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Leptinotarsa decemlineata (Say) and Lasioderma serricorne F. are destructive pre-harvest and post-harvest pests of many plants in the family Solanaceae, and stored foodstuffs and non-food items, respectively. In this study, some biochemical characteristics of cellulase in the larval digestive tract of these pests were studied. Endo-β-1, 4-glucanase activity was measured against the substrate carboxyl methyl cellulose. Maximum activity of the enzyme in L. decemlineata and L. serricorne occurred at pH 7.0 and pH 6.0, respectively. The enzymes from L. decemlineata and L. serricorne were maximally stable at pH 7.0 and pH 5-6, respectively. However, the enzyme extracted from L. serricorne is more stable than that of L. decemlineata. Cellulase activity was in the highest level at 50 °C in both species. EDTA and SDS reduced cellulase activity, while the Ca²⁺, Mg²⁺ and Na⁺ ions had a significant increasing effect on cellulase activity. K⁺ did not have any significant effect on the enzyme activity. The values of Km and Vmax were 0.608 % and 0.0187 µmol min^-1 mg^-1 protein in L. decemlineata, and 0.99 %, and 0.0035 µmol min^-1 mg^-1 protein in L. serricorne, respectively. Zymogram studies revealed two bands of cellulase activity in the digestive tract of both species.  

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This study was undertaken to find out the optimum physicochemical parameters of fermentation, i.e. pH, incubation temperature and incubation time for the cellulase enzyme production of Trichoderma harzianum. The extracellular protein content was estimated by the dye binding method of Bradford. Endo-glucanase (EG), exoglucanase (or Cellobiohydrolase; CBH), β-glucosidase and total cellulase activity were investigated. The molecular weight of cellulase enzymes was studied using SDS-PAGE. To identify the predominant catalytic components in optimum conditions of enzyme production, cellulases were separated by an adapted two-dimensional electrophoresis technique. Estimated optimum conditions for cellulase enzyme were found as: pH 6.5, incubation temperature 28°C and incubation time 72 h. The SDS-PAGE profiles showed several enzyme bonds such as CBHs, EGs and BGLs. The T. harzianumhad both enzyme bonds of Cel7A (CBHI) and Cel7B (EG). Finally, the results of the 2D PAGE analysis showed that the profile of protein in optimium conditions of enzyme production had several enzymes such as CBHs, EGs and the high values of cellulose activity due to synergism that occurred between the CBH and EG.
 

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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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Backgrounds: Streptomyces is an aerobic filamentous Gram-positive bacterium frequently found in various environments worldwide. Cellulases are a group of glycosyl hydrolase enzymes that are frequently produced by bacteria. Thus, the aim of this study was to detect cellulase-encoding gene (celA) in soil-living Streptomyces strains and evaluate its cloning in Escherichia coli Origami strain.
Materials & Methods: Soil samples were collected from a depth of 5-10 cm in Tehran, Iran. After identification of Streptomyces isolates by morphological and biochemical tests, genomic DNA was extracted. Polymerase chain reaction (PCR) test was employed to identify Streptomyces strains harboring the cellulase gene. The celA gene was positively transmitted to the host bacterium E. coli via a vector and cloned through the TA technique. Real-time PCR was used to measure the overexpression of this gene. ClustalX and Mega5 software were used to draw the phylogenetic tree.
Findings: Out of 12 Streptomyces isolates, 25% were found to carry the celA gene. After cloning the celA gene, the cloned strains were chosen by colony selection (blue/white). The real-time PCR test showed the expression of the celA gene in the transformed strains. Phylogenetic analysis results using the neighbor-joining assay showed that Streptomyces spp. with 81% bootstrap were located in the same clade, indicating their close relatedness.
Conclusion: Soil is one of the high-potential sources of the production of secondary metabolites, which could be used as a valuable source of various biological products such as cellulase.

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In this study the cellulytic activity of different species of Iranian Trichoderma isolates including Trichoderma harzianum (NAS-H101), T. aureoviride (NAS-AV106), T. pleuroticola (NAS-P109), T. longibrachiatum (NAS-L110), T. ghanens (NAS-K108), T. virens (NAS- Vi114), T. atroviride (NAS-A113) and T. atroviride (NAS-A112) was studied. The extracellular protein concentration of these isolates was determined by the dye binding method of Bradford. The molecular weight of cellulase enzymes was studied using SDS-PAGE. The lowest extracellular protein production was observed in NAS-K108. The highest Endo and Exo-glucanase activity were observed in NAS-L110 and NAS-A113, respectively. The SDS-PAGE profiles had several enzyme bands such as cellobiohydrolases, endoglucanases and β-glucosidases. The NAS-K108and NAS-P109 had both enzyme bands of CBH I and CBH II, but other isolates had only a sharp enzyme band correlated to CBH I or CBH II. The highest synergy was observed in FPase of NAS-A112, that contained a large amount of Cel 6A (CBH II) and a minor amount of Cel 7B (EG I). The results indicated that NAS-A113 overproduces cellulases, ß-glycosidase, and the extracellular enzymes, which suggest that this species might be utilized as a biological agent and or a source of enzymes for cellulose degradation in colloidal cellulose.
 

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In comparison with traditional extraction methods, aqueous enzymatic extraction of oil from oilseeds is a recent clean technology. This paper reports work performed at laboratory scale to extract soybean oil by aqueous enzymatic extraction method. In the present work the influence of enzymes concentration, extraction time, dilution ratio, particle size and 3 interactions in the final yield are evaluated and process parameters have been optimized by Taguchi method.16 extraction experiments carried out, statistical analysis showed that particle size was the most significant variable in oil extraction. Themaximal oil extraction yield was 61.42%.

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Trichoderma species are known as effective agents used for biological control of plant pathogenic fungi. The Trichoderma harzianum and its mutant isolates were cultured and their traits including, mycelial growth, antagonistic activity and extracellular proteins and enzymes production (Chitinase and Cellulase) were investigated to select the most effective mutant isolates against plant pathogenic fungus Rhizoctonia solani. Also, the purity and composition of enzyme-rich protein samples were evaluated under denaturing gel electrophoresis. This study clearly showed the possibility of improving mycelia growth rate (from 1.18 to 1.33 cm d^-1), the antagonistic capability of Trichoderma (from 54.9% growth inhibition of R. solani to 66%), extracellular proteins and enzymes production for biological control of plant diseases through mutation with γ-radiation. Also, compared to wild type strain, protein production in the mutant isolates increased. Moreover, the highest specific chitinase enzyme activities were observed in mutant isolates T. h M8 (42.48 U mg^-1) and T. h M15 (38.25 U mg^-1). Trichoderma mutant of T. h M8 maintained higher mycelia growth rate and higher ability to inhibit growth of R. solani. The SDS-PAGE profiles had several enzyme protein bands such as CelloBioHydrolases (CBHs), EndoGlucanases (EGs), β-Glucosidases (BGLs), endochitinases, and β-(1, 4)-N-acetyl glucoaminidases. SDS-PAGE analysis indicated the presence of different protein bands in the range of 10.5 to 245 KDa. Interestingly, expression of chitinase in 95 percent of mutants was higher than wild type of T. harzianum. The results showed that gamma mutation could increase the efficiency and amount of enzymes in T. harzianum, while these enzymes are involved in antagonistic properties of T. harzianum.

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Fruit cracking is a predominant physiological disorder of lemon that limits its productivity. The present study aimed to compare the physiological and biochemical traits of cracked and normal fruits of lemon, to understand the cause of fruit cracking and find a viable solution for this disorder. This study was conducted on five-year-old uniform healthy trees grown at fruit research farm, Punjab Agricultural University, Ludhiana, during 2017-2018. Fruits of lemon cracked in different patterns and the cracking peaked due to sudden rainfall and high humidity after a dry spell during the two consecutive years of study. The peel thickness, peel percent and chlorophyll content of the cracked peel was significantly low as compared to the normal ones. Activity of peroxidase and two cell wall degrading enzymes, namely, cellulase and polygalacturonase were higher in cracked peels. Juice content and ascorbic content were low in cracked fruit juice as compared to normal ones. Meanwhile, calcium, potassium and boron content were higher in the normal peel and lower in the cracked peel. A significant positive correlation of fruit cracking incidence with proline, peroxidase, cellulase and polygalacturonase was established, whereas a negative significant correlation was established between fruit cracking percent and peel thickness, calcium, potassium, boron, juice percent and ascorbic acid content. Nutrient deficiency and higher activities of cellulase and polygalacturonase in peel of cracked fruits emerged as the cause of fruit cracking incidence in lemon. Hence, foliar application of calcium, potassium, and boron are recommendable as a remedial measure for prevention of fruit cracking in lemon.