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Identification of MazF Homologue in Legionella pneumophila Which Cleaves RNA at the AACU Sequence
MazF is a sequence-specific endoribonuclease that is widely conserved in bacteria and archaea. Here, we found an MazF homologue (MazF-lp; LPO-p0114) inLegionella pneumophila. The mazF-lp gene overlaps 14 base pairs with the upstream gene mazE-lp (MazE-lp; LPO-p0115). The induction ofmazF-lp caused cell growth arrest, whilemazE-lp co-induction recovered cell growth inEscherichia coli. In vivo and in vitro primer extension experiments showed that MazF-lp is a sequence-specific endoribonuclease cleaving RNA at AACU. The endoribonuclease activity of purified MazF-lp was inhibited by purified MazE-lp. We found that MazE-lp and ...
Source: Journal of Molecular Microbiology and Biotechnology - March 20, 2019 Category: Microbiology Source Type: research

Cytochrome bcc-aa3 Oxidase Supercomplexes in the Aerobic Respiratory Chain of Streptomyces coelicolor A3(2)
Streptomyces coelicolor A3(2), an obligately aerobic, oxidase-positive, and filamentous soil bacterium, lacks a soluble cytochromec in its respiratory chain, having instead a membrane-associated dihemec-type cytochrome, QcrC. This necessitates complex formation to allow electron transfer between the cytochromebcc andaa3 oxidase respiratory complexes. Combining genetic complementation studies with in-gel cytochrome oxidase activity staining, we demonstrate that the completeqcrCAB-ctaCDFE gene locus on the chromosome, encoding, respectively, thebcc andaa3 complexes, is required to manifest a cytochrome oxidase enzyme activit...
Source: Journal of Molecular Microbiology and Biotechnology - March 15, 2019 Category: Microbiology Source Type: research

Fast Biodegradation of Diesel Hydrocarbons at High Concentration by the Sophorolipid-Producing Yeast Candida catenulata KP324968
In the last decades, biodegradation as an environmentally friendly approach has raised interest in connection with the removal of hydrocarbon pollutants. Its capacity for removing pollutants strongly depends on the type of living cell and environmental conditions. The degradative activity of a new sophorolipid-producing yeast,Candida catenulata KP324968, in the removal of high concentrations of diesel from effluents was statistically evaluated considering the initial pH, the agitation speed, and the initial diesel concentration. The optimal setting of the operational variables at an initial pH of 4.7, an agitation speed of...
Source: Journal of Molecular Microbiology and Biotechnology - March 8, 2019 Category: Microbiology Source Type: research

Relationship between Adaptive Changing of Lysophosphatidylethanolamine Content in the Bacterial Envelope and Ampicillin Sensitivity of Yersinia pseudotuberculosis 
The low permeability of porin channels is the possible reason for Gram-negative bacterial resistance to antibiotics. The adaptive accumulation of lysophosphatidylethanolamine (LPE) inYersinia pseudotuberculosis induces conformational changes of OmpF porin that may hinder the transport of antibiotics through this channel. The present study was aimed to test whether the changes in LPE content affect the resistance of bacteria to ampicillin. The addition of glucose to the culture medium was shown to simultaneously increase the level of LPE and minimum inhibitory concentration (MIC) for ampicillin ofY. pseudotuberculosis cells...
Source: Journal of Molecular Microbiology and Biotechnology - March 8, 2019 Category: Microbiology Source Type: research

Mutation in afsR Leads to A-Factor Deficiency in Streptomyces griseus B2682
Conclusions: AfsR has an effect on the regulation of A-factor production inS. griseus. The presence of AfsA is not sufficient for normal A-factor production. AfsR regulates A-factor biosynthesis independently of AfsA.J Mol Microbiol Biotechnol 2018;28:216 –224 (Source: Journal of Molecular Microbiology and Biotechnology)
Source: Journal of Molecular Microbiology and Biotechnology - February 19, 2019 Category: Microbiology Source Type: research

Production of Polyhydroxyalkanoates Copolymers by Recombinant Pseudomonas in Plasmid- and Antibiotic-Free Cultures
We reported for the first time the insertion of a chromosomal copy ofphaC using the transposon system mini-Tn7. Stable antibiotic marker-free and plasmid-free recombinants were obtained. Subsequently, P(3HB-co-3HAMCL) was produced by these recombinants using glucose as the sole carbon source, without the need for co-substrates and under antibiotic-free conditions. A recombinant harboringA. hydrophila phaC produced a terpolyester composed of 84.2 mol% of 3-hydroxybutyrate, 6.3 mol% of 3-hydroxyhexanoate, and 9.5 mol% of 3-hydroxydecanoate from only glucose. Hence, we were successful in increasing the industrial potential of...
Source: Journal of Molecular Microbiology and Biotechnology - February 19, 2019 Category: Microbiology Source Type: research

Molecules Autoinducer 2 and cjA and Their Impact on Gene Expression in Campylobacter jejuni 
Quorum sensing is a widespread form of cell-to-cell communication, which is based on the production of signaling molecules known as autoinducers (AIs). The first group contains highly species-specific N-acyl homoserine lactones (N-AHLs), generally known as AI-1, which are produced by AHL synthase. The second group, possessing the characteristic structure of a furanone ring, are known as AI-2. The enzyme responsible for their production is S-ribosylhomocysteine lyase (LuxS). InCampylobacter jejuni, AI-2 and LuxS play a role in many important processes, including biofilm formation, stress response, motility, expression of vi...
Source: Journal of Molecular Microbiology and Biotechnology - January 29, 2019 Category: Microbiology Source Type: research

Multidrug Resistance and the Predominance of bla CTX-M in Extended Spectrum Beta-Lactamase-Producing Enterobacteriaceae of Animal and Water Origin
The aim of this work was the genetic characterization of cefotaxime-resistant enterobacteria from animals (53 samples), the surface water of rivers (17 samples), and wastewater treatment plants (43 samples) in Tunisia. A total of 48 (42.4%) cefotaxime-resistant isolates were recovered. An extended spectrum beta-lactamase (ESBL) phenotype with a positive double-disk synergy test (DDST) was exhibited by 34 (70.8%) and 14 (29.1%) isolates from water and animal origins, respectively. Isolates from water were identified as:Escherichia coli (n = 17),Hafnia spp. (n = 13),Citrobacter spp. (n = 1),Enterobacter cloacae (n = 1),Klebs...
Source: Journal of Molecular Microbiology and Biotechnology - January 4, 2019 Category: Microbiology Source Type: research

Overexpression of an Inulinase Gene in an Oleaginous Yeast, Aureobasidium melanogenum P10, for Efficient Lipid Production from Inulin
In this study, in order to directly and efficiently convert inulin into a single-cell oil (SCO), anINU1 gene encoding inulinase fromKluyveromyces marxianus was integrated into the genomic DNA and actively expressed in an SCO producerAureobasidiummelanogenum P10. The transformant API41 obtained produced 28.5 U/mL of inulinase and its wild-type strain P10 yielded only 8.62 U/mL. Most (97.5%) of the inulinase produced by the transformant API41 was secreted into the culture. During a 10-L fermentation, 66.2% (w/w) lipid in the yeast cells of the transformant API41 and 14.38 g/L of cell dry weight were attained from inulin of 8...
Source: Journal of Molecular Microbiology and Biotechnology - January 3, 2019 Category: Microbiology Source Type: research

Expression of Cell-Penetrating Peptides Fused to Protein Cargo
Cell-penetrating peptides (CPPs) are short peptides that can cross cell membranes. CPPs enable the delivery of biomolecules into cells and can act as drug-delivery vectors. Because recombinant production of CPPs as fusions to protein “cargo” leads to low yields for some CPP-cargo fusions, approaches to enhance the recombinant expression of peptide-cargo fusions need to be identified. We optimized expression conditions inEscherichia coli for fusions of CPPs (SynB, histatin-5, and MPG) to the cargo proteins biotin carboxyl carrier protein, maltose-binding protein, and green fluorescent protein. We used Western blotting t...
Source: Journal of Molecular Microbiology and Biotechnology - December 19, 2018 Category: Microbiology Source Type: research

A Simple Method for Removal of the Chlamydomonas reinhardtii Cell Wall Using a Commercially Available Subtilisin (Alcalase)
The algal cell wall is a potent barrier for delivery of transgenes for genetic engineering. Conventional methods developed for higher plant systems are often unable to penetrate or remove algal cell walls owing to their unique physical and chemical properties. Therefore, we developed a simple transformation method forChlamydomonas reinhardtii using commercially available enzymes. Out of 7 enzymes screened for cell wall disruption, a commercial form of subtilisin (Alcalase) was the most effective at a low concentration (0.3 Anson units/mL). The efficiency was comparable to that of gamete lytic enzyme, a protease commonly us...
Source: Journal of Molecular Microbiology and Biotechnology - December 19, 2018 Category: Microbiology Source Type: research

Subunit Composition of Ribosome in the yqgF Mutant Is Deficient in pre-16S rRNA Processing of Escherichia coli 
Escherichia coli 16S, 23S, and 5S ribosomal RNAs (rRNAs) are transcribed as a single primary transcript, which is subsequently processed into mature rRNAs by several RNases. Three RNases (RNase III, RNase E, and RNase G) were reported to function in processing the 5 ′-leader of precursor 16S rRNA (pre-16S rRNA). Previously, we showed that a novel essential YqgF is involved in that processing. Here we investigated the ribosome subunits of theyqgFts mutant by LC-MS/MS. The mutant ribosome had decreased copy numbers of ribosome protein S1, suggesting that theyqgF gene enables incorporation of ribosomal protein S1 into ribos...
Source: Journal of Molecular Microbiology and Biotechnology - December 19, 2018 Category: Microbiology Source Type: research

Identification of A Ring-Hydroxylating Dioxygenases Capable of Anthracene and Benz[a]anthracene Oxidization from Rhodococcus sp. P14
Nowadays, contamination of soil and marine sediments by polycyclic aromatic hydrocarbons (PAHs) has become a serious problem all over the world.Rhodococcus sp. P14 was isolated from sediments with crude oil contaminate and showed degradation ability on various PAHs. The genome ofRhodococcus sp. P14 was sequenced. A gene cluster encoding a ring-hydroxylating dioxygenase Baa related to PAH degradation was identified by bioinformatics. The expression level of genebaaA was increased when P14 was cultured with anthracene, pyrene, phenanthrene, or benz[a] anthracene as the single carbon source. The recombinant protein Baa was ...
Source: Journal of Molecular Microbiology and Biotechnology - December 19, 2018 Category: Microbiology Source Type: research

Development and Application of a New PCR Method for Detection of Blumeria graminis f. sp. tritici 
We developed new PCR assays that target beta-tubulin (TUB2) and 14 alpha-demethylase (CYP51) genes and used them for the species-specific detection ofBlumeria graminis f. sp.tritici (Bgt). Based on fungi DNA sequences available in the NCBI (National Center for Biotechnology Information) GenBank database we developed simplex and duplex PCR assays. The specificities of the primer sets were evaluated using environmental samples of wheat leaves collected during the 2015/2016 growing season across Poland. Primer sets LidBg17/18 and LidBg21/22 strongly amplified fragments of the expected length for all 67 tested samples. Primer ...
Source: Journal of Molecular Microbiology and Biotechnology - December 6, 2018 Category: Microbiology Source Type: research

Photoinactivation of the Staphylococcus aureus Lactose-Specific EIICB Phosphotransferase Component with p -azidophenyl- β-D-Galactoside and Phosphorylation of the Covalently Bound Substrate
Conclusion: Phosphorylation of the –OH group at C6 ofp-nitrenephenyl- β-D-galactopyranoside covalently bound to EIICLac by the histidyl-phosphorylated [32P]P ∼EIIBLac domain is a likely explanation for the observed acid resistance. Placingp-nitrenephenyl- β-D-galactopyranoside into the active site of modelled EIICLac suggested that the nitrene binds to the -NH- group of Ser248, which would explain why no sequence data beyond Pro247could be obtained.J Mol Microbiol Biotechnol 2018;28:147 –158 (Source: Journal of Molecular Microbiology and Biotechnology)
Source: Journal of Molecular Microbiology and Biotechnology - December 6, 2018 Category: Microbiology Source Type: research