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Journal of Molecular Microbiology and Biotechnology

Ashraf Mohabati Mobarez, Roya Ahmadrajabi, Nima Khoramabadi, Ali-Hatef Salmanian
To investigate the immunoprotective effects of the recombinant type A flagellin (FLA), the flaA gene of Legionella pneumophila serogroup 1 strain Paris was cloned into pET28a(+). Recombinant FLA (rFLA) was overexpressed in E. coli BL21 (DE3) and purified by Ni2+ exchange chromatography. Female BALB/c aged 6-8 weeks were immunized with 20 μg of rFLA. Nonimmunized mice along with mice inoculated with a sublethal dose of live L. pneumophila intravenously were considered as negative and positive controls, respectively...
April 26, 2017: Journal of Molecular Microbiology and Biotechnology
Yao Zhai, Zhao Zhang, Zhanwei Wang, Yusheng Chen, Qi Wang, Yuan Lv, Jingping Yang, Tong Zhao, Yatao Guo, Zhancheng Gao
AIMS: This work investigated the relative strengths of different blaSHV promoter-associated sequences and their regulation function in blaSHV expression and β-lactam resistance. METHODS: Recombinant plasmids with the promoter-associated sequences (P-W, P-S, P-IS, and P-WPD), tac promoter, and combined fragments of promoter and blaSHV were separately constructed and transformed into Escherichia coli DH5α. The relative strengths of the promoters indicated by the intensities of green fluorescent protein and the mRNA expression levels of blaSHV were compared...
March 23, 2017: Journal of Molecular Microbiology and Biotechnology
Jolanta Mierzejewska, Aleksandra Tymoszewska, Karolina Chreptowicz, Kamil Krol
2-Phenylethanol (2-PE) is an aromatic alcohol with a rosy scent which is widely used in the food, fragrance, and cosmetic industries. Promising sources of natural 2-PE are microorganisms, especially yeasts, which can produce 2-PE by biosynthesis and biotransformation. Thus, the first challenging goal in the development of biotechnological production of 2-PE is searching for highly productive yeast strains. In the present work, 5 laboratory Saccharomyces cerevisiae strains were tested for the production of 2-PE...
February 24, 2017: Journal of Molecular Microbiology and Biotechnology
Hanna S Ruppersberg, Maren R Goebel, Svea I Kleinert, Daniel Wünsch, Kathleen Trautwein, Ralf Rabus
To more efficiently process the large sample numbers for quantitative determination of ammonium (NH4+) and phosphate (orthophosphate, PO43-) generated during comprehensive growth experiments with the marine Roseobacter group member Phaeobacter inhibens DSM 17395, specific colorimetric assays employing a microplate reader (MPR) were established. The NH4+ assay is based on the reaction of NH4+ with hypochlorite and salicylate, yielding a limit of detection of 14 µM, a limit of quantitation of 36 µM, and a linear range for quantitative determination up to 200 µM...
February 8, 2017: Journal of Molecular Microbiology and Biotechnology
Nina El Najjar, Christine Kaimer, Thomas Rösch, Peter L Graumann
Bacillus subtilis possesses 2 DNA translocases that affect late stages of chromosome segregation: SftA separates nonsegregated DNA prior to septum closure, while SpoIIIE rescues septum-entrapped DNA. We provide evidence that SftA is associated with the division machinery via a stretch of 47 amino acids within its N-terminus, suggesting that SftA is recruited by protein-protein interactions with a component of the division machinery. SftA was also recruited to mid-cell in the absence of its first 20 amino acids, which are proposed to contain a membrane-binding motif...
2017: Journal of Molecular Microbiology and Biotechnology
Huayou Chen, Zhi Chen, Bangguo Wu, Jawad Ullah, Tianxi Zhang, Jinru Jia, Hongcheng Wang, Tianwei Tan
In the present study, fusion genes composed of Thermotoga maritima MSB8 nitrilase and Bacillus subtilis 168 outer coat protein CotG were constructed with various peptide linkers and displayed on B. subtilis DB 403 spores. The successful display of CotG-nit fusion proteins on the spore surface of B. subtilis was verified by Western blot analysis and activity measurement. It was demonstrated that the fusion with linker GGGGSEAAAKGGGGS presented the highest thermal and pH stability, which is 2.67- and 1.9-fold of the fusion without linker...
2017: Journal of Molecular Microbiology and Biotechnology
Laura E Navas, Ariel F Amadio, Elio M Ortiz, Diego H Sauka, Graciela B Benintende, Marcelo F Berretta, Rubén O Zandomeni
We report the complete sequence and analysis of pFR260, a novel megaplasmid of 260,595 bp from the Bacillus thuringiensis strain INTA Fr7-4 isolated in Argentina. It carries 7 insecticidal genes: 3 cry8 copies previously reported, 2 vip1, and 2 vip2. Also, it carries a gene encoding a putative atypical Cry protein. These genes are arranged in a region of approximately 105 kbp in size with characteristics of a pathogenicity island with a potential coleopteran-specific insecticide profile. DNA strand composition asymmetry, as determined by GC skew analysis, and the presence of a Rep protein involved in the initiation of replication suggest a bidirectional theta mechanism of replication...
2017: Journal of Molecular Microbiology and Biotechnology
Xiaopeng Xu, Zuoming Nie, Zhiyong Zheng, Li Zhu, Xiaobei Zhan
This study aimed to investigate the effect of nitrogen sources on the production and rheological properties of welan gum produced by Sphingomonas sp. ATCC 31555. Six different nitrogen sources were used for ATCC 31555 fermentation, and 2 of these were further analyzed due to their more positive influence on welan gum production and bacterial biomass. Bacterial biomass, welan gum yield, welan viscosity, molecular weight, monosaccharide composition, acyl content, and welan structure were analyzed. Welan gum production and the biomass concentration of ATCC 31555 were higher in media containing NaNO3 and beef extract...
2017: Journal of Molecular Microbiology and Biotechnology
Darshna G Hirpara, Harsukh P Gajera, Hitesh Z Hirpara, Balubhai A Golakiya
The fungus Trichoderma is a teleomorph of the Hypocrea genus and associated with biological control of plant diseases. The microscopic, biochemical, and molecular characterization of Trichoderma was carried out and evaluated for in vitro antagonistic activity against the fungal pathogen Sclerotium rolfsii causing stem rot disease in groundnut. In total, 11 isolates of Trichoderma were examined for antagonism at 6 and 12 days after inoculation (DAI). Out of 11, T. virens NBAII Tvs12 evidenced the highest (87...
2017: Journal of Molecular Microbiology and Biotechnology
Sofía García, Noemí Flores, Ramón De Anda, Georgina Hernández, Guillermo Gosset, Francisco Bolívar, Adelfo Escalante
The culture of engineered Escherichia coli for shikimic acid (SA) production results in the synthesis of quinic acid (QA) and dehydroshikimic acid (DHS), reducing SA yield and impairing downstream processes. The synthesis of QA by quinate/shikimate dehydrogenase (YdiB, ydiB) has been previously proposed; however, the precise role for this enzyme in the production of QA in engineered strains of E. coli for SA production remains unclear. We report the effect of the inactivation or the overexpression of ydiB in E...
2017: Journal of Molecular Microbiology and Biotechnology
José-Luis Rodríguez-Mejía, Abigail Roldán-Salgado, Joel Osuna, Enrique Merino, Paul Gaytán
Recombinant protein expression is one of the key issues in protein engineering and biotechnology. Among the different models for assessing protein production and structure-function studies, green fluorescent protein (GFP) is one of the preferred models because of its importance as a reporter in cellular and molecular studies. In this research we analyze the effect of codon deletions near the amino terminus of different GFP proteins on fluorescence. Our study includes Gly4 deletions in the enhanced GFP (EGFP), the red-shifted GFP and the red-shifted EGFP...
2017: Journal of Molecular Microbiology and Biotechnology
Xilong Kang, Yun Yang, Yang Jiao, Hongqin Song, Li Song, Dan Xiong, Lili Wu, Zhiming Pan, Xinan Jiao
In 2009, a novel pandemic swine-origin influenza A (H1N1) virus caused a public emergency of international concern. Vaccination is the primary strategy for the control of influenza epidemics. However, the poor immunopotency of many vaccine antigens is a major barrier to the development of effective vaccines against influenza. Flagellin, a Toll-like receptor 5 (TLR5) ligand, has been used as an adjuvant to enhance the immunopotency of vaccines in preclinical studies. Here, we developed a recombinant candidate vaccine, HA1-2-fljB, in which the globular head of the hemagglutinin (HA) antigen (residues 62-284) from H1N1 virus was fused genetically to the N-terminus of Salmonella typhimurium fljB...
October 8, 2016: Journal of Molecular Microbiology and Biotechnology
Haitham Mohamed Amer
Human metapneumovirus (HMPV) is an important cause of respiratory tract illness in children. Two HMPV subgroups, A and B, and four genotypes, A1, A2, B1 and B2, have been identified. Concurrent circulation of the different genotypes in yearly epidemics has been recorded globally, but not in Saudi Arabia. The current report was designed to study HMPV epidemiology in Saudi children and to analyze the genetic diversity and circulation patterns. Nasopharyngeal aspirates (n = 174) were collected from hospitalized children in Riyadh (2008-2009)...
October 1, 2016: Journal of Molecular Microbiology and Biotechnology
Junrui Wang, Junli Zhang, Quan Fu, Sufang Guo, La Ta, Peng Sun
This study aimed to investigate the molecular mechanisms underlying the antibiotic resistance difference among three Acinetobacter baumannii isolates. Fifty A. baumannii isolates were first subjected to an antimicrobial susceptibility test, then three isolates differing in antibiotic resistance were selected and subjected to iTRAQ (isobaric tags for relative and absolute quantification)-based proteomics analysis. Differential proteins among the three A. baumannii isolates were further identified and subjected to gene ontology functional enrichment analysis...
September 16, 2016: Journal of Molecular Microbiology and Biotechnology
Masoumeh Douraghi, Seyedesomaye Jasemi, Mansoor Kodori, Mohammad Rahbar, Mohammad Ali Boroumand
Recent studies have recognized the ATPase-encoding comM gene as a hot spot for the integration of Acinetobacter baumannii resistance islands (RIs). Despite the circulation of high numbers of multidrug-resistant A. baumannii (MDR-AB) isolates in Middle East countries, no information is available about the interruption of comM and subsequent transposition into comM in isolates belonging to the global clones (GC) GC1, GC2, or GC3. In this study 401 A. baumannii isolates from hospitals in Tehran, Iran, were included...
September 16, 2016: Journal of Molecular Microbiology and Biotechnology
Naveen Kumar, Sanjay Barua, Riyesh Thachamvally, Bhupendra Nath Tripathi
Systems biology refers to system-wide changes in biological components such as RNA/DNA (genomics), protein (proteomics) and lipids (lipidomics). In this review, we provide comprehensive information about morbillivirus replication. Besides discussing the role of individual viral/host proteins in virus replication, we also discuss how systems-level analyses could improve our understanding of morbillivirus replication, host-pathogen interaction, immune response and disease resistance. Finally, we discuss how viroinformatics is likely to provide important insights for understanding genome-genome, genome-protein and protein-protein interactions...
September 9, 2016: Journal of Molecular Microbiology and Biotechnology
Takfarinas Kentache, Eliane Milohanic, Thanh Nguyen Cao, Abdelhamid Mokhtari, Francine Moussan Aké, Que Mai Ma Pham, Philippe Joyet, Josef Deutscher
Transposon insertion into Listeria monocytogenes lmo2665, which encodes an EIIC of the phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS), was found to prevent D-arabitol utilization. We confirm this result with a deletion mutant and show that Lmo2665 is also required for D-xylitol utilization. We therefore called this protein EIICAxl. Both pentitols are probably catabolized via the pentose phosphate pathway (PPP) because lmo2665 belongs to an operon, which encodes the three PTSAxl components, two sugar-P dehydrogenases, and most PPP enzymes...
August 24, 2016: Journal of Molecular Microbiology and Biotechnology
Thu Trang Thi Vu, Boram Jeong, Martin Krupa, Uijung Kwon, Jung-A Song, Bich Hang Do, Minh Tan Nguyen, Taewook Seo, Anh Ngoc Nguyen, Chul Hyun Joo, Han Choe
Human interferon alpha-2b (IFNα-2b) has therapeutic applications as an antiviral and antiproliferative drug and has been used for a wide range of indications. Efficient production of IFNα-2b in Escherichia coli has been difficult because the protein tends to form inclusion bodies. This obstacle has garnered interest in efficiently expressing IFNα-2b and overcoming its poor solubility. In this study, seven N-terminal fusion partners - hexahistidine (His6), thioredoxin, glutathione S-transferase (GST), maltose-binding protein (MBP), N-utilization substance protein A, protein disulfide bond isomerase (PDI), and b'a' domain of PDI - were tested for soluble overexpression of codon-optimized IFNα-2b in E...
July 28, 2016: Journal of Molecular Microbiology and Biotechnology
Harikrishnan Kuppusamykrishnan, Larry M Chau, Gabriel Moreno-Hagelsieb, Milton H Saier
We have designed a freely accessible program, PhyST, which allows the automated characterization of any family of homologous proteins within the Transporter Classification Database. The program performs an NCBI-PSI-BLAST search and reports (1) the average protein sequence length with standard deviations, (2) the average predicted number of transmembrane segments, (3) the total number of homologues retrieved, (4) a quantitative list of all source phyla, and (5) potential fusion proteins of sizes considerably exceeding the average size of the proteins retrieved...
2016: Journal of Molecular Microbiology and Biotechnology
Sarah Van Lent, Heather Huot Creasy, Garry S A Myers, Daisy Vanrompay
Variation is a central trait of the polymorphic membrane protein (Pmp) family. The number of pmp coding sequences differs between Chlamydia species, but it is unknown whether the number of pmp coding sequences is constant within a Chlamydia species. The level of conservation of the Pmp proteins has previously only been determined for Chlamydia trachomatis. As different Pmp proteins might be indispensible for the pathogenesis of different Chlamydia species, this study investigated the conservation of Pmp proteins both within and across C...
2016: Journal of Molecular Microbiology and Biotechnology
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