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کاربرد نوع شرط:
- جایگاه : پژوهشی
- مجله: Food & Health
- نوع مقاله: Journal Article
- کلمات کلیدی: Superoxide Dismutase,Malondialdehyde,Glutathione peroxidase,Non-Hodgkin lymphoma
- چکیده:
- چکیده انگلیسی: Imbalance in the production of oxidative molecules and antioxidant activity plays an important role in carcinogenesis. This study was performed to evaluate the serum level of malondialdehyde (MDA) as oxidative damage marker, superoxide dismutase (SOD) as well as glutathione peroxidase (GPx) as an antioxidant defense system in non-Hodgkin's lymphoma (NHL). Twenty-five NHL patients and twenty-five healthy individuals were included in the study. The data showed that lower activity of enzymatic antioxidants (GPx, SOD) and higher MDA levels in NHL patients than in the control group. The results suggest that increased serum MDA and decreased SOD and GPx activity may be due to oxidative stress, which may play an important role in NHL formation. the role of oxidative stress in the pathogenesis of the NHL has not been extensively studied. Therefore, the present study aimed to measure the level of MDA as well as GPx and SOD activity in blood tissue collected from NHL patients compared with the control group.
- انتشار مقاله: 03-11-1398
- نویسندگان: Hosnie Hoseini,Parichehreh Yaghmaei,Gholamreza Bahari,Saeed Aminzadeh
- مشاهده
- جایگاه : پژوهشی
- مجله: Iranian Journal of Fisheries Sciences
- نوع مقاله: Journal Article
- کلمات کلیدی: Chitinase,Thermal stability,Serratia marcescens B4A,loop,β chain,unfolding
- چکیده:
- چکیده انگلیسی: Thermostable chitinases are useful for economical, industrial and biotechnological applications. In this paper we attempted to stabilize chitinase from Serratia marcescens B4A by rational mutagenesis and changing of Ser 390 to Ile. This stabilization was performed through entropic stabilization by reduction of the loop length and also by increasing of the beta chain length. On the other hand, with this replacement, polar uncharged residue changed to non-polar one and increased the hydrophobic interactions, furthermore Isoleucine has branched β-carbon that restricted the backbone conformation more than nonbranched residues. Finally all of these factors lead to entropic stabilization and thermal stabilization. The results exhibited that the optimal temperature and pH for enzyme activity of native chitinase weren’t changed by mutagenesis which showed mutation didn’t effect at original characteristics of enzyme, the Km values of native and mutant chitinase was differed very little that showed the affinity of enzyme toward the substrate and natural flexibility of chitinase didn’t changed by mutation, the Vmax value of mutant chitinase was decreased and pH stability of mutant chitinase was increased briefly but its thermal stability was increased remarkably. Mutation made chitinase tolerated high temperatures to 90°C. In addition its activity was increased at 50°C, 60°C for 120 min and up to 2 hours of incubation period and the mutant chitinase showed a high level of activity at 60°C. These results show that entropic stabilization works well for chitinase and this approach may be generally applicable for stabilization of other proteins.
- انتشار مقاله: 16-08-1395
- نویسندگان: Zeinab Emruzi Tubkanlu,Saeed Aminzadeh,Ali Asghar Karkhane,Jahan Alikhajeh,Ahmad Ghoroghi
- مشاهده
- جایگاه : پژوهشی
- مجله: Iranian Journal of Fisheries Sciences
- نوع مقاله: Journal Article
- کلمات کلیدی: Antibiotics,Bacteriocins,Citrus canker,MIC and MBC data,Xanthomonas citri subsp.citri
- چکیده:
- چکیده انگلیسی: Citrus canker is a citrus disease species that created by the bacterium Xanthomonas citri subsp.citri. Many citrus, such as oranges, lime, and grapefruit are affected by the infectious bacteria and stems, leaves and fruits are experiencing loss. In this experiment antibacterial effects of five kinds of antibiotics including ampicillin, kanamycin, chloramphenicol, penicillin, streptomycin, Cronobacteriocin DGH2 and Enterobacteriocin DGH4 were evaluated on 107 strains of Xanthomonas citri subsp.citri. MIC and MBC data for antibiotics and bacteriocins against Xanthomonas strains were performed. According to this project, Xanthomonas strains were comparatively susceptible and resistant to Cronobacteriocin DGH2, Enterobacteriocin DGH4, ampicillin, kanamycin, chloramphenicol, penicillin and streptomycin. NIGEB-183 strain is most sensitive to these antibiotics and bacteriocins, on the other, only NIGEB-242R1 strain is resistant to chloramphenicol. Penicillin is also minimal inhibitory effects on Xanthomonas strains. Based on this case study, chloramphenicol is most antibacterial activity among of antibacterial agents and this agent is a good candidate for inhibitory activity. Cronobacteriocin DGH2 has a moderate antibacterial activity against Xanthomonas strains.
- انتشار مقاله: 03-02-1395
- نویسندگان: Dariush Gholami,Saeed Aminzadeh,Seyed Mehdi Alavi,Nasrin Kazemipour,Ahmad Ghoroghi,Z Emruzi
- مشاهده
- جایگاه : پژوهشی
- مجله: Iranian Journal of Biotechnology
- نوع مقاله: Journal Article
- کلمات کلیدی: Biocontrol,bacteriocin,Protein sequencing,Taguchi orthogonal array,Xanthomonas citri
- چکیده:
- چکیده انگلیسی: Background: Xanthomonas citri subsp. citri (Xcc), the causative agent of bacterial citrus canker, has affected citriculture worldwide. Varieties of means have been used to minimize its devastating effects, but no attention has been given to bacteriocins. Objectives: Here and for the first time, we report the isolation and characterization of two novel bacteriocins. Materials and Methods: Secretome containing bacteriocins of isolated bacteria was separated via SDS-PAGE. Each isolated protein band was characterized and checked for its efficacy in controlling two pathogenic isolates of Xcc via disk diffusion assay. The effects of varieties of carbon, nitrogen and phosphate sources were evaluated on both bacterial growth and bacteriocin production via Taguchi orthogonal method. Results: The two bacteriocins showed an activity up to 55ºC that were sensitive to proteases suggesting being protein in nature. Analysis of SDS-PAGE purified protein bands of bacterial secretomes with demonstrated potency against Xcc revealed the presence of peptides with relative molecular masses of 16.9 and 17 kDa for Cronobacter and Enterobacter, respectively. Sequence analysis of peptides revealed an HCP1 family VI secretion system homologue for Cronobacter (YP_001439956) and pilin FimA homologue for Enterobacter (CBK85798.1). A Taguchi orthogonal array was also implemented to determine the effect of temperature and eight other chemical factors on bacteriocin production for each bacterium. Conclusions: Two peptides with novel antibacterial activities effective against Xcc were isolated, characterized and conditions were optimized for their higher production.
- انتشار مقاله: 17-10-1393
- نویسندگان: Dariush Gholami,Tannaz Goodarzi,Saeed Aminzadeh,Seyed Mehdi Alavi,Nasrin Kazemipour,Naser Farrokhi
- مشاهده
- جایگاه : پژوهشی
- مجله: Iranian Journal of Biotechnology
- نوع مقاله: Journal Article
- کلمات کلیدی: Polymerase Chain Reaction,Site-directed mutagenesis,Nested-Splicing by Overlap Extension PCR
- چکیده:
- چکیده انگلیسی: Background: Splicing by overlap extension (SOE) PCR is used to create mutation in the coding sequence of an enzyme in order to study the role of specific residues in protein’s structure and function. Objectives: We introduced a nested-SOE-PCR (N –SOE-PCR) in order to increase the specificity and generating mutations in a gene by SOE-PCR. Materials and Methods: Genomic DNA from Bacillus thermocatenulatus was extracted. Nested PCR was used to amplify B. thermocatenulatus lipase gene variants, namely wild type and mutant, using gene specific and mutagenic specific primers, followed by cloning in a suitable vector. Briefly in N-SOE-PCR method, instead of two pairs of primers, three pairs of primers are used to amplify a mutagenic fragment. Moreover, the first and second PCR products are slightly longer than PCR products in a conventional SOE. PCR products obtained from the first round of PCR are used for the second PCR by applying the nested and mutated primers. Following to the purification of the amplified fragments, they will be subject of the further purification and will be used as template to perform the third round of PCR using gene specific primers. In the end, the products will be cloned into a suitable vector for subsequent application. Results: In comparison to the conventional SOE-PCR, the improved method (i.e. N-SOE-PCR) increases the yield and specificity of the products. In addition, the proposed method shows a large reduction in the non-specific products. Conclusions: By applying two more primers in the conventional SOE, the specificity of the method will be improved. This would be in part due to annealing of the primers further inside the amplicon that increases both the efficiency and a better attachment of the primers. Positioning of the primer far from both ends of an amplicon leads to an enhanced binding as well as increased affinity in the third round of amplification in SOE.
- انتشار مقاله: 09-09-1393
- نویسندگان: Ali Asghar Karkhane,Bagher Yakhchali,Ferdous Rastgar Jazii,Bijan Bambai,Saeed Aminzadeh,Fatemeh Rahimi
- مشاهده
- جایگاه : پژوهشی
- مجله: Iranian Journal of Biotechnology
- نوع مقاله: Journal Article
- کلمات کلیدی: Wheat,Laying hen,Xylanase,Fat digestibility,Intestinal enzymes
- چکیده:
- چکیده انگلیسی: The effects of non-starch polysaccharide (NSP) content of wheat and xylanase supplementation (XS) of the diet, on intestinal enzyme activity (amylase, aminopeptidase and lipase), fat digestibility and ileal viscosity of laying hens has been studied. Two hundred forty Hy-Line W-36 layer from 20 to 25 week of age were studied under a factorial experiment (4×2) in completely randomized design with 8 treatments including four levels of wheat (0, 23, 46 and 69%) corresponding to a dietary xylose content of 1.9, 2.1, 2.3 and 2.5% and two levels of xylanase (none or added at the dosage recommended by the supplier). Each treatment was replicated five times each with six hens. Wheat inclusion in the diet increased amylase and lipase activity in the duodenum and jejunum, respectively (P<0.001). Wheat inclusion, increased ileal viscosity while adding xylanase to diet, reduced it (P<0.001). Fat digestibility was decreased by wheat increment levels (P<0.001). The pH of the digesta content in different segments of gastro-intestinal tract (GIT) was not affected by wheat levels or XS, except for cecum that decreased with increasing the level of wheat. Relative weight and length of duodenum,
jejunum and ileum were not affected by dietary treatment. Results suggested that wheat inclusion at high
levels increased endogenous enzyme activity but could not alleviate the adverse effect of NSP content of
diet on fat digestibility. Keywords: Fat digestibility; Intestinal enzymes; Laying hen; Wheat; Xylanase- انتشار مقاله: 11-04-1391
- نویسندگان: Sara Mirzaie,Mojtaba Zaghari,Saeed Aminzadeh,Mahmoud Shivazad
- مشاهده
- جایگاه : پژوهشی
- مجله: Iranian Journal of Biotechnology
- نوع مقاله: Journal Article
- کلمات کلیدی: Screening,Taguchi Method,Isolation,Chitinase,Serratia marcescens B4A
- چکیده:
- چکیده انگلیسی: Chitinase production by newly isolated Serratia marcescens B4A was optimized following Taguchi’s
array methods. Twenty-three bacterial isolates were screened from shrimp culture ponds in the South of
Iran. A chitinase-producing bacterium was isolated based on it’s ability to utilize chitin as the sole carbon
source. The isolate designated as B4A, was identified as Serratia marcescens based on its 16S rRNA
sequence and key morphological, physiological and biochemical characteristics. The cultivation of Serratia
marcescens B4A in the appropriate liquid medium resulted in production of high levels of chitinase. The
malt extract and colloidal chitin represented the best nitrogen and carbon sources, respectively. Chitinase
production by Serratia marcescens B4A was optimized following the Taguchi orthogonal array (OA) for the
design of experiments (DOE). Statistical experimental design via the Taguchi method was applied to determine the optimal levels of physical parameters and key media components in the medium, such as temperature, pH, NaCl and chitin concentrations. The results of this study showed that temperature of 30ºC, pH 7.9, NaCl 0.1% (w/v) and chitin 1% (w/v) are optimal conditions for this protocol.
- انتشار مقاله: 09-07-1389
- نویسندگان: Mandana Zarei,Saeed Aminzadeh,Hossein Zolgharnein,Alireza Safahieh,Ahmad Ghoroghi,Abbasali Motallebi,Morteza Daliri,Abbas Sahebghadam Lotfi
- مشاهده
- جایگاه : پژوهشی
- مجله: Food & Health
- نوع مقاله: Journal Article
- کلمات کلیدی: Superoxide Dismutase,Malondialdehyde,Glutathione peroxidase,Non-Hodgkin lymphoma
- چکیده:
- چکیده انگلیسی: Imbalance in the production of oxidative molecules and antioxidant activity plays an important role in carcinogenesis. This study was performed to evaluate the serum level of malondialdehyde (MDA) as oxidative damage marker, superoxide dismutase (SOD) as well as glutathione peroxidase (GPx) as an antioxidant defense system in non-Hodgkin's lymphoma (NHL). Twenty-five NHL patients and twenty-five healthy individuals were included in the study. The data showed that lower activity of enzymatic antioxidants (GPx, SOD) and higher MDA levels in NHL patients than in the control group. The results suggest that increased serum MDA and decreased SOD and GPx activity may be due to oxidative stress, which may play an important role in NHL formation. the role of oxidative stress in the pathogenesis of the NHL has not been extensively studied. Therefore, the present study aimed to measure the level of MDA as well as GPx and SOD activity in blood tissue collected from NHL patients compared with the control group.
- انتشار مقاله: 03-11-1398
- نویسندگان: Hosnie Hoseini,Parichehreh Yaghmaei,Gholamreza Bahari,Saeed Aminzadeh
- مشاهده
- جایگاه : پژوهشی
- مجله: Advanced Research in Microbial Metabolites & Technology
- نوع مقاله: Journal Article
- کلمات کلیدی: Amylopullulanase,Pullulan,Recombinant protein,Zymogram
- چکیده:
- چکیده انگلیسی: Starch debranching enzymes that merely hydrolyze α-(1→6) glycosidic linkages are classified into isoamylases (EC 3.2.1.68) and pullulanases (EC 3.2.1.41). An exception to this definition would be amylopullulanase, a type of pullulanase that is capable of cleaving both α-(1→4) and α-(1→6) linkages. Amylopullulanases are in demand in liquid sugar industries to generate glucose and some other starch derivatives. Pullulanases can be used in conjunction with amylases to improve sugar availability during sugar syrup production. Here, a thermophilic Cohnella sp. A01 amylopullulanase (EC 3.2.1.41) gene, namely Coh4159, was PCR amplified and cloned in pET-26b(+) and transformed into BL21(DE3). Recombinant Coh4159 was heterologously expressed in the presence of 0.5 mM IPTG and purified via affinity chromatography, and further characterized. Enzyme activity was demonstrated via zymogram analysis in the presence of pullulan. The enzyme had a hydrolytic effect on pullulan with Vmax = 2.85 µmol.min-1 and Km = 0.5 mM. Temperature optima and pH were 60 ˚C and 6.0. In which the enzyme kept its activity at wide pH (4-9) and temperature (30-70 ˚C) ranges. The recombinant enzyme kept 50% of its activity for 60 min, 100 min and 120 min when incubated at 80, 70 and 60 ˚C, respectively. Amongst metal ions tested, Mn2+, and Ca2+ have improved the enzyme activity both at 5 and 10 mM. The results promise the capability of producing a commercial industrial enzyme, well-suited to liquid sugar syrup industry specification.
- انتشار مقاله: 02-11-1396
- نویسندگان: Parvin Valiulahi,Saeed Aminzadeh,Mehdi Shamsara,Naser Farrokhi,Jahan Alikhajeh
- مشاهده