of metal oxide bond (SiO2) while XRD showed the hexagonal structure. SEM and TEM characterization showed the morphology of nanoparticles. AFM study showed good particle size distribution as depicted by a histogram. DLS study showed the gradual decease in the size of nanoparticles from 24.86nm to 13.24 nm. Highest antibacterial activities revealed by acetonitrile solvents (6%and 4.5%) followed by isoamylalchol (3% and 2.4%) while n-hexane solvent showed the lowest activity (2%and 1%) respectively. Higher antifungal activities exhibited by n-hexane (0.34 % and 0.43%) followed by isoamylalchol (0.27% and 0.19%) solvent while acetonitrile (0.21% and 0.17%) showed least activity respectively. Statistical analysis by using one-way ANOVA also indicated the significant results of both biological activities.

Keywords: nanoparticles, precipitation methods, antibacterial, antifungal, characterizations

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Authors: Yui Okuno, Mariko Era, Takayoshi Kawahara, Takahide Kanyama, Hiroshi Morita

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Introduction: Fungus and mite are known as allergens that cause an allergic disease for example asthma bronchiale and allergic rhinitis. Cladosporium cladosporioides is one of the most often detected fungi in the indoor environment and causes pollution and deterioration. Dermatophagoides farinae is major mite allergens indoors. Therefore, the creation of antifungal agents with high safety and the antifungal effect is required. Fatty acid salts are known that have antibacterial activities. This report describes the effects of fatty acid salts against Cladosporium cladosporioides NBRC 30314 and Dermatophagoides farinae. Methods: Potassium salts of 9 fatty acids (C4:0, C6:0, C8:0, C10:0, C12:0, C14:0, C18:1, C18:2, C18:3) were prepared by mixing the fatty acid with the appropriate amount of KOH solution to a concentration of 175 mM and pH 10.5. The antifungal method, the spore suspension (3.0×104 spores/mL) was mixed with a sample of fatty acid potassium (final concentration of 175 mM). Samples were counted at 0, 10, 60, 180 min by plating (100 µL) on PDA. Fungal colonies were counted after incubation for 3 days at 30 °C. The MIC (minimum inhibitory concentration) against the fungi was determined by the two-fold dilution method. Each fatty acid salts were inoculated separately with 400 µL of C. cladosporioides at 3.0 × 104 spores/mL. The mixtures were incubated at the respective temperature for each organism for 10 min. The tubes were then contacted with the fungi incubated at 30 °C for 7 days and examined for growth of spores on PDA. The acaricidal method, twenty D. farinae adult females were used and each adult was covered completely with 2 µL fatty acid potassium for 1 min. The adults were then dried with filter paper. The filter paper was folded and fixed by two clips and kept at 25 °C and 64 % RH. Mortalities were determained 48 h after treatment under the microscope. D. farina was considered to be dead if appendages did not move when prodded with a pin. Results and Conclusions: The results show that C8K, C10K, C12K, C14K was effective to decrease survival rate (4 log unit) of the fatty acids potassium incubated time for 10 min against C. cladosporioides. C18:3K was effective to decrease 4 log unit of the fatty acids potassium incubated time for 60 min. Especially, C12K was the highest antifungal activity and the MIC of C12K was 0.7 mM. On the other hand, the fatty acids potassium showed no acaricidal effects against D. farinae. The activity of D. farinae was not adversely affected after 48 hours. These results indicate that C12K has high antifungal activity against C. cladosporioides and suggest the fatty acid potassium will be used as an antifungal agent.

Keywords: fatty acid salts, antifungal effects, acaricidal effects, Cladosporium cladosporioides, Dermatophagoides farinae

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Authors: Suman Bala, Sunil Kamboj, Vipin Saini

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Quantitative Structure Activity Relationship (QSAR) analysis has been developed to relate antifungal activity of novel substituted 1,3,4-oxadiazole against Candida albicans and Aspergillus niger using computer assisted multiple regression analysis. The study has shown the better relationship between antifungal activities with respect to various descriptors established by multiple regression analysis. The analysis has shown statistically significant correlation with R² values 0.932 and 0.782 against Candida albicans and Aspergillus niger respectively. These derivatives were further subjected to molecular docking studies to investigate the interactions between the target compounds and amino acid residues present in the active site of glucosamine-6-phosphate synthase. All the synthesized compounds have better docking score as compared to standard fluconazole. Our results could be used for the further design as well as development of optimal and potential antifungal agents.

Keywords: 1, 3, 4-oxadiazole, QSAR, multiple linear regression, docking, glucosamine-6-phosphate synthase

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Authors: Duc Dong Do, Ngoc Ha Tran, Thanh Hai Dang, Cao Cuong Dang, Xuan Huan Hoang

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Global aligning two protein-protein interaction networks is an essentially important task in bioinformatics/computational biology field of study. It is a challenging and widely studied research topic in recent years. Accurately aligned networks allow us to identify functional modules of proteins and/ororthologous proteins from which unknown functions of a protein can be inferred. We here introduce a novel efficient heuristic global network alignment algorithm called FASTAn, including two phases: the first to construct an initial alignment and the second to improve such alignment by exerting a local optimization repeated procedure. The experimental results demonstrated that FASTAn outperformed the state-of-the-art global network alignment algorithm namely SPINAL in terms of both commonly used objective scores and the run-time.

Keywords: FASTAn, Heuristic algorithm, biological network alignment, protein-protein interaction networks

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Authors: Xiao Zhou, Jianlin Cheng

Abstract:

A single amino acid mutation can have a significant impact on the stability of protein structure. Thus, the prediction of protein stability change induced by single site mutations is critical and useful for studying protein function and structure. Here, we presented a deep learning network with the dropout technique for predicting protein stability changes upon single amino acid substitution. While using only protein sequence as input, the overall prediction accuracy of the method on a standard benchmark is >85%, which is higher than existing sequence-based methods and is comparable to the methods that use not only protein sequence but also tertiary structure, pH value and temperature. The results demonstrate that deep learning is a promising technique for protein stability prediction. The good performance of this sequence-based method makes it a valuable tool for predicting the impact of mutations on most proteins whose experimental structures are not available. Both the downloadable software package and the user-friendly web server (DNpro) that implement the method for predicting protein stability changes induced by amino acid mutations are freely available for the community to use.

Keywords: bioinformatics, deep learning, protein stability prediction, biological data mining

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Authors: Duygu Çimen, Nilay Bereli, Adil Denizli

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In this study is to synthesis magnetic nanoparticles for purify protein C. For this aim, N-Methacryloyl-(L)-histidine methyl ester (MAH) containing 2-hydroxyethyl methacrylate (HEMA) based magnetic nanoparticles were synthesized by using micro-emulsion polymerization technique for templating protein C via metal chelation. The obtained nanoparticles were characterized with Fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), zeta-size analysis and electron spin resonance (ESR) spectroscopy. After that, they were used for protein C purification from aqueous solution to evaluate/optimize the adsorption condition. Hereby, the effecting factors such as concentration, pH, ionic strength, temperature, and reusability were evaluated. As the last step, protein C was determined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Keywords: immobilized metal affinity chromatography (IMAC), magnetic nanoparticle, protein C, hydroxyethyl methacrylate (HEMA)

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Authors: Ahmed M. Hjazi, Bader M. Hjazi

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Protein 4.1 is a crucial component of complex interactions between the cytoskeleton and other junctional complex proteins. When the gene encoding this protein is altered, resulting in reduced expression, or when the protein is absent, the red cell undergoes a significant structural change. This research aims to achieve a deeper comprehension of the biochemical effects of red cell protein deficiency. A Tandem Mass Spectrometry Analysis (TMT-MS/MS) of patient cells lacking protein 4.1 compared to three healthy controls was achieved by the Proteomics Institute of the University of Bristol. The SDS-PAGE and Western blotting were utilized on the original patient sample and controls to partially confirm TMT MS/MS data analysis of the protein-4.1-deficient cells. Compared to healthy controls, protein levels in samples lacking protein 4.1 had a significantly higher concentration of proteins that probably originated from reticulocytes. This could occur if the patient has an elevated reticulocyte count. The increase in chaperone and reticulocyte-associated proteins was most notable in this study. This may result from elevated quantities of reticulocytes in patients with hereditary elliptocytosis.

Keywords: hereditary elliptocytosis, protein 4.1, red cells, tandem mass spectrometry data.

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Authors: Yusuf Ali Abdulle, Azhar Uddin Keerio

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Background: Protein elicitors play a key role in signaling or displaying plant defense mechanisms and emerging as vital tools for bio-control of insects. This study was aimed at the characterization of the novel protein elicitor isolated from entomopathogenic fungi Lecanicillium lecanii (V3) strain and its activity against Whitefly, Bemisia tabaci in cotton. The sequence of purified elicitor protein showed 100% similarity with hypothetical protein LEL_00878 [Cordyceps confragosa RCEF 1005], GenBank no (OAA81333.1). This novel protein elicitor has 253 amino acid residues and 762bp with a molecular mass of 29 kDa. The protein recombinant was expressed in Escherichia coli using pET‐28a (+) plasmid. Effects of purified novel protein elicitor on Bemisia tabaci were determined at three concentrations of protein (i.e., 58.32, 41.22, 35.41 μg mL⁻¹) on cotton plants and were exposed to newly molted adult B.tabaci. Bioassay results showed a significant effect of the exogenous application of novel protein elicitor on B. tabaci in cotton. In addition, the gene expression analysis found a significant up-regulation of the major genes associated with salicylic acid (SA) and jasmonic acid (JA) linked plant defense pathways in elicitor protein-treated plants. Our results suggested the potential application of a novel protein elicitor derived from Lecanicillium lecanii as a future bio-intensive controlling approach against the whitefly, Bemisia tabaci.

Keywords: resistance, Lecanicillium lecanii, secondary metabolites, whitefly

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Authors: Angela U. Makolo, Temitayo A. Olagunju

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The knowledge of signaling pathways is central to understanding the biological mechanisms of organisms since it has been identified that in eukaryotic organisms, the number of signaling pathways determines the number of ways the organism will react to external stimuli. Signaling pathways are studied using protein interaction networks constructed from protein-protein interaction data obtained using high throughput experimental procedures. However, these high throughput methods are known to produce very high rates of false positive and negative interactions. In order to construct a useful protein interaction network from this noisy data, computational methods are applied to validate the protein-protein interactions. In this study, a computational technique to identify signaling pathways from a protein interaction network constructed using validated protein-protein interaction data was designed. A weighted interaction graph of the Saccharomyces cerevisiae (Baker’s Yeast) organism using the proteins as the nodes and interactions between them as edges was constructed. The weights were obtained using Bayesian probabilistic network to estimate the posterior probability of interaction between two proteins given the gene expression measurement as biological evidence. Only interactions above a threshold were accepted for the network model. A pathway was formalized as a simple path in the interaction network from a starting protein and an ending protein of interest. We were able to identify some pathway segments, one of which is a segment of the pathway that signals the start of the process of meiosis in S. cerevisiae.

Keywords: Bayesian networks, protein interaction networks, Saccharomyces cerevisiae, signalling pathways

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Authors: Gül Ebru Orhun

Abstract:

A half diallel crossing design was carried out during 2011 and 2012 growing seasons under Çanakkale-Turkey ecological conditions. In this research, 20 F1 maize hybrids obtained by 6x6 half diallel crossing were used. Gene action for protein content and grain yield traits were explored in half set involving six elite inbred lines. According to the results diallel analysis dominance and additive gene variances were determined for protein content. Variance/Co-variance graphs revealed for grain yield and protein content traits. In this study, inheritance of grain yield and protein content demonstrated over-dominance type of gene action.

Keywords: protein, maize, inheritance, gene action

Procedia PDF Downloads 502