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The biocontrol activity of two isolates of Candida membranifaciens against grey mold of apple fruit caused by Botrytis mali and their ability to induce biochemical defense responses in apple tissue were investigated. Apple fruit (Malus domestica) wounds were inoculated with 50 µl yeast suspension (1 × 10⁷ CFU/ml) of C. membranifaciens followed 4 h later by 20 µl of conidial suspension of B. mali (1 × 10⁵ conidia/ml). The apples were then incubated at 20 ºC for 8 days. Lesion diameter sizes were measured 4 and 8 days after pathogen inoculation. In addition to controlling grey mold, these two isolates of C. membranifaciens caused increases in peroxidase and β-1, 3-glucanase activities. These isolates also caused inhibition in catalase activity. The accumulation of phenolic compounds was increased in apple fruit treated with antagonists and inoculated with B. mali and reached its highest level 6 days after treatment. The ability of C. membranifaciens to affect H2O2-metobolizing enzymes and increase levels of β-1, 3-glucanase activity and phenolic compounds may be some of mechanisms responsible for its biocontrol activity.  

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The effects of various levels of the commercial dietary supplement, Hoplite containing glucan, on growth performance, body composition and intestinal microbiota in white fish, Rutilus frisii kutum fry were investigated. 25 white fish fry (mean weight=1 ± 0.15 g) were stocked in each experimental tank (100 L). The fry were fed experimental diets supplemented with 0, 0.5, 1 and 2% Hoplite to apparent satiation, 3 times a day for 60 days. Biometry was performed every two weeks. Mean dissolved oxygen concentration, pH and water temperature recorded during the experiment were 5.0±0.1 mg/l, 7.8±0.2 and 24.4±0.11 ºC, respectively. At the end of the trial period, growth parameters, body composition and intestinal microbiota were studied. Results indicate that fry fed 0.5 and 1% glucan exhibited highest weight gain (WG), specific growth rate (SGR) and final body weight (FBW) which were significantly different (P

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Ganoderma lucidum is one of the best-known medicinal mushrooms in the world. It contains substantial amounts of intra- and extracellular secondary metabolites and polysaccharides each with its own specific medicinal and medical uses. The chitin-glucan complex (CGC) is considered one of the important polysaccharides of this fungus. Among the 10 various culture media that were studied, the one containing PDB at 24g/l, peptone at 1g/l, and with the dry weight of cells of 11.6 g/l, the produced CGC of 3.2g/l, and with 27.6 percent CGC in the dry weight of the cells was selected as the suitable culture medium. FTIR analysis was performed for characterization of the produced CGC and its antibacterial properties were studied. The obtained time profile for CGC growth and production was 20 days and, using the logistic growth model and the Lodding-Pipet equation, the calculated specific growth rate of Ganoderma lucidum (μm) and the volumetric productivity for the product were 2.85 g CGC L-1day-and 0.5274 day-1, respectively. The calculations indicated there were high degrees of conformance between the model and the laboratory data related to kinetic characteristics of cell growth (R2= 0.9679) and to CGC production (R2=0.9901). Therefore, the introduced kinetic model can serve as an effective guide to control the fermentation process in industrial production of the valuable CGC polymer.

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Brassica napus is an important oilseed crop and the yield loss due to fungal disease stem rot caused by Sclerotinia sclerotiorum is a serious problem in cultivation of this crop. The pathogenesis-related (PR) protein, glucanase, hydrolyzes a major cell wall component, glucan, of the pathogenic fungi and acts as a plant defense barrier. In this study, a β-1,3-glucanase (bgn13.1) gene was isolated from the biocontrol fungus Trichoderma virens-10 (showing the high β-glucanase activity) and cloned in pUC19 cloning vector. The cloned fragment was confirmed by molecular analysis and showed to contain two short introns, 52 and 57 bp and an open reading frame coding 761 amino acids. The bgn13.1 gene was over-expressed under the CaMV35S promoter in B. napus, R line Hyola 308. Transformation of cotyledonary petioles was achieved by pBIKH1 containing bgn13.1 gene via Agrobacterium tumefaciens LBA4404. The insertion of transgene was verified by the polymerase chain reaction (PCR) and genomic DNA Southern dot blotting in T0 generation. RT-PCR analysis indicated that the transgenic canola plants were able to transcribe the β-1,3 glucanase gene. Also, we used transgenic over-expression approach in order to investigate antifungal activity of expressed Bgn13.1 on S. sclerotiorum. The heterologous expressed Bgn13.1 of line # 7 and line # 10 compared with other lines showed stronger inhibition against hyphal growth of S. sclerotiorum with

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In this study, 100 fish (Astronotusocellatus) with density of 8 to 12 fish per aquarium distributed to four treatments (Control, 0/25%, 0/5% and %1 Macrogard) the experiment lasted forsix weeks. In this experiment it was observed that the number of white blood cells, especially neutrophils and serum the lysozyme level in fish fed with different levels Macrogard were significantly increased compared to the control group (p<0/05). Most changes in the white blood cell count was observed in fourth week of feeding for all Macrogard levels. This material affects the immune system of Astronotusocellatus, and we can say that the all levels Macrogard were safe. They are most effective when the fish are fed for 4 weeks with diets containing Macrogard.

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Endoglucanase Cel9A from Alicyclobacillus acidocaldarius (AaCel9A), a thermophile enzyme, randomly breaks β1-4 glycosidic bond between glucose units in cellulose polymer and produces oligosaccharides with reducing end. In this study, first of all, E.coli BL21 cells were transformed by pDEST17 carrying AaCel9A enzyme gene for expression of the recombinant enzyme. After expression, the recombinant enzyme was purified by Ni-NTA affinity chromatography column and the purity of the recombinant protein was analyzed by SDS-PAGE. Due to impact of the calcium, pH and temperature on AaCel9A activity, the effects of these parameters were investigated on AaCel9A activity to optimize activity condition by using Response surface methodology. The SDS-PAGE result showed that AaCel9A, with molecular weight of 59 kDa, was expressed and purified. Response surface methodology data reveal that the effect of pH on the activity of the enzyme is higher than temperature and the calcium effect is less than temperature. Results showed that the optimum condition of AaCel9A activity reaches at pH 6.35 and 64.5 ˚C as well as 4.92 mM of calcium. Finally, the high correlation between experimental and predicted date indicated that the proposed model for optimizing the enzyme activity has a high accuracy.

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This study aimed to isolate the thermophilic bacteria producing exoglucanase from hot springs of Dehloran, in Ilam province, in South-West of Iran. After sampling, bacterial enrichment was performed in a medium containing rice barn or CMC (carboxymethyl cellulose). Identifying bacteria was performed based on characteristics such as universal 16SrRNA gene sequencing using PCR. The isolates indicated the most similarity to species Pseudoxanthomonas mexicana، Chelatococcus daeguensis,Promicromonospora sp., Isoptericola variabilis sp. in the GenBank. The selected strain was identified and named as Isoptericola variabilis sp. IDAH9. The strain showed 1 U/ml exoglucanas activity. In order to optimization of enzyme production, the effects of carbon, nitrogen, Tween-80, and sucrose were evaluated. The results showed that the most exoglucanas activity are obtained at concentrations of 0.2% sucrose, 0.6% Tween 80 and 12 g/l of bran and carboxymethyl cellulose carbon sources and 5.6 g/l of ammonium sulfate. The residual enzyme activity was retained 64% of activity after 24 hour incubation at 50 °C. Therefore, Isoptericola variabilis sp. IDAH9 can be a suitable option for production thermostable exoglucanase from low price carbon sources..

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Cellulase enzyme has shown their potential application in different industry. cellulase immobilization is one of the different methods for enzymatic stabilization. An advantage of immobilization is enzymatic reusability, which have an economical advantage for enzyme using in industry. Properties of Chitosan as a support for enzyme immobilization are always considerable. Due to its unique biological properties such as biocompability, biodegradability and non-toxicity, chitosan is an attractive support for immobilization. In this investigation Aa-cel9A endoglucanase gene was cloned in pET28 (+) expression vector. Sequencing result had been proved gene cloning in vector. Then the constructed vector was transformed to Eshershia.Coli (BL21) cells and enzyme production was induced. The result obtained from SDS-PAGE analysis and enzymatic assay showed the recombinant protein has been expressed and protein purification was done with Ni-NTA column. Chitosan macrobeads were prepared by precipitation procedure. After immobilization of enzyme with glutaraldehyde as linker, enzyme immobilization has been proved with FTIR and Bradford analysis. The obtained result showed optimum condition for covalent immobization on support are 0.7% of glutaraldehyde concentration and sodium phosphate buffer with pH 7. Bradford analysis and enzymatic activity assay have proved 85% of enzyme molecules immobilized on support.

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Cellulase is one of the industrial enzymes which its production and utilization is increasingly taking into consideration due to global heed to second-generation bioethanol production. Cellulase produced by different organisms such as fungi, bacteria, insects, and plants. With increase in utilization of this enzyme and need for reduction in the enzymes price for production of second-generation bioethanol, the production of recombinant enzyme has been considered noticeably.
In this study, by investigation of corn steep liquor as nitrogen source and second carbon source after glycerol, a new medium is designed based on SYN6 salt medium then biomass and endoglucanase II production by methylotrophic yeast was optimized. Experiments designed by one-factor and response surface methodology used for optimization.
Results showed that optimum conditions for biomass and endoglucanase production are 5.5% (w/v) and 6.15% (w/v) of corn steep liquor respectively. New optimized conditions increased 41.4% and 69.7% for biomass and recombinant enzyme production respectively.

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In recent years, biocatalysts have widespread application in industry because they can do chemical reactions with the lowest energy and highest efficiency. Bacterial enzymes are more useful in this field due to simple cloning and expression process in the manipulated host. By considering specific role of endoglucanase enzymes in cellulose hydrolyzing reactions, these types of enzymes are more applicable in related industries. The produced glucose through enzymatic hydrolysis could be used in different industries such as biofuel and ethanol production and in the food industry as sweetener. Therefore, cloning and production of Endoglucanase in manipulated hosts has been developed in recent years. This study was performed to isolate, screen and identify native endoglucanase -producing strains from soil around the roots of the maple tree. Isolated strains were identified using 16S rRNA gene sequencing. After identifying of the bacteria (Enterobacter hormaechei), Endoglucanase enzyme gene was amplified using degenerate primers at first and then by specific primers with restriction enzymes sequences. DNA fragment and plasmid vector were treated by specific restriction enzymes and then ligated to each other. Then recombinant plasmid transferred to the E. coli BL-21 as expression host and kinetic properties of recombinant enzyme were evaluated. Expression of the target protein was done by stimulating the Lac operon by using 1 mM of IPTG and the kinetic features of the recombinant enzyme such as Vmax and Km evaluated as 45 µmol/min and 1.4 mg/ml respectively. The optimum conditions for enzyme activity tend to be 37°C at a pH of 7.

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Objective: In recent decades, β-glucans have been used as important complementary and alternative medicines for numerous immunocompromised individuals and those with end stage of cancer terminal. The most active form of β-glucans is β(1,3)D-glucan and its  most common source is cell wall of Candida albicans. Recently it has been introduced as a nano particle design to be used as a carrier for drug delivery. The current study researches a rapid method for the extraction of β(1,3)D-glucans. Methods: The present study was conducted atTarbiatModaresMedicalUniversity in 2012. Candida solubilized β-glucans were obtained by oxidation of the cell wall with sodium hypochlorite and sodium hydroxide. The particle part could be solubilized by treatment with dimethylsulfoxide (DMSO) and zymolyase digestion to extract β(1,3)D-glucan. The soluble fractions were lyophilized. We performed the Callose test to verify the presence of β(1,3)D-glucans. Solubilized fractions were dissolved in D2O and 1H-NMR spectra were measured. Results: The soluble β(1,3)D-glucan fraction which was derived from 1 g of dried Candida albicans germ tube weighed 190 mg. β(1,3)D-glucan was verified by the Callose test and ¹H-NMR test compared with Curdlan (standard). ¹H-NMR spectra verified the existence of β(1,3)D-glucan in the final product. Conclusion: In the present study, extraction of β(1,3)D-glucan by oxidation of the cell wall using sodium hypochlorite yielded more pure β(1,3)D-glucans in comparison with other extraction methods. Thus it might represent a rapid method of extraction.

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In this study, to extract beta-glucan from oats by hot water extraction method, after milling, we put it into hydrothermal process by autoclave, at three different temperatures of 106, 120 and 130 degrees Celsius in two different time (10 minutes and 20 minutes) intervals, to measure influence of time and temperature in physiochemical and functional properties of beta-glucan. The extraction process of β-glucan was accomplished by hydro extraction. The reason for choosing this extraction method was due to the higher purity of β-glucan and also has higher quality. After extraction, the physiochemical and functional properties of extracted β-glucan such as extraction efficiency, water holding capacity, emulsion capacity and stability, SEM and FTIR were tested. The result of this study showed that extracted β-glucan from hydrothermal flour had the highest extraction efficiency at 106°Ϲ in 20 minutes, had the highest emulsifier capacity at 120°Ϲ in 20 minutes (17.35%), and the highest emulsion stability at 106°Ϲ in 20 minute (19.71%), and also the highest rate of water absorption at 120°Ϲ in 10 minutes, which was 1.95%. In analyzing microstructure by Scanning Electron Microscope, it was observed that whatever the time and temperature has been increased, the b-glucan tissue would be spongy. Finally, identifying factors of the oats b- glucan by FTIR has shown in different treatments.
 

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β_glucan is dietry fibre and soluble gum and .it  has various physical properties such as thickening,stabilizing, emulsification, and gelation.in this study the effect of intensity (0,200, 300, 400watt) and time (0, 3.5  and 7minute sultrasonic waves were investigated on the physicochemical and the rheological properties of extracted.ß_glucan.The results showed that ultrasound intensity and time on moisture, ash, protein, starch and β-glucan content were significant (P <0.05), While it was not significant on fats and fibers (P >0.05), The sample with the intensity of 200 watts in 3.5 minutes showed the highest amount of β-glucan (77.1%) and the sample at 400 watts in 3.5 min with the lowest amount of ß-glucan (55.6%).Result showed that apparent viscosity decreased by increasing of the shear rate. All samples showed shear thinning behavior.The highest consistency coefficient, zero shear viscosity and lowest flow index was controlsample., and the lowest consistency coefficient and zero shear viscosity and the highest flow index was  in the sample with 400 W intensity for 3.5 min. The results obtained from fitting the data with different rheological models showed that the Herschel  andCarreau model was chosen for the best model. Linear region assign about 0.5 %using strain sweep test. Result of frequency sweep test showed first, by increasing the frequency of both the modulus of loss and storage , the storage modulus was reduced at high frequencies. The control sample was at low frequency G^²>G, at high frequency G^¢>G^², Other examples were G^¢>G^².
 

 

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ß-glucans  are non-starch polysaccharides and water soluble hydrocolloids, Apart from being nutritionally important, ß-glucans show an important technological role as thickening, stabilizing, emulsification, gel-forming component and fat substitute in the dairy products, bakeries, meats, pharmacy, cosmetics, and chemical industries, and feed production. The extraction of cereals ß-glucans is very difficult. Ultrasonic extraction, represents higher efficiency since this method is a cost- effective which shortens the extraction time ,energy and solvent consumption compared to conventional methods..in this study the effect of intensity  and time  at four different powers (0, 200, 300, 400 w)  in three time (0, 3.5 and 7 min) intrevals were investigated on the yield, recovery, color and the functional  properties of extracted ß-glucan.The results showed that the intensity and time of ultrasound were significant on yield, recovery, color, emulsion stability, water holding capacity, solubility ß-glucan (p<0.05). With increasing ultrasonic intensity and time, extracted ß-glucan yield, recovery, emulsion stability, water holding capacity increased. The highest extraction yield (3.34%), solubility (75.67%) were observed at power 400w for 7 minutes, Also the highest recovery (52.04%), emulsion stability (69.06%), water retention capacity (13.21 g /g), compared to blank sample at power 300w  for 7 minutes . L* , a^*and b* and were 76.66, 3.2, 3.2 respectively.

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The unique properties of camel milk, qualify this product to be used as a nutraceutical. In this study, functional synbiotic yogurt made from camel milk has been investigated in three levels of fat (0, 2.5 and 5% (w/v)). Probiotic bacteria (Streptococcus thermophilus, Lactobacilus delbrueckii and ssp. bulgaricus.) and β-glucan (prebiotic agent) were added in three levels of concentration (0.5, 1 and 1.5 % (v/v)) and (0, 1 and 2% (w/v)), respectively. The physicochemical properties of the product and viability of probiotic bacteria were measured on the 0, 7th and 14th days. Beta-glucan, fat and storage time had significantly (P< 0.05) increasing effects on viscosity, Water-Holding Capacity (WHC) and the viability of probiotic bacteria. These parameters caused decrease in syneresis and pH of yogurt. It was concluded that the addition of oat β-glucan to camel milk to make functional synbiotic yogurt could result in a product of acceptable physicochemical properties.

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Owning to drawbacks of synthetic polymers, biodegradable packaging materials (biopolymers) have received more attention both in studies and industrial applications. However, compared to conventional packaging, the biodegradable materials have some limitation must be eliminated either by using composite preparation method or introduction of nanotechnology. Inclusion of metal nanoparticles and their oxides are of the new approaches to improve the properties of packaging films. Therefore, the present study attempted to investigate the effect of ZnO (0, 2.5 and 5%) and β-glucan (0, 10 and 20%) incorporation on chemical and microbial properties of gelatin based biodegradable film over storage of chicken fillet. Results showed that incorporation the both of ZnO and β-glucan have significantly (p<0.01) improved the barrier properties against both of moisture absorption and water vapor permeability and the best properties obtained with film containing 5% of ZnO and 20% β-glucan. In addition, based on microbial tests result, it was obvious that the ZnO loaded films have antimicrobial properties and the highest inhibition activity obtained with 5% of ZnO against the all of studied pathogenic bacteria. Accordingly, the film containing 5% ZnO and 20% β-glucan is introduced as the most effective film for packaging of chicken fillets.     

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Aims: Glucantime has been considered as a drug of choice for treating cutaneous leishmaniasis for many years. In the recent years, resistance to Glucantime has been increasingly reported in some regions of Iran. In the Leishmania, Arsenate/Antimony reductase acts on the basis of thiol metabolism; it can donate the electron from reduced glutaredoxin to pentavalent (sbV) antimony and arsenate and reduce them to trivalent (sbIII) antimony and arsenite, based on its enzymatic property. It has been assumed that a functional mutation in the enzyme can result in drug resistance. In the present study, the role of Sb (V)-As (V) reductase of Leishmania tropica in drug resistant to glucantime was investigated.
Materials and Methods: In the present experimental research, 15 glucantime sensitive samples and 15 glucantime resistant specimens were collected from different regions of Iran through patients with cutaneous leishmaniasis. For mutation detection, first degenerate primers were designed; then, sequencing and simulation techniques were used based on molecular dynamics method.
Findings: In Leishmania tropica-resistant isolates, only one mutation was seen as replacing alanine (Ala) at position 80 instead of valine (Val). The analysis of the radius of gyration did not reveal any increase in the radius of gyration while simulation.
Conclusion: Mutations in glucantime-resistant isolates did not significantly change simulated active site of antimony ion.