University of Isfahan Journal of Microbial Biology 3060-7647 5 18 2016 08 22 Transformation of Mortierella alpina (fatty acid supplier) myceliums via AMT system (Agrobacterium Mediated Transformation) Transformation of Mortierella alpina (fatty acid supplier) myceliums via AMT system (Agrobacterium Mediated Transformation) 1 10 20379 10.22108/bjm.2016.20379 FA Aida Javanmard M.Sc. of Agricultural biotechnology, Tarbiat Modares University, Tehran, Iran Forough Sanjarian Assistant Professor of Cellular and Molecular biology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran Zohre Hamidy Associate Professor of Biotechnology, Tarbiat Modares University, Tehran, Iran Journal Article 2016 10 04 Introduction: Mortierella alpina is one of the most important fungi in food industry because of having ability of synthesizing unsaturated fatty acids, particularly Arashidonic Acid. This is a precursor of Eicosanoidregulate-lipoprotein metabolism which is involved in blood rheology, platelet activation and leukocyte-function, and the functional characteristics of the cell membrane. <br /> <br />Materials and methods: In this study genetic transformation of M. alpina CBS754.68 fungus was evaluated via Agrobacterium tumefaciens and Agrobacterium rhizogenes. Agrobacteriums containing pBI121 vector were used for transformation of three days of old mycelia. Three days old hyphae were exposed to the bacteria with three level of time (one, two and three hours) in the present of acetosyringone. Mitotic stability of the third generation of transgenic (T2) was confirmed by GUS assay and amplification of CaMV 35S promoter by polymerase chain reaction. <br /> <br />Results: The highest percentage of transformation and mitotic stability were obtained by using A. tumefaciens and A. rhizogenese, respectively. <br /> <br />Discussion and conclusion: The results showed that to obtain more efficient and more stable transformation, the fundamental factor is the use of suitable species of Agrobacterium. It is the first report for transformation of autothroph strain of M. alpine via Agrobacterium. Introduction: Mortierella alpina is one of the most important fungi in food industry because of having ability of synthesizing unsaturated fatty acids, particularly Arashidonic Acid. This is a precursor of Eicosanoidregulate-lipoprotein metabolism which is involved in blood rheology, platelet activation and leukocyte-function, and the functional characteristics of the cell membrane. <br /> <br />Materials and methods: In this study genetic transformation of M. alpina CBS754.68 fungus was evaluated via Agrobacterium tumefaciens and Agrobacterium rhizogenes. Agrobacteriums containing pBI121 vector were used for transformation of three days of old mycelia. Three days old hyphae were exposed to the bacteria with three level of time (one, two and three hours) in the present of acetosyringone. Mitotic stability of the third generation of transgenic (T2) was confirmed by GUS assay and amplification of CaMV 35S promoter by polymerase chain reaction. <br /> <br />Results: The highest percentage of transformation and mitotic stability were obtained by using A. tumefaciens and A. rhizogenese, respectively. <br /> <br />Discussion and conclusion: The results showed that to obtain more efficient and more stable transformation, the fundamental factor is the use of suitable species of Agrobacterium. It is the first report for transformation of autothroph strain of M. alpine via Agrobacterium. https://bjm.ui.ac.ir/article_20379_98f4c883df73fd0bf34926df6706d431.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 5 18 2016 08 22 Cloning of affecting pyruvate decarboxylase gene in the production bioethanol of agricultural waste in the E.coli bacteria Cloning of affecting pyruvate decarboxylase gene in the production bioethanol of agricultural waste in the E.coli bacteria 11 28 20380 10.22108/bjm.2016.20380 FA Masome Zeinali M.Sc. of Agricultural biotechnology, University of Mohaghegh Ardabili, Ardabil, Iran Bahman Hosseini Associate Professor of Plant Biotechnology of Agriculture College and Institute of Biotechnology of Urmia University, Iran Soodabeh Jahanbakhsh Associate Professor of Plant Breeding, University of Mohaghegh Ardabili, Ardabil, Iran Mahmod Rezazadeh Bari Associate Professor of Food Industry of Urmia University, Iran Meisam Tabatabaee Assistant Professor of Microb Biotechnology, Institute of Biotechnology, Karaj, Iran Journal Article 2016 10 04 Introduction: Ethanol made by a biomass is one of the useful strategies in terms of economic and environmental and as a clean and safe energy to replace fossil fuels considered and examined. <br /> <br />Materials and methods: In this study, key enzyme in the production of ethanol (Pyruvate decarboxylase) from Zymomonas mobilis bacteria was isolated and cloned at E. coli bacteria by freeze and thaw method. For gene cloning, we used specific primers of pdc and PCR reaction and then pdc gene isolated and pET 28a plasmid double digested with (Sal I and Xho I) enzymes. Digestion Products were ligated by T4 DNA ligase in 16 °C for 16 hours. <br /> <br />Results: Results of bacteria culture showed that a few colonies containing pET 28a plasmid could grow. Result of colony pcr of pdc gene with specific primers revealed 1700 bp bands in 1% agarose gel electrophoresis. The results of PCR with T7 promotor forward primer and pdc revers primer have proved the accurate direction of integration of pdc gene into plasmid and revealed 1885 bp band. Double digestion of recombinant plasmid with SalI and XhoI enzymes revealed same bands. Finally, RT showed the expected band of 1700 bp that implies the desired gene expression in the samples. <br /> <br />Discussion and conclusion: Due to the increased production of ethanol via pyruvate decarboxylase gene cloning in expression plasmids with a strong promoter upstream of the cloning site can conclude that, pyruvate decarboxylase cloning as a key gene would be useful and according to beneficial properties of E. coli bacteria, transfering the gene to bacteria appears to be reasonable. Introduction: Ethanol made by a biomass is one of the useful strategies in terms of economic and environmental and as a clean and safe energy to replace fossil fuels considered and examined. <br /> <br />Materials and methods: In this study, key enzyme in the production of ethanol (Pyruvate decarboxylase) from Zymomonas mobilis bacteria was isolated and cloned at E. coli bacteria by freeze and thaw method. For gene cloning, we used specific primers of pdc and PCR reaction and then pdc gene isolated and pET 28a plasmid double digested with (Sal I and Xho I) enzymes. Digestion Products were ligated by T4 DNA ligase in 16 °C for 16 hours. <br /> <br />Results: Results of bacteria culture showed that a few colonies containing pET 28a plasmid could grow. Result of colony pcr of pdc gene with specific primers revealed 1700 bp bands in 1% agarose gel electrophoresis. The results of PCR with T7 promotor forward primer and pdc revers primer have proved the accurate direction of integration of pdc gene into plasmid and revealed 1885 bp band. Double digestion of recombinant plasmid with SalI and XhoI enzymes revealed same bands. Finally, RT showed the expected band of 1700 bp that implies the desired gene expression in the samples. <br /> <br />Discussion and conclusion: Due to the increased production of ethanol via pyruvate decarboxylase gene cloning in expression plasmids with a strong promoter upstream of the cloning site can conclude that, pyruvate decarboxylase cloning as a key gene would be useful and according to beneficial properties of E. coli bacteria, transfering the gene to bacteria appears to be reasonable. https://bjm.ui.ac.ir/article_20380_5ff676a727830c635a147ce6f5125499.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 5 18 2016 08 22 Isolation, cloning and analysis of the hexose transporter 6 gene (HXT6) in a native strain of Saccharomyces cerevisiae IBRC-M30069 Isolation, cloning and analysis of the hexose transporter 6 gene (HXT6) in a native strain of Saccharomyces cerevisiae IBRC-M30069 29 40 20381 10.22108/bjm.2016.20381 FA Solmaz Azizi M.Sc. student of Agricultural Biotechnology, Azarbaijan Shahid Madani University, Tabriz, Iran Alireza Tarinejad Associate Professor of Agricultural Biotechnology, Azarbaijan Shahid Madani University, Tabriz, Iran Mohammad Pazhang Associate Professor of cell and molecular biology, Azarbaijan Shahid Madani University, Tabriz, Iran Journal Article 2016 10 04 Introduction: The Saccharomyces cerevisiae yeast is one of the most important microorganisms to produce ethanol. The S. cerevisiae has 20 genes that encode hexose transporter proteins. Among these gene families, seven genes HXT7-HXT1 have important an role in alcohol production. The researchers proved that alcohol fermentation goes up by increasing the expression of these genes which results in increasing ethanol production. <br /> <br />Materials and methods: In this research, isolation of HXT6 gene by specific primers via PCR technology was achieved. The amplified fragments were cloned into pGEM-T vector and transformed to Escherichia coli and sequence analysis was carried out. <br /> <br />Results: The nucleotide sequence of open reading frame of HXT6 gene revealed a 1713 bp long with a deduced amino acid of 570 residues. The estimated molecular mass and the predicted isoelectric point of the deduced polypeptide were 62.68 kDa and 7.89 respectively. <br /> <br />Discussion and conclusion: The deduced protein sequence showed a high similarity to Hxt6p VL31(EGA76254.1) sequences registered in NCBI and also the highest similarity of this gene with HXT7 ( one of the hexose transporter) was observed. This finding shows that this gene (HXT6) and also HXT7 gene resulted from one ancestor gene by mutation in their functional domain during years. Introduction: The Saccharomyces cerevisiae yeast is one of the most important microorganisms to produce ethanol. The S. cerevisiae has 20 genes that encode hexose transporter proteins. Among these gene families, seven genes HXT7-HXT1 have important an role in alcohol production. The researchers proved that alcohol fermentation goes up by increasing the expression of these genes which results in increasing ethanol production. <br /> <br />Materials and methods: In this research, isolation of HXT6 gene by specific primers via PCR technology was achieved. The amplified fragments were cloned into pGEM-T vector and transformed to Escherichia coli and sequence analysis was carried out. <br /> <br />Results: The nucleotide sequence of open reading frame of HXT6 gene revealed a 1713 bp long with a deduced amino acid of 570 residues. The estimated molecular mass and the predicted isoelectric point of the deduced polypeptide were 62.68 kDa and 7.89 respectively. <br /> <br />Discussion and conclusion: The deduced protein sequence showed a high similarity to Hxt6p VL31(EGA76254.1) sequences registered in NCBI and also the highest similarity of this gene with HXT7 ( one of the hexose transporter) was observed. This finding shows that this gene (HXT6) and also HXT7 gene resulted from one ancestor gene by mutation in their functional domain during years. https://bjm.ui.ac.ir/article_20381_bc40b5ecb91a25f4342900480e91524b.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 5 18 2016 08 22 Production of extracellular L- asparaginase by halophilic bacterium vibrio sp. Production of extracellular L- asparaginase by halophilic bacterium vibrio sp. 41 54 20382 10.22108/bjm.2016.20382 FA Mahdis Zolfaghar M.Sc. of Microbial biotechnology, University of Tehran, Iran Mohammad Ali Amoozegar Associate Professor of Microbiology, University of Tehran, Iran Khosro Khajeh Professor of biochemistry, Tarbiat Modares University, Tehran, Iran Journal Article 2016 10 04 Introduction: L-Asparaginase is an anti-neoplastic drug used in lymphoblastic leukemia chemotherapy. Nowadays, this enzyme derived from bacterial sources, mostly L-asparaginase II from Escherichia coli and in lesser amount L-asparaginase of Erwinia sp. has medical utilization. The long-term usage of these agents leads to allergic reactions; therefore new <br />L- asparaginase with new immunological characteristics is required. Halophilic bacteria might contain L-asparaginase with novel immunological properties that can be used in hypersensitive patients. <br /> <br />Materials and methods: In this study, the production of L-asparaginase by 130 halophilic and halotolerant bacteria isolated from saline environments in Iran was screened. Modified M-9 agar medium was used for qualitative analysis. In the selected strain, growth curve plotting, asparaginase activity assay and analysis for the effect of some factors on the production and enzyme activity were done. <br /> <br />Results: 40 strains produced L-asparaginase. Most of L-asparaginase producers were member of genus Halomonas and Marinobacter (21.4%). GBP­x3 is potent bacteria in L-asparaginase production which belonged to Vibrio sp. based on 16S rRNA analysis. Optimized factors for <br />L-asparaginase production were 350C (0.65 IU/ml), pH 8 (0.926 IU/ml) and 2.5% NaCl (0.903 IU/ml). The highest activity of L-asparaginase was in 350C (0.781 IU/ml), pH 8 (0.82 IU/ml) and without NaCl (0.806 IU/ml). <br /> <br />Discussion and conclusion: The aim of this study was to screen the production of <br />L-asparaginase through 130 halophilic and halotolerant bacteria which were isolated from saline environments in Iran. The results of this work indicate that the bacterium, Vibrio sp. GBP­x3 displays to be potent for asparaginase production. Introduction: L-Asparaginase is an anti-neoplastic drug used in lymphoblastic leukemia chemotherapy. Nowadays, this enzyme derived from bacterial sources, mostly L-asparaginase II from Escherichia coli and in lesser amount L-asparaginase of Erwinia sp. has medical utilization. The long-term usage of these agents leads to allergic reactions; therefore new <br />L- asparaginase with new immunological characteristics is required. Halophilic bacteria might contain L-asparaginase with novel immunological properties that can be used in hypersensitive patients. <br /> <br />Materials and methods: In this study, the production of L-asparaginase by 130 halophilic and halotolerant bacteria isolated from saline environments in Iran was screened. Modified M-9 agar medium was used for qualitative analysis. In the selected strain, growth curve plotting, asparaginase activity assay and analysis for the effect of some factors on the production and enzyme activity were done. <br /> <br />Results: 40 strains produced L-asparaginase. Most of L-asparaginase producers were member of genus Halomonas and Marinobacter (21.4%). GBP­x3 is potent bacteria in L-asparaginase production which belonged to Vibrio sp. based on 16S rRNA analysis. Optimized factors for <br />L-asparaginase production were 350C (0.65 IU/ml), pH 8 (0.926 IU/ml) and 2.5% NaCl (0.903 IU/ml). The highest activity of L-asparaginase was in 350C (0.781 IU/ml), pH 8 (0.82 IU/ml) and without NaCl (0.806 IU/ml). <br /> <br />Discussion and conclusion: The aim of this study was to screen the production of <br />L-asparaginase through 130 halophilic and halotolerant bacteria which were isolated from saline environments in Iran. The results of this work indicate that the bacterium, Vibrio sp. GBP­x3 displays to be potent for asparaginase production. https://bjm.ui.ac.ir/article_20382_64c7761076a84f9c7b2dc1157afc2c46.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 5 18 2016 08 22 Isolation and Identification of L-asparaginase producing Erwinia strains which isolated from Potato Farms Isolation and Identification of L-asparaginase producing Erwinia strains which isolated from Potato Farms 55 66 20383 10.22108/bjm.2016.20383 FA Arastoo Badoei-Dalfard Associate Professors of Biochemistry, Shahid Bahonar University of Kerman, Kerman, Iran Zahra Karami Assistant Professors of Biophysics, Shahid Bahonar University of Kerman, Kerman, Iran Narjes Ramezanipour M.Sc of Microbiology, Shahid Bahonar University of Kerman, Kerman, Iran Journal Article 2016 10 04 Introduction: L-Asparaginase can be effectively used for the treatment of lymphoblastic leukemia. The rapid growth of cancer cells are needed for L-asparagine abundant storage. L-asparaginase catalyzes the hydrolysis of L-asparagine into L-aspartic acid and ammonia. The purpose of this study was to isolate and identify the L-asparaginase producing Erwinia strains from the potato farms of Jiroft. <br /> <br />Materials and methods: Pectolytic Erwinia species isolated from crumbling potato in M9 medium. The desired L-asparaginase producing bacteria were isolated based on the color changes. Biochemical-microbial and the plant pathogenicity tests of these strains were also investigated with potato and geranium. The L-asparaginase production and molecular detection of these Erwinia strains were also investigated. <br /> <br />Results: In this study, L-asparaginase producing Erwinia was isolated on the CVP and M9 mediums. The inoculation of Erwinia strains on the potato and geranium plants showed that Er8 and Er11 species have the ability to cause plant pathogenicity. Results showed that the maximum pathogenicity of Er8 and Er11 was observed after 48 and 15 h of inoculation in potato and geranium plants, respectively. 16S rDNA sequencing and phylogenetic analyses exhibited that Er8 and Er11 strains were similar to Erwinia chrysanthemi with 98% homology. <br /> <br />Discussion and conclusion: Because of several applications of the Erwinia L-asparaginase in various fields, isolated Erwinia and their L-asparaginase can be suitable for applied utilization. Introduction: L-Asparaginase can be effectively used for the treatment of lymphoblastic leukemia. The rapid growth of cancer cells are needed for L-asparagine abundant storage. L-asparaginase catalyzes the hydrolysis of L-asparagine into L-aspartic acid and ammonia. The purpose of this study was to isolate and identify the L-asparaginase producing Erwinia strains from the potato farms of Jiroft. <br /> <br />Materials and methods: Pectolytic Erwinia species isolated from crumbling potato in M9 medium. The desired L-asparaginase producing bacteria were isolated based on the color changes. Biochemical-microbial and the plant pathogenicity tests of these strains were also investigated with potato and geranium. The L-asparaginase production and molecular detection of these Erwinia strains were also investigated. <br /> <br />Results: In this study, L-asparaginase producing Erwinia was isolated on the CVP and M9 mediums. The inoculation of Erwinia strains on the potato and geranium plants showed that Er8 and Er11 species have the ability to cause plant pathogenicity. Results showed that the maximum pathogenicity of Er8 and Er11 was observed after 48 and 15 h of inoculation in potato and geranium plants, respectively. 16S rDNA sequencing and phylogenetic analyses exhibited that Er8 and Er11 strains were similar to Erwinia chrysanthemi with 98% homology. <br /> <br />Discussion and conclusion: Because of several applications of the Erwinia L-asparaginase in various fields, isolated Erwinia and their L-asparaginase can be suitable for applied utilization. https://bjm.ui.ac.ir/article_20383_76dd6950da8dcfca7f6960271bbef9d3.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 5 18 2016 08 22 Effect of carbon and nitrogen sources on carotenoids production by native strain of Aurantiochytrium Ch25 Effect of carbon and nitrogen sources on carotenoids production by native strain of Aurantiochytrium Ch25 67 82 20384 10.22108/bjm.2016.20384 FA Mahdiye Esmizade M.Sc. of Agricultural biotechnology, Graduate University of Advanced Technology, Kerman, Iran Shahryar Shakeri Assistant Professor of Microbiology, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran Mahmood Maleki Assistant Professor of Agricultural biotechnology, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran Journal Article 2016 10 04 Introduction: Microorganisms produce carotenoids as a part of their response to environmental stresses. Carotenoids have many applications in human health, such as antioxidant, anti-cancer, light protection activity and as a precursor for hormones. <br /> <br />Materials and methods: In this study, the effect of different carbon and nitrogen sources was evaluated on carotenoids production by native Aurantiochytrium strain. The effects of different carbon and nitrogen sources were studied on biomass and carotenoid production. Then, carotenoids were extracted and analyzed by TLC, spectrophotometry and HPLC methods. <br /> <br />Results: Results showed that glycerol is the best carbon source for production of high carotenoids content. Selected medium contained: glycerol (1.5% v/v), peptone (1g/l), yeast extract (1g/l) and 50% of sea water. Total carotenoids content was 134.8 µg/g CDW in this medium. TLC analysis showed that the extracted carotenoid is included: beta-carotene, astaxanthin monoester, astaxanthin diester and free astaxanthin. The results of HPLC analysis showed presence of astaxanthin, canthaxanthin, echinenone and β-carotene in the carotenoid extract. <br /> <br />Discussion and conclusion: In this research, production of carotenoids was investigated in native strain of Aurantiochytrium and carotenoids profile was included astaxanthin, canthaxanthin, β-carotene and echinenone. Introduction: Microorganisms produce carotenoids as a part of their response to environmental stresses. Carotenoids have many applications in human health, such as antioxidant, anti-cancer, light protection activity and as a precursor for hormones. <br /> <br />Materials and methods: In this study, the effect of different carbon and nitrogen sources was evaluated on carotenoids production by native Aurantiochytrium strain. The effects of different carbon and nitrogen sources were studied on biomass and carotenoid production. Then, carotenoids were extracted and analyzed by TLC, spectrophotometry and HPLC methods. <br /> <br />Results: Results showed that glycerol is the best carbon source for production of high carotenoids content. Selected medium contained: glycerol (1.5% v/v), peptone (1g/l), yeast extract (1g/l) and 50% of sea water. Total carotenoids content was 134.8 µg/g CDW in this medium. TLC analysis showed that the extracted carotenoid is included: beta-carotene, astaxanthin monoester, astaxanthin diester and free astaxanthin. The results of HPLC analysis showed presence of astaxanthin, canthaxanthin, echinenone and β-carotene in the carotenoid extract. <br /> <br />Discussion and conclusion: In this research, production of carotenoids was investigated in native strain of Aurantiochytrium and carotenoids profile was included astaxanthin, canthaxanthin, β-carotene and echinenone. https://bjm.ui.ac.ir/article_20384_b42700f7ccac7c0dad9494b69ad39145.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 5 18 2016 08 22 Isolation and molecular identification of a UV-resistant strain of Dietzia maris and antioxidant activity of pigment Isolation and molecular identification of a UV-resistant strain of Dietzia maris and antioxidant activity of pigment 83 94 20385 10.22108/bjm.2016.20385 FA Narges Zamanian MSc. of Microbiology, University of Isfahan, Iran Zahra Etemadifar Associate Professor of Microbiology, University of Isfahan, Iran Journal Article 2016 10 04 Introduction: The ability of radioresistant bacteria to survive high levels of UV radiation has been linked to their strong DNA repair systems and ability to produce primary and secondary metabolic products. The biosynthesis of pigments provides an opportunity for bacteria to live in radiation-rich environment. Recent radiation-responsive pigments are used commercially as food colorants, anticancer drugs, as well as antibiotics and for cosmetic purposes. <br /> <br />Materials and methods: Soil sample of Omidiyeh city was collected during the spring of 2014 and UV-resistant strain was isolated after primary and secondary screening. Then it was identified by molecular methods (16S rRNA gene sequencing). Antioxidant activity of pigment was evaluated by 2,2 -diphenyl-1-picryl hydrazyl (DPPH) and the reducing power of pigments were analyzed by ferric chloride. <br /> <br />Results: In this present study, new UV-resistant strain NM2 was isolated and by comparison of these 16S rRNA gene sequences to public database using the BLAST, the genus and species of the isolate was identified as Dietzia maris with 99% similarity. Extraction of pigment from isolated strain was carried out by methanol and acetone as solvents. The spectrum is characterized by maximum peak at 473 nm for pigment of NM2 strain. Antioxidant activity and the reducing ability of pigments increased by increasing their concentrations. NM2 strain pigment showed EC50 concentration of 3.30 mg/ml for DPPH free radical scavenging activity, and EC50 concentration of 28.46 µg/ml for reducing power. <br /> <br />Discussion and conclusion: Isolation of natural resources of pigment is very important with high anti-oxidant activity. In the current study, pigment of UV-resistant bacteria demonstrated a strong antioxidant activity in vitro and pigment of these bacteria could play an important role in UV tolerance. Pigment of UV-resistant bacteria may be an appropriate source for antioxidative-related functional foods and the pharmaceutical industry. Introduction: The ability of radioresistant bacteria to survive high levels of UV radiation has been linked to their strong DNA repair systems and ability to produce primary and secondary metabolic products. The biosynthesis of pigments provides an opportunity for bacteria to live in radiation-rich environment. Recent radiation-responsive pigments are used commercially as food colorants, anticancer drugs, as well as antibiotics and for cosmetic purposes. <br /> <br />Materials and methods: Soil sample of Omidiyeh city was collected during the spring of 2014 and UV-resistant strain was isolated after primary and secondary screening. Then it was identified by molecular methods (16S rRNA gene sequencing). Antioxidant activity of pigment was evaluated by 2,2 -diphenyl-1-picryl hydrazyl (DPPH) and the reducing power of pigments were analyzed by ferric chloride. <br /> <br />Results: In this present study, new UV-resistant strain NM2 was isolated and by comparison of these 16S rRNA gene sequences to public database using the BLAST, the genus and species of the isolate was identified as Dietzia maris with 99% similarity. Extraction of pigment from isolated strain was carried out by methanol and acetone as solvents. The spectrum is characterized by maximum peak at 473 nm for pigment of NM2 strain. Antioxidant activity and the reducing ability of pigments increased by increasing their concentrations. NM2 strain pigment showed EC50 concentration of 3.30 mg/ml for DPPH free radical scavenging activity, and EC50 concentration of 28.46 µg/ml for reducing power. <br /> <br />Discussion and conclusion: Isolation of natural resources of pigment is very important with high anti-oxidant activity. In the current study, pigment of UV-resistant bacteria demonstrated a strong antioxidant activity in vitro and pigment of these bacteria could play an important role in UV tolerance. Pigment of UV-resistant bacteria may be an appropriate source for antioxidative-related functional foods and the pharmaceutical industry. https://bjm.ui.ac.ir/article_20385_529d39d525571e1a06fb9f42a7a31417.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 5 18 2016 08 22 Isolation and molecular identification of Lactobacillus brevis from traditional vinegar Isolation and molecular identification of Lactobacillus brevis from traditional vinegar 95 106 20386 10.22108/bjm.2016.20386 FA Zeynab Ebrahimi MSc. of Microbial biotechnology, University of Isfahan, Iran Abolghasem Esmaeili Associate professor of Molecular biology, University of Isfahan, Iran Tooba Sadat Ahmadi MSc. of Microbial biotechnology, University of Isfahan, Iran Hamid Emami PhD student of Microbiology, University of Isfahan, Iran Mohammad Rabbani Associate Professor of Microbiology, University of Isfahan, Iran 0000-0002-4043-9216 Journal Article 2016 10 04 Introduction: Vinegar is a popular condiment in the world that different materials and methods have been used to produce it. In Iran natural vinegar is also prepared mostly in a traditional way by using different fruits such as grapes and apples. Natural vinegar has beneficent properties and because of this, it is recommended to be used by traditional and Islamic medicine. Vinegar contains acetic acid bacteria, lactic acid bacteria and yeast. Acetic acid bacteria and yeasts are involved in the production of vinegar and lactic acid bacteria improve the flavor of vinegar. The aim of this study was isolation and identification of lactic acid bacteria especially Lactobacillus brevis from traditional vinegar. <br /> <br />Materials and methods: After collecting a few traditional vinegars, the vinegar samples cultured for isolation of lactic acid bacteria on MRS broth and agar media contained nystatin as an anti-yeast antibiotic. Then some microbiological tests including catalase, gram staining and fermentation of carbohydrates were performed. Then, they were cultured at different temperatures, pH and different concentrations of salts. Finally, three isolates bacteria with biochemical properties of Lactobacillus brevis were evaluated by16 srDNA gene amplification. <br /> <br />Results: Twelve lactobacilli were isolated from three vinegar samples. All isolated bacteria were catalase-negative and gram-positive. They could be able to grow at pH around 4.5 and 5.6, and at 2, 4 and 5.6% of salt concentrations. Most of the bacteria grew at 15oC, whereas one isolated grew at 45oC. Sequencing and Blast results showed that the three strains are Lactobacillus brevis. <br /> <br />Discussion and conclusion: Lactobacillus brevis and Lactobacillus plantrum were found in traditional vinegars. Although isolation of Lactobacillus plantrum from vinegar was reported previously, as far as we could determine, it is for the first time that we could isolate Lactobacillus brevis from vinegar. Introduction: Vinegar is a popular condiment in the world that different materials and methods have been used to produce it. In Iran natural vinegar is also prepared mostly in a traditional way by using different fruits such as grapes and apples. Natural vinegar has beneficent properties and because of this, it is recommended to be used by traditional and Islamic medicine. Vinegar contains acetic acid bacteria, lactic acid bacteria and yeast. Acetic acid bacteria and yeasts are involved in the production of vinegar and lactic acid bacteria improve the flavor of vinegar. The aim of this study was isolation and identification of lactic acid bacteria especially Lactobacillus brevis from traditional vinegar. <br /> <br />Materials and methods: After collecting a few traditional vinegars, the vinegar samples cultured for isolation of lactic acid bacteria on MRS broth and agar media contained nystatin as an anti-yeast antibiotic. Then some microbiological tests including catalase, gram staining and fermentation of carbohydrates were performed. Then, they were cultured at different temperatures, pH and different concentrations of salts. Finally, three isolates bacteria with biochemical properties of Lactobacillus brevis were evaluated by16 srDNA gene amplification. <br /> <br />Results: Twelve lactobacilli were isolated from three vinegar samples. All isolated bacteria were catalase-negative and gram-positive. They could be able to grow at pH around 4.5 and 5.6, and at 2, 4 and 5.6% of salt concentrations. Most of the bacteria grew at 15oC, whereas one isolated grew at 45oC. Sequencing and Blast results showed that the three strains are Lactobacillus brevis. <br /> <br />Discussion and conclusion: Lactobacillus brevis and Lactobacillus plantrum were found in traditional vinegars. Although isolation of Lactobacillus plantrum from vinegar was reported previously, as far as we could determine, it is for the first time that we could isolate Lactobacillus brevis from vinegar. https://bjm.ui.ac.ir/article_20386_154e9a5245ad3891770e72859197905a.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 5 18 2016 08 22 Biofilm production among Staphylococcus epidermidis strains isolated from healthy people Biofilm production among Staphylococcus epidermidis strains isolated from healthy people 107 116 20387 10.22108/bjm.2016.20387 FA Fateh Rahimi Assistant Professor of Bacteriology, University of Isfahan, Iran Mohammad Reza Arabestani Assistant Professor of Bacteriology, Hamadan University of Medical Sciences, Iran Journal Article 2016 10 04 Introduction: Staphylococcus epidermidis is well documented as a nosocomial pathogen causing biofilm in patients and healthy people. The aim of this study was to analyze the biofilm formation of S. epidermidis strains isolated from healthy people during 2013-2014. <br /> <br />Materials and methods: Totally 200 healthy people were selected and the sampling was carried out from arm, armpit and axillary area using sterile swaps. Swaps were transferred to thiogylcollate broth and then cultured on mannitol salt agar plates. Isolates were identified at the species level using biochemical tests. Potential of biofilm formation of strains was measured using congo red agar plate and microtiter plate tests. Genes involved in biofilm formation, icaA and icaD, were detected using PCR. <br /> <br />Results: Totally 104 S. epidermidis strains were isolated from healthy people. Amongst these, 66 (63%) and 38 (37%) strains were positive and negative for biofilm formation, respectively. icaA and icaD genes were detected in 100% of strains. <br /> <br />Discussion and conclusion: Prevalence of biofilm producing S. epidermidis isolates among healthy people indicating their colonization with hospital strains. Prevalence of such strains is  urgent for public health. Introduction: Staphylococcus epidermidis is well documented as a nosocomial pathogen causing biofilm in patients and healthy people. The aim of this study was to analyze the biofilm formation of S. epidermidis strains isolated from healthy people during 2013-2014. <br /> <br />Materials and methods: Totally 200 healthy people were selected and the sampling was carried out from arm, armpit and axillary area using sterile swaps. Swaps were transferred to thiogylcollate broth and then cultured on mannitol salt agar plates. Isolates were identified at the species level using biochemical tests. Potential of biofilm formation of strains was measured using congo red agar plate and microtiter plate tests. Genes involved in biofilm formation, icaA and icaD, were detected using PCR. <br /> <br />Results: Totally 104 S. epidermidis strains were isolated from healthy people. Amongst these, 66 (63%) and 38 (37%) strains were positive and negative for biofilm formation, respectively. icaA and icaD genes were detected in 100% of strains. <br /> <br />Discussion and conclusion: Prevalence of biofilm producing S. epidermidis isolates among healthy people indicating their colonization with hospital strains. Prevalence of such strains is  urgent for public health. https://bjm.ui.ac.ir/article_20387_ed08feba968d63a66a7c4e1dfefd87db.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 5 18 2016 08 22 Isolation and Identification of Phototrophic Microorganisms from Rudkhan Castle as a Biodeteriorating Agent Isolation and Identification of Phototrophic Microorganisms from Rudkhan Castle as a Biodeteriorating Agent 117 128 20419 10.22108/bjm.2016.20419 FA Parisa Mohammadi Associate Professor of Microbiology, Alzahra University, Tehran, Iran Paria Gholaminejad MSc. of Microbiology, Alzahra University, Tehran, Iran Marzieh Matinfar MSc. of Microbiology, Alzahra University, Tehran, Iran Mahnaz Gholipour Shahraki MSc. of Microbiology, Alzahra University, Tehran, Iran Ezat Asgarani Associate Professor of Genetics, Alzahra University, Tehran, Iran Journal Article 2014 08 09 <strong>Introduction:</strong>Phototrophic microorganisms are the first residents of different surfaces of ancient buildings’ walls. These organisms can expand to provide the colonization of other microorganisms and to form microbial biofilms. During biofilm growth, acids and metabolites production bore the substratum and cause surfaces damages. Rudkan Castle, the historic monument located in Gilan province, an area with a humid climate has an appropriate surfaces to grow these microorganisms. <br /><strong>Materials and methods:</strong> In this study, morphological identification of algae and cyanobacteria which was isolated from different areas of the brick walls of Castle has been investigated. Samples were taken from 24 different areas of Castle walls and were aseptically cultured into Blue Green Medium (BGM) and Bolds Basal Medium (BBM), and colonies were observed using light microscopy. <br /><strong>Results:</strong> The dominant organisms which were isolated and identified, were <em>Cladophora</em>, <em>Trentepohlia</em>, <em>Klebsormidium</em>, <em>Trebouxia</em>, <em>Pleurastrum</em>, <em>Chlorococcum</em> as chlorophyta order and <em>Scytonema</em>, <em>Tolypothrix</em>, <em>Leptolyngbya</em>, <em>Plectolyngbya</em>, <em>Phormidium</em>, <em>Gloeocapsa</em>, <em>Microcoleus</em>, <em>Nostoc</em> as a cyanobacteria order. <br /><strong>Discussion and conclusion:</strong> As expected, diversity of a large number of cyanobacteria and algae phototrophs was isolated. Diversity of organisms on the surface of monuments depends on many factors, including the weather condition such as moisture, the amount of radiation absorbed by the surface, temperature of area, the materials used in the monuments, the biological acceptance of surface and presence of variety of organisms in the soil surface and the air on. Study of destructive factors is the first step toward the protection and restoration of ancient monuments. <strong>Introduction:</strong>Phototrophic microorganisms are the first residents of different surfaces of ancient buildings’ walls. These organisms can expand to provide the colonization of other microorganisms and to form microbial biofilms. During biofilm growth, acids and metabolites production bore the substratum and cause surfaces damages. Rudkan Castle, the historic monument located in Gilan province, an area with a humid climate has an appropriate surfaces to grow these microorganisms. <br /><strong>Materials and methods:</strong> In this study, morphological identification of algae and cyanobacteria which was isolated from different areas of the brick walls of Castle has been investigated. Samples were taken from 24 different areas of Castle walls and were aseptically cultured into Blue Green Medium (BGM) and Bolds Basal Medium (BBM), and colonies were observed using light microscopy. <br /><strong>Results:</strong> The dominant organisms which were isolated and identified, were <em>Cladophora</em>, <em>Trentepohlia</em>, <em>Klebsormidium</em>, <em>Trebouxia</em>, <em>Pleurastrum</em>, <em>Chlorococcum</em> as chlorophyta order and <em>Scytonema</em>, <em>Tolypothrix</em>, <em>Leptolyngbya</em>, <em>Plectolyngbya</em>, <em>Phormidium</em>, <em>Gloeocapsa</em>, <em>Microcoleus</em>, <em>Nostoc</em> as a cyanobacteria order. <br /><strong>Discussion and conclusion:</strong> As expected, diversity of a large number of cyanobacteria and algae phototrophs was isolated. Diversity of organisms on the surface of monuments depends on many factors, including the weather condition such as moisture, the amount of radiation absorbed by the surface, temperature of area, the materials used in the monuments, the biological acceptance of surface and presence of variety of organisms in the soil surface and the air on. Study of destructive factors is the first step toward the protection and restoration of ancient monuments. https://bjm.ui.ac.ir/article_20419_ebf27c3d9c6c8d9de5a234a81983be2f.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 5 18 2016 08 22 Selection of optimal conditions for anti-corrosive microbial biopolymer production by the Flavobacterium strain using response surface methodology (RSM) Selection of optimal conditions for anti-corrosive microbial biopolymer production by the Flavobacterium strain using response surface methodology (RSM) 129 140 20388 10.22108/bjm.2016.20388 FA Mojtaba Khani MSc. of Chemical Engineering-biotechnology, Institute of Science Biotechnology Malek Ashtar University of Technology, Tehran, Iran Ali Bahrami PhD of Chemical Engineering-biotechnology, Institute of Science Biotechnology Malek Ashtar University of Technology, Tehran, Iran Mohammad Davod Ghafari MSc. of Microbiology, Shahed University, Tehran, Iran Journal Article 2016 10 04 Introduction: Various methods have been proposed to deal with corrosion. One of these methods is using of paints and coatings. In formulation of paints and coatings several anti-corrosion compounds are applied that slow down the corrosion process. In this respect, using microbial biopolymers can improve this problem in the industry with lower costs because of biopolymer production not required to factory and advanced industry. in this study, the effects of temperature, pH and agitation on the biopolymer production using response surface methodology (RSM) were evaluated. <br /> <br />Materials and methods: To produce biopolymer, the culture medium (300 ml) were added in the 500 ml erlenmeyer flasks. Then, the bacterial preculture medium (6% V/V) were inoculated in the flasks and incubated for 96hr in different conditions (agitation speed, tempreture and pH). Afterwards, the medium was centrifuged at 9000 rpm for 10 min and the supernatant was mixed with triple volume of chilled absolute ethanol and stored at 4°C for 24hr to precipitate. <br /> <br />Results: Analysis of the results of design experiments indicate that the biopolymer production­ was strongly governed by the temperature, pH and agitation. The biopolymer production increased steadily up to pH 8 and decreased in the higher pH values. Also, for cell growth suitable temperature was 33°C and maximum concentration of the biopolymer production was agitation of 210 rpm. Finally, maximum concentration of the biopolymer production (14.3g/l) was determined to be in pH of 8, temperature of 33°C and agitation of 210­rpm. <br /> <br />Discussion and conclusion: Anti-corrosive biopolymer production by Flavobacterium sp. affected significantly by physical parameters. The results of the biopolymer production by investigating the conditions of temperature, pH and agitation after optimization, indicates the importance of this parameter for economic production of biopolymer. Introduction: Various methods have been proposed to deal with corrosion. One of these methods is using of paints and coatings. In formulation of paints and coatings several anti-corrosion compounds are applied that slow down the corrosion process. In this respect, using microbial biopolymers can improve this problem in the industry with lower costs because of biopolymer production not required to factory and advanced industry. in this study, the effects of temperature, pH and agitation on the biopolymer production using response surface methodology (RSM) were evaluated. <br /> <br />Materials and methods: To produce biopolymer, the culture medium (300 ml) were added in the 500 ml erlenmeyer flasks. Then, the bacterial preculture medium (6% V/V) were inoculated in the flasks and incubated for 96hr in different conditions (agitation speed, tempreture and pH). Afterwards, the medium was centrifuged at 9000 rpm for 10 min and the supernatant was mixed with triple volume of chilled absolute ethanol and stored at 4°C for 24hr to precipitate. <br /> <br />Results: Analysis of the results of design experiments indicate that the biopolymer production­ was strongly governed by the temperature, pH and agitation. The biopolymer production increased steadily up to pH 8 and decreased in the higher pH values. Also, for cell growth suitable temperature was 33°C and maximum concentration of the biopolymer production was agitation of 210 rpm. Finally, maximum concentration of the biopolymer production (14.3g/l) was determined to be in pH of 8, temperature of 33°C and agitation of 210­rpm. <br /> <br />Discussion and conclusion: Anti-corrosive biopolymer production by Flavobacterium sp. affected significantly by physical parameters. The results of the biopolymer production by investigating the conditions of temperature, pH and agitation after optimization, indicates the importance of this parameter for economic production of biopolymer. https://bjm.ui.ac.ir/article_20388_ae64f429aa4a9f17e07ae3259423de91.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 5 18 2016 08 22 Microbial deproteinization of shrimp shell penaeus merguiensis for chitin extraction Microbial deproteinization of shrimp shell penaeus merguiensis for chitin extraction 141 152 20389 10.22108/bjm.2016.20389 FA Fatemeh Sedaghat MSc. student of Marine biology, Hormozgan University, Iran Morteza Yousefzadi Associate Professor of Biolgogy, University of Hormozgan, Iran Hojjat Toiserkani Associate Professor of Chemistry, University of Hormozgan, Iran Sohrab Najafipour Assistant Professor of Microbiolgogy, Fasa University of Medical Sciences, Iran Journal Article 2016 10 04 Introduction: After cellulose, Chitin is the most abundant biopolymer in nature. The most important derivative of chitin is chitosan, obtained by deacetylation of chitin. Major sources of chitin are the exoskeleton of marine crustaceans such as crab, shrimp, and krill. Chitin extraction from shrimp shells can be carried out chemically or using biological methods. Microbial fermentation as an eco-friendly procedure is a suitable alternative for the chemical and enzymatic processes. In this study, the effect of three protease-producing bacteria species (Pseudomonas aeruginosa, Serratia marcescens, and Bacillus pumilus) on the efficiency of microbial demineralization (DM) and deproteinization (DP) of the shrimp shell penaeus merguiensis, was investigated. Furthermore, the antioxidant activity of hydrolysate obtained during the fermentation process was measured. <br /> <br />Materials and methods: Demineralization and deproteinization was carried out by incubating shrimp waste inoculated with bacteria at 30°C and 100 rpm for 6 days. <br /> <br />Results: Statistical analysis of data showed a significant difference between the percentage of demineralization and deproteinization in different bacteria species (p<0.05). The highest deproteinization (74.76%) and demineralization rate (78.46%) were obtained with P. aeruginosa, while the lowest was observed for S. marcescens. Antioxidant activity of hydrolysate also showed a significant difference. The highest reducing power and total antioxidant capacity were observed in volumes of 400 µl hydrolysate of S.marcescens and 100 µl hydrolysate of B. pumilus, respectively. <br /> <br />Discussion and conclusion: The results indicated that P. aeruginosa in comparison with other bacterial strains, had a higher ability to remove proteins and minerals from shrimp shell waste. Therefore, the use of this bacterium is suitable for protein and minerals removal from marine crustaceans. Introduction: After cellulose, Chitin is the most abundant biopolymer in nature. The most important derivative of chitin is chitosan, obtained by deacetylation of chitin. Major sources of chitin are the exoskeleton of marine crustaceans such as crab, shrimp, and krill. Chitin extraction from shrimp shells can be carried out chemically or using biological methods. Microbial fermentation as an eco-friendly procedure is a suitable alternative for the chemical and enzymatic processes. In this study, the effect of three protease-producing bacteria species (Pseudomonas aeruginosa, Serratia marcescens, and Bacillus pumilus) on the efficiency of microbial demineralization (DM) and deproteinization (DP) of the shrimp shell penaeus merguiensis, was investigated. Furthermore, the antioxidant activity of hydrolysate obtained during the fermentation process was measured. <br /> <br />Materials and methods: Demineralization and deproteinization was carried out by incubating shrimp waste inoculated with bacteria at 30°C and 100 rpm for 6 days. <br /> <br />Results: Statistical analysis of data showed a significant difference between the percentage of demineralization and deproteinization in different bacteria species (p<0.05). The highest deproteinization (74.76%) and demineralization rate (78.46%) were obtained with P. aeruginosa, while the lowest was observed for S. marcescens. Antioxidant activity of hydrolysate also showed a significant difference. The highest reducing power and total antioxidant capacity were observed in volumes of 400 µl hydrolysate of S.marcescens and 100 µl hydrolysate of B. pumilus, respectively. <br /> <br />Discussion and conclusion: The results indicated that P. aeruginosa in comparison with other bacterial strains, had a higher ability to remove proteins and minerals from shrimp shell waste. Therefore, the use of this bacterium is suitable for protein and minerals removal from marine crustaceans. https://bjm.ui.ac.ir/article_20389_81342b3248811d67114ed15a7cd7342e.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 5 18 2016 08 22 Optimization of biotechnological production of xylitol by a Candida tropicalis strain using Response Surface Methodology Optimization of biotechnological production of xylitol by a Candida tropicalis strain using Response Surface Methodology 153 168 20390 10.22108/bjm.2016.20390 FA Omid Zahed MSc. of Food Science, University of Tehran, Karaj, Iran Gholamreza Salehi Jouzani Aossociate Professor of Biotechnology, Agricultural Biotechnology Research Institute of Iran (ABRII), Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran Faramarz Khodayian Associate professor of Food Science, University of Tehran, Karaj, Iran Journal Article 2016 10 04 Introduction: Xylitol is known as one of the most commonly used dietary sugars in food and pharmaceutical industries. The common methodology for production of xylitol is a chemical process which requires high energy and is costly. Biotechnological production of xylitol using microorganisms is an alternative process that is environmentally friendly and cost effective. So, the objective of the present study was to optimize biotechnological production of xylitol from xylose using Candida tropicalis NCIM 3119 strain in the framework of Central Composite Design (CCD) and Response Surface Methodology (RSM). <br /> <br />Materials and methods: Four independent factors including temperature (27, 32 and 37°C), pH (3, 5 and 7), xylose concentration (30, 50 and 70 g/l) and yeast extract concentration (3, 7.5 and 12 g/l) were selected, and the xylitol yield (Yp/s= gram xylitol per gram xylose utilized) and biomass production were calculated. <br /> <br />Results: Based on the constructed model, maximum expected xylitol yield (Yp/s= 0.73) was achieved when temperature, pH, and xylose and yeast extract concentrations were 32.7°C, 4.7, 54.2 g/l and 12 g/l, respectively. To confirm the calculated model, an experiment for xylitol production by the strain in the optimum condition was designed at Erlenmeyer level. The results showed that observed xylitol yield and concentration and also biomass of the strain were 0.69, 36.7 g/l and 11.1 g/l, respectively, which were in accordance with the model. <br /> <br />Discussion and conclusion: Based on the results, it could be concluded that the environmental parameters, including nitrogen source, temperature, pH and xylose and nitrogen source concentrations were optimized to enhance biotechnological production of xylitol, and the final concentration of 36.7 g/l xylitol with 0.69 yield efficiency was achieved. Introduction: Xylitol is known as one of the most commonly used dietary sugars in food and pharmaceutical industries. The common methodology for production of xylitol is a chemical process which requires high energy and is costly. Biotechnological production of xylitol using microorganisms is an alternative process that is environmentally friendly and cost effective. So, the objective of the present study was to optimize biotechnological production of xylitol from xylose using Candida tropicalis NCIM 3119 strain in the framework of Central Composite Design (CCD) and Response Surface Methodology (RSM). <br /> <br />Materials and methods: Four independent factors including temperature (27, 32 and 37°C), pH (3, 5 and 7), xylose concentration (30, 50 and 70 g/l) and yeast extract concentration (3, 7.5 and 12 g/l) were selected, and the xylitol yield (Yp/s= gram xylitol per gram xylose utilized) and biomass production were calculated. <br /> <br />Results: Based on the constructed model, maximum expected xylitol yield (Yp/s= 0.73) was achieved when temperature, pH, and xylose and yeast extract concentrations were 32.7°C, 4.7, 54.2 g/l and 12 g/l, respectively. To confirm the calculated model, an experiment for xylitol production by the strain in the optimum condition was designed at Erlenmeyer level. The results showed that observed xylitol yield and concentration and also biomass of the strain were 0.69, 36.7 g/l and 11.1 g/l, respectively, which were in accordance with the model. <br /> <br />Discussion and conclusion: Based on the results, it could be concluded that the environmental parameters, including nitrogen source, temperature, pH and xylose and nitrogen source concentrations were optimized to enhance biotechnological production of xylitol, and the final concentration of 36.7 g/l xylitol with 0.69 yield efficiency was achieved. https://bjm.ui.ac.ir/article_20390_7f2725f2b0177bf6757f8419ed03b539.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 5 18 2016 08 22 Effect of metal ions on the activity and stability of wild type and mutant pyrazinamidases Effect of metal ions on the activity and stability of wild type and mutant pyrazinamidases 169 182 20391 10.22108/bjm.2016.20391 FA Mehrnoosh Safarzadeh M.Sc. of Cellular and Molecular biology, Azarbaijan Shahid Madani University, Tabriz, Iran Mohammad Pazhang Assistant Professor of Biochemistry, Azarbaijan Shahid Madani University, Tabriz, Iran Framarz Mehrnejad Assistant Professor of Biophysics, University of Tehran, Tehran, Iran Farahnoosh Dustdar Assistant Professor of Microbiology, Shahid Beheshti University of Medical Sciences, Tehran, Iran Nader Chaparzadeh Associate Professor of Plant physiology, Azarbaijan Shahid Madani University, Tabriz, Iran Davoud Rabiei Faradonbeh M.Sc. of Cellular and Molecular biology, Azarbaijan Shahid Madani University, Tabriz, Iran Journal Article 2016 10 04 Introduction: Pyrazinamidase is a metalloenzyme with hydrolyzing activity which is responsible for conversion of pyrazinamide (anti tuberculosis drug) to active molecule, pyrazinoic acid. The metal-binding site in the enzyme is composed of Asp49, His51, His56 and His71. Mutations in the pyrazinamidase gene are responsible for resistance to pyrazinamide in Mycobacterium tuberculosis and can alter the binding of metal ions to the metal binding site in the enzyme. Therefore, it is important to study the effect of metal ions on the enzymatic activity and stability of the wild type and mutant pyrazinamidases. <br /> <br />Materials and methods: In this study, E. coli BL21 was transformed by expression vectors carrying wild type and mutant pyrazinamidase genes. The recombinant proteins were expressed and then purified by Ni- agarose column. The purity of the purified proteins was analyzed by SDS-PAGE electrophoresis. Finally, the activity and stability of the purified enzymes were studied in the presence of 1mM of Ni2+, Fe2+ and Mn2+. <br /> <br />Results: SDS-PAGE analysis showed that the expressed enzymes were purified. The activity and stability results depicted that Ni2+ increases the activity and stability of the wild type and mutant (mutant1: L151S, and mutant2 (triple): A143T/T168A/E173K) enzymes. Fe2+ decreased the activity and stability of mutant1, but has no significant effect on other enzymes. The activity and stability of the enzymes decreased in the presence of Mn2+. <br /> <br />Discussion and conclusion: Ni2+ interact favorably with metal binding site of the wild type enzyme and mutants compared with Fe2+ and Mn2+ and then increases the activity and stability of the enzymes. Introduction: Pyrazinamidase is a metalloenzyme with hydrolyzing activity which is responsible for conversion of pyrazinamide (anti tuberculosis drug) to active molecule, pyrazinoic acid. The metal-binding site in the enzyme is composed of Asp49, His51, His56 and His71. Mutations in the pyrazinamidase gene are responsible for resistance to pyrazinamide in Mycobacterium tuberculosis and can alter the binding of metal ions to the metal binding site in the enzyme. Therefore, it is important to study the effect of metal ions on the enzymatic activity and stability of the wild type and mutant pyrazinamidases. <br /> <br />Materials and methods: In this study, E. coli BL21 was transformed by expression vectors carrying wild type and mutant pyrazinamidase genes. The recombinant proteins were expressed and then purified by Ni- agarose column. The purity of the purified proteins was analyzed by SDS-PAGE electrophoresis. Finally, the activity and stability of the purified enzymes were studied in the presence of 1mM of Ni2+, Fe2+ and Mn2+. <br /> <br />Results: SDS-PAGE analysis showed that the expressed enzymes were purified. The activity and stability results depicted that Ni2+ increases the activity and stability of the wild type and mutant (mutant1: L151S, and mutant2 (triple): A143T/T168A/E173K) enzymes. Fe2+ decreased the activity and stability of mutant1, but has no significant effect on other enzymes. The activity and stability of the enzymes decreased in the presence of Mn2+. <br /> <br />Discussion and conclusion: Ni2+ interact favorably with metal binding site of the wild type enzyme and mutants compared with Fe2+ and Mn2+ and then increases the activity and stability of the enzymes. https://bjm.ui.ac.ir/article_20391_56c8aeacdb84ab0bbd0a563b2d3ba2d2.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 5 18 2016 08 22 The potato dry rot causal agent and severity in Ardabil storages and the resistance of cultivars to the disease The potato dry rot causal agent and severity in Ardabil storages and the resistance of cultivars to the disease 183 202 20392 10.22108/bjm.2016.20392 FA Leila Khoshnevis MSc. Student of Plant Pathology, University of Guilan, Iran Ahmad Rouhibakhsh Assistant Professor of Plant Virology, University of Guilan, Iran Sedigheh Mousanejad Assistant Professor of Plant Pathology, University of Guilan, Iran Journal Article 2016 10 04 Introduction: Dry rot is one of the most important diseases of potato in storages. The aim of this study was to determine dry rot severity in potato storages of Ardabil, identify the disease causal agents and evaluate some potato cultivars resistance to the disease. <br /> <br />Materials and methods: Totally 150 infected samples were collected from thirty nine studied storages in Ardabil. Dry rot severity and the prevalence of infected tubers were determined by surveying three 50 kg bags of potato in the storages. Fungi isolated and purified from the tubers with dry rot symptoms. A factorial design with four replications was applied in order to evaluate the reaction of five potato cultivars to four Fusarium species and determining the resistant one to dry rot. Potato tuber slices were inoculated by conidial suspension of Fusarium species. Four days after inoculation and maintaining in darkness and 25oC, the cultivars susceptibility was  identified. <br /> <br />Results: Four species of Fusarium were identified as dry rot causal agents in Ardabil as follows: F. oxysporum, F. poae, F. solani and F. sporotrichioides. There was no significant difference between Fusarium species in pathogenicity. The reaction of cultivars to various Fusarium species was different. Boren had the lowest infection (12.5%) and Sabalan showed the highest (98.87 %). Sabalan ،Khavaran ،Agria and hybrid line 3970093 displayed the highest susceptibility to various studied Fusarium species. <br /> <br />Discussion and conclusion: Based on the results, Boren is introduced as the most resistant cultivar to Fusarium dry rot and Sabalan, Agria and hybrid line 3970093 as the susceptible ones. Introduction: Dry rot is one of the most important diseases of potato in storages. The aim of this study was to determine dry rot severity in potato storages of Ardabil, identify the disease causal agents and evaluate some potato cultivars resistance to the disease. <br /> <br />Materials and methods: Totally 150 infected samples were collected from thirty nine studied storages in Ardabil. Dry rot severity and the prevalence of infected tubers were determined by surveying three 50 kg bags of potato in the storages. Fungi isolated and purified from the tubers with dry rot symptoms. A factorial design with four replications was applied in order to evaluate the reaction of five potato cultivars to four Fusarium species and determining the resistant one to dry rot. Potato tuber slices were inoculated by conidial suspension of Fusarium species. Four days after inoculation and maintaining in darkness and 25oC, the cultivars susceptibility was  identified. <br /> <br />Results: Four species of Fusarium were identified as dry rot causal agents in Ardabil as follows: F. oxysporum, F. poae, F. solani and F. sporotrichioides. There was no significant difference between Fusarium species in pathogenicity. The reaction of cultivars to various Fusarium species was different. Boren had the lowest infection (12.5%) and Sabalan showed the highest (98.87 %). Sabalan ،Khavaran ،Agria and hybrid line 3970093 displayed the highest susceptibility to various studied Fusarium species. <br /> <br />Discussion and conclusion: Based on the results, Boren is introduced as the most resistant cultivar to Fusarium dry rot and Sabalan, Agria and hybrid line 3970093 as the susceptible ones. https://bjm.ui.ac.ir/article_20392_443ffac14906d2ff63371db9b8d9d5e1.pdf