Phage display-derived inhibitor of the essential cell wall biosynthesis enzyme MurF

Background To develop antibacterial agents having novel modes of action against bacterial cell wall biosynthesis, we targeted the essential MurF enzyme of the antibiotic resistant pathogen Pseudomonas aeruginosa. MurF catalyzes the formation of a peptide bond between D-Alanyl-D-Alanine (D-Ala-D-Ala) and the cell wall precursor uridine 5'-diphosphoryl N-acetylmuramoyl-L-alanyl-D-glutamyl-meso-diaminopimelic acid (UDP-MurNAc-Ala-Glu-meso-A2pm) with the concomitant hydrolysis of ATP to ADP and inorganic phosphate, yielding UDP-N-acetylmuramyl-pentapeptide. As MurF acts on a dipeptide, we exploited a phage display approach to identify peptide ligands having high binding affinities for the enzyme. Results Screening of a phage display 12-mer library using purified P. aeruginosa MurF yielded to the identification of the MurFp1 peptide. The MurF substrate UDP-MurNAc-Ala-Glumeso-A2pm was synthesized and used to develop a sensitive spectrophotometric assay to quantify MurF kinetics and inhibition. MurFp1 acted as a weak, time-dependent inhibitor of MurF activity but was a potent inhibitor when MurF was pre-incubated with UDP-MurNAc-Ala-Glu-meso-A2pm or ATP. In contrast, adding the substrate D-Ala-D-Ala during the pre-incubation nullified the inhibition. The IC50 value of MurFp1 was evaluated at 250 μM, and the Ki was established at 420 μM with respect to the mixed type of inhibition against D-Ala-D-Ala. Conclusion MurFp1 exerts its inhibitory action by interfering with the utilization of D-Ala-D-Ala by the MurF amide ligase enzyme. We propose that MurFp1 exploits UDP-MurNAc-Ala-Glu-meso-A2pm-induced structural changes for better interaction with the enzyme. We present the first peptide inhibitor of MurF, an enzyme that should be exploited as a target for antimicrobial drug development

Le Forum, Vol. 40 No. 2

https://digitalcommons.library.umaine.edu/francoamericain_forum/1087/thumbnail.jp

Le FORUM, Vol. 36 No. 4

https://digitalcommons.library.umaine.edu/francoamericain_forum/1035/thumbnail.jp

Le FORUM, Vol. 38 No. 2

https://digitalcommons.library.umaine.edu/francoamericain_forum/1041/thumbnail.jp

Le Forum, Vol. 40 No. 3

https://digitalcommons.library.umaine.edu/francoamericain_forum/1088/thumbnail.jp

Using Acoustic Sensors to Improve the Efficiency of the Forest Value Chain in Canada: A Case Study with Laminated Veneer Lumber

Engineered wood products for structural use must meet minimum strength and stiffness criteria. This represents a major challenge for the industry as the mechanical properties of the wood resource are inherently variable. We report on a case study that was conducted in a laminated veneer lumber (LVL) mill in order to test the potential of an acoustic sensor to predict structural properties of the wood resource prior to processing. A population of 266 recently harvested aspen logs were segregated into three sub-populations based on measurements of longitudinal acoustic speed in wood using a hand tool equipped with a resonance-based acoustic sensor. Each of the three sub-populations were peeled into veneer sheets and graded for stiffness with an ultrasonic device. The average ultrasonic propagation time (UPT) of each subpopulation was 418, 440 and 453 microseconds for the green, blue, and red populations, respectively. This resulted in contrasting proportions of structural veneer grades, indicating that the efficiency of the forest value chain could be improved using acoustic sensors. A linear regression analysis also showed that the dynamic modulus of elasticity (MOE) of LVL was strongly related to static MOE (R2 = 0.83), which suggests that acoustic tools may be used for quality control during the production process

Persistent anthrax as a major driver of wildlife mortality in a tropical rainforest

Anthrax is a globally important animal disease and zoonosis. Despite this, our current knowledge of anthrax ecology is largely limited to arid ecosystems, where outbreaks are most commonly reported. Here we show that the dynamics of an anthrax-causing agent, Bacillus cereus biovar anthracis, in a tropical rainforest have severe consequences for local wildlife communities. Using data and samples collected over three decades, we show that rainforest anthrax is a persistent and widespread cause of death for a broad range of mammalian hosts. We predict that this pathogen will accelerate the decline and possibly result in the extirpation of local chimpanzee (Pan troglodytes verus) populations. We present the epidemiology of a cryptic pathogen and show that its presence has important implications for conservation

Le FORUM, Vol. 38 No. 3

https://digitalcommons.library.umaine.edu/francoamericain_forum/1042/thumbnail.jp

Molecular phylogenetics and temporal diversification in the genus Aeromonas based on the sequences of five housekeeping genes

Several approaches have been developed to estimate both the relative and absolute rates of speciation and extinction within clades based on molecular phylogenetic reconstructions of evolutionary relationships, according to an underlying model of diversification. However, the macroevolutionary models established for eukaryotes have scarcely been used with prokaryotes. We have investigated the rate and pattern of cladogenesis in the genus Aeromonas (γ-Proteobacteria, Proteobacteria, Bacteria) using the sequences of five housekeeping genes and an uncorrelated relaxed-clock approach. To our knowledge, until now this analysis has never been applied to all the species described in a bacterial genus and thus opens up the possibility of establishing models of speciation from sequence data commonly used in phylogenetic studies of prokaryotes. Our results suggest that the genus Aeromonas began to diverge between 248 and 266 million years ago, exhibiting a constant divergence rate through the Phanerozoic, which could be described as a pure birth process