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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...
November 18, 2016: 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...
November 8, 2016: 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
Hanne L P Tytgat, Geert Schoofs, Jos Vanderleyden, Els J M Van Damme, Ruddy Wattiez, Sarah Lebeer, Baptiste Leroy
Glycoproteins form an interesting class of macromolecules involved in bacterial-host interactions, but they are not yet widely explored in Gram-positive and beneficial species. Here, an integrated and widely applicable approach was followed to identify putative bacterial glycoproteins, combining proteome fractionation with 2D protein and glycostained gels and lectin blots. This approach was validated for the microbiota isolate Lactobacillus rhamnosus GG. The approach resulted in a list of putative glycosylated proteins receiving a 'glycosylation score'...
2016: Journal of Molecular Microbiology and Biotechnology
Meriem Derkaoui, Ana Antunes, Jamila Nait Abdallah, Sandrine Poncet, Alain Mazé, Que Mai Ma Pham, Abdelhamid Mokhtari, Ala-Eddine Deghmane, Philippe Joyet, Muhamed-Kheir Taha, Josef Deutscher
We identified the genes encoding the proteins for the transport of glucose and maltose in Neisseria meningitidis strain 2C4-3. A mutant deleted for NMV_1892(glcP) no longer grew on glucose and deletion of NMV_0424(malY) prevented the utilization of maltose. We also purified and characterized glucokinase and α-phosphoglucomutase, which catalyze early catabolic steps of the two carbohydrates. N. meningitidis catabolizes the two carbohydrates either via the Entner-Doudoroff (ED) pathway or the pentose phosphate pathway, thereby forming glyceraldehyde-3-P and either pyruvate or fructose-6-P, respectively...
2016: Journal of Molecular Microbiology and Biotechnology
Neha Sharma, Nanda G Aduri, Anna Iqbal, Bala K Prabhala, Osman Mirza
Peptide transport in living organisms is facilitated by either primary transport, hydrolysis of ATP, or secondary transport, cotransport of protons. In this study, we focused on investigating the ligand specificity of the Neisseria meningitidis proton-coupled oligopeptide transporter (NmPOT). It has been shown that the gene encoding this transporter is upregulated during infection. NmPOT conformed to the typical chain length preference as observed in prototypical transporters of this family. In contrast to prototypical transporters, it was unable to accommodate a positively charged peptide residue at the C-terminus position of the substrate peptide...
2016: Journal of Molecular Microbiology and Biotechnology
Xiaofeng Ban, Caiming Li, Zhengbiao Gu, Chunhui Bao, Yijing Qiu, Yan Hong, Li Cheng, Zhaofeng Li
The branching enzyme (EC catalyzes the formation of α-1,6 branch points in starch. In this study, the Geobacillus thermoglucosidans gene-encoding branching enzyme was expressed in Escherichia coli BL21 (DE3) and the protein was isolated and characterized. G. thermoglucosidans branching enzyme is a thermostable enzyme with an optimal reaction temperature of nearly 60°C and a half-life at 65°C of approximately 1.1 h. The activity of the recombinant enzyme is optimal at pH 7.5, with broad stability between pH 5...
2016: Journal of Molecular Microbiology and Biotechnology
Hao Tan, Xiang Wu, Liyuan Xie, Zhongqian Huang, Weihong Peng, Bingcheng Gan
Four novel phytases of the histidine acid phosphatase family were identified in two publicly available metagenomic datasets of an acidic peat-soil microbiome in northeastern Bavaria, Germany. These enzymes have low similarity to all the reported phytases. They were overexpressed in Escherichia coli and purified. Catalytic efficacy in simulated gastric fluid was measured and compared among the four candidates. The phytase named rPhyPt4 was selected for its high activity. It is the first phytase identified from unculturable Acidobacteria...
2016: Journal of Molecular Microbiology and Biotechnology
Lin Liu, Xiangyan Zhang, Siyuan Yang, Yao Zhai, Weijia Liu, Xiaorong Wang, Zhao Zhang, Zhancheng Gao
AIMS: The aim of this project was to explore the different CTX-M expression levels occurring from a single conserved promoter with different spacer sequences, the variation of which is hypothesized to be a key factor in fluctuating levels of CTX-M. METHODS: The blaCTX-M promoter fragments with five different spacer sequences were amplified, sequenced and cloned into the pUA66 expression vector carrying the green fluorescent protein (GFP) gene. The expression of blaCTX-M in the transconjugants was analyzed using fluorescence microscopy, flow cytometry and qRT-PCR...
2016: Journal of Molecular Microbiology and Biotechnology
Fabien Baldacci-Cresp, Anaxi Houbaert, Amana Metuor Dabire, Adeline Mol, Daniel Monteyne, Mondher El Jaziri, Laurence Van Melderen, Marie Baucher
BACKGROUND/AIMS: The Escherichia coli MazF is an endoribonuclease that cleaves mRNA at ACA sequences, thereby triggering inhibition of protein synthesis. The aim of this study is to evaluate the efficiency of the mazEF toxin-antitoxin system in plants to develop biotechnological tools for targeted cell ablation. METHODS: A double transformation strategy, combining expression of the mazE antitoxin gene under the control of the CaMV 35S promoter, reported to drive expression in all plant cells except within the tapetum, together with the expression of the mazF gene under the control of the TA29 tapetum-specific promoter in transgenic tobacco, was applied...
2016: Journal of Molecular Microbiology and Biotechnology
Chandra Shekar Mootapally, Neelam M Nathani, Amrutlal K Patel, Subhash J Jakhesara, Chaitanya G Joshi
Phytases have been widely used as animal feed supplements to increase the availability of digestible phosphorus, especially in monogastric animals fed cereal grains. The present study describes the identification of a full-length phytase gene of Prevotella species present in Mehsani buffalo rumen. The gene, designated as RPHY1, consists of 1,251 bp and is expressed into protein with 417 amino acids. A homology search of the deduced amino acid sequence of the RPHY1 phytase gene in a nonredundant protein database showed that it shares 92% similarity with the histidine acid phosphatase domain...
2016: Journal of Molecular Microbiology and Biotechnology
Jinfeng Shao, Marcelo F M Marcondes, Vitor Oliveira, Jaap Broos
Chemically defined media for growth of Lactococcus lactis strains contain about 50 components, making them laborious and expensive growth media. However, they are crucial for metabolism studies as well as for expression of heterologous proteins labeled with unnatural amino acids. In particular, the L. lactis Trp auxotroph PA1002, overexpressing the tryptophanyl tRNA synthetase enzyme of L. lactis, is very suitable for the biosynthetic incorporation of Trp analogs in proteins because of its most relaxed substrate specificity reported towards Trp analogs...
2016: Journal of Molecular Microbiology and Biotechnology
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