Fems Microbiology Letters最新文献

Pseudomonas aeruginosa is a remarkably adaptive bacterium frequently implicated in severe, sometimes fatal infections within healthcare institutions. The origins of clinical strains have generated considerable debate, suggesting that infectious variants emerge through selection from a broader environmental bacterial population. Our investigation explored the physiological differences between environmental (indoor) and clinical strains of P. aeruginosa isolated from a hospital over a year. We assessed various parameters, including survival patterns, antibiotic resistance, vulnerability to ciliate predation, bacterial antagonism, and motility. Despite the minimal incidence of patient infections during our observation, environmental P. aeruginosa was prevalent throughout the hospital during our study. Clinical strains exhibited diminished resistance to certain antibiotics, increased resistance against ciliate predators, and enhanced swarming and swimming motility compared to their environmental counterparts. Clinical strains maintained higher cell densities under starvation conditions but were outcompeted by environmental strains in a nutrient medium. In conclusion, our study suggests that P. aeruginosa clinical isolates possess unique physiological adaptations that may favor host colonization.

Although 30 years have passed since the description of sugar-induced cell death (SICD), the specific molecular mechanism that triggers this process remains unclear. This paper attempts to shed light on the relationship between SICD and glucose catabolism. In yeast cells, glucose is involved not only in energy-producing processes but also in the synthesis of reserve hydrocarbons. It is known that disruption of trehalose synthesis leads to significant changes in the physiology of S. cerevisiae. The present study shows that deletion of the TPS1 gene resulted in a 44% suppression of SICD and a 75% reduction in the number of cells with excess ROS. The suppression was comparable to the suppression of SICD (38%) and ROS (71%) with deletion of the HXK2 gene. Since HXK2 is the first enzyme in the glycolytic pathway, the effect of two other key glycolytic enzymes on SICD was tested. Deletion of the TDH3 gene (glyceraldehyde-3-phosphate dehydrogenase) resulted in a 39% suppression of SICD and ROS by 48%. Inhibition of Tdh3p with 1 mM iodoacetamide also suppressed SICD by 67% and ROS by 58%. Deletion of the PFK1 (phosphofructokinase 1) gene resulted in a complete block of SICD (97%) but unexpectedly resulted in a significant increase in the number of cells with excess ROS. The results obtained suggest that such a phenomenon as SICD is the result of an imbalance in the cellular pathways of glucose catabolism.

The peptidoglycan stem peptides of the hyperthermophile Thermotoga maritima contain an unusual D-lysine (D-Lys) alongside the usual D-alanine and D-glutamate. We identified a Lys racemase that catalyzes racemization between L-Lys and D-Lys, and a diaminopimelate (Dpm) epimerase that catalyzes epimerization between LL-Dpm and meso-Dpm. Herein, we characterized a Dpm decarboxylase (TM1517) that catalyzes the conversion of meso-Dpm to L-Lys. TM1517 displayed high decarboxylase activity toward meso-Dpm but no activity toward LL-Dpm. D-Lys was not detected in the decarboxylation of meso-Dpm. The pH and temperature dependencies and kinetic parameters of decarboxylase activity were determined. Although other amino acid metabolizing activities of TM1517 were investigated, TM1517 did not exhibit any activities. Therefore, TM1517 is a Dpm decarboxylase associated with L- and D-Lys biosynthesis in T. maritima.

Carboxylic acid reductases (CARs) have been garnering attention in applications for the sustainable synthesis of aldehydes. Despite numerous discoveries, not all characteristics of CAR enzymes have been extensively studied or understood. Herein, we report the discovery and expression of a new CAR enzyme (TvirCAR2) from the ascomycetous fungus, Trichoderma virens. Tvircar2 is one of five putative CARs identified from analyses of the T. virens genome. In silico analyses showed that TvirCAR2 has a high hydrophobicity index and that its corresponding gene is part of a biosynthetic gene cluster predicted to synthesize hybrid PKS-NRPS secondary metabolites. TvirCAR2 was highly expressed as soluble and insoluble forms in an Escherichia coli expression host. The solubility of the purified TvirCAR2 necessitated the addition of glycerol in the purification and assay buffers. Substrate screening via molecular docking showed that benzoic acid was a suitable substrate candidate. The TvirCAR2 enzyme catalyzed the reduction of benzoic acid with a specific activity of around 1.4 µmol/h/mg. Homologs which are predicted to exhibit similar hydrophobicity are the CARs from Stachybotrys bisbyi (StbB) which is involved in the production of the meroterpenoid, ilicicolin B, and Trichoderma reesei (TrCAR) which is part of a similar but still uncharacterized biosynthetic gene cluster.

Scientific conferences are essential to academic exchange. However, related air travel contributes to greenhouse gas emissions, while expensive registration and travel costs limit the participation of early-career researchers and those from low-income countries. Virtual conferences offer promising solutions for reducing emissions and enhancing accessibility and inclusivity but often limit networking and personal interaction. Hybrid multi-hub conferences, which combine virtually connected in-person venues with individual virtual participation, combine the benefits of both conference formats. Thus, we present and discuss MEEhubs2024, a multi-hub conference on microbial ecology and evolution held in January 2024. During this three-day conference, attendees participated virtually or at one of six hubs in Europe and North America. We analyzed the participants' and organizers' feedback to create a template and provide insights into the scientific community's adoption of this new conference format, which was positively evaluated by most participants. Because technical, logistical and structural challenges remain, including limited opportunities to interact and network across hubs and participation modes, we provide recommendations for improvement like hiring technical hosts and offering virtual-only social activities. Finally, we used the participants' feedback to reflect on conference expectations, highlighting research gaps and the need for organizers to define and communicate goals when organizing conferences.

Antibiotic resistance and the persistence of sessile cells within biofilms complicate the eradication of biofilm-related infections using conventional antibiotics. This highlights the necessity for alternate therapy methods. The objective of this study was to investigate the biofilm destruction activity of α-tocopherol against Staphylococcus aureus, Proteus mirabilis and Pseudomonas aeruginosa on polystyrene. α-Tocopherol showed significant biofilm destruction activity on the pre-formed biofilms of S. aureus (45-46%), Pr. mirabilis (42-54%) and Ps. aeruginosa (28%). Resazurin assay showed that α-tocopherol disrupted all bacteria biofilms without interfering with their cell viability. Scanning electron microscope images showed lower bacterial cell count and less compacted cell aggregates on polystyrene surfaces after treatment with alpha-tocopherol. This study demonstrated the biofilm destruction activity of alpha-tocopherol against S. aureus, Pr. mirabilis and Ps. aeruginosa. α-Tocopherol could potentially be used to decrease biofilm-associated infections of these bacteria.

This study assesses halophilic archaea's phylogenetic diversity in southern Tunisia's geothermal water. In the arid southern regions, limited surface freshwater resources make geothermal waters a vital source for oases and greenhouse irrigation. Three samples, including water, sediment, and halite soil crust, were collected downstream of two geothermal springs of the Ksar Ghilane (KGH) and Zaouet Al Aness (ZAN) oases, Tunisia. The samples were subjected to 16S rRNA gene sequencing using the Illumina Miseq sequencing approach. Several haloarchaea were identified in the geothermal springs. The average taxonomic composition revealed that 20 out of 33 genera were shared between the two geothermal sources, with uneven distribution, where the Halogranum genus was the most represented genus with an abundance of 18.9% and 11.58% for ZAW and KGH, respectively. Several unique site-specific genera were observed: Halonotius, Halopelagius, Natronorubrum, and Haloarcula in ZAN, and Haloprofundus, Halomarina, Halovivax, Haloplanus, Natrinema, Halobium, Natronoarchaeum, and Haloterrigena in the KGH pool. Most genus members are typically found in low-salinity ecosystems. These findings suggest that haloarchaea can disperse downstream from geothermal sources and may survive temperature and chemical fluctuations in the runoff.

Aeration is a common pretreatment method to enhance biogas production via anaerobic digestion of waste organic feedstocks such as unused food. While impacts on downstream anaerobic digestion have been intensively investigated, the consequence of aeration on the microbial community in food waste has not been characterized. Food waste has a low pH resulting from the dominance of lactic acid bacteria within the Firmicutes phylum. This excludes other phylotypes with a higher potential to hydrolyse complex biopolymers in food waste. In this study, we reveal that aeration of macerated food waste results in a dramatic shift away from Firmicutes towards dominance of Proteobacteria that are better known for extracellular enzyme production. Given that hydrolysis is the rate limiting step in anaerobic digestion, this explains why aeration improves the efficiency of biogas production from food waste. The discovery that Proteobacteria dominate microbial communities in aerated food waste opens up opportunities to manipulate extracellular enzyme production through gene expression mechanisms common among Proteobacteria such as quorum sensing.

Commensal Neisseria are members of a healthy human oropharyngeal microbiome; however, they also serve as a reservoir of antimicrobial resistance for their pathogenic relatives. Despite their known importance as sources of novel genetic variation for pathogens, we still do not understand the full suite of resistance mutations commensal species can harbor. Here, we use in vitro selection to assess the mutations that emerge in response to ciprofloxacin selection in commensal Neisseria by passaging four replicates of four different species in the presence of a selective antibiotic gradient for 20 days; then categorized derived mutations with whole genome sequencing. Ten out of sixteen selected cells lines across the four species evolved ciprofloxacin resistance (≥1 ug/ml); with resistance-contributing mutations primarily emerging in DNA gyrase subunit A and B (gyrA and gyrB), topoisomerase IV subunits C and E (parC and parE), and the multiple transferable efflux pump repressor (mtrR). Of note, these derived mutations appeared in the same loci responsible for ciprofloxacin-reduced susceptibility in the pathogenic Neisseria, suggesting conserved mechanisms of resistance across the genus. Additionally, we tested for zoliflodacin cross-resistance in evolved strain lines and found 6 lineages with elevated zoliflodacin minimum inhibitory concentrations. Finally, to interrogate the likelihood of experimentally derived mutations emerging and contributing to resistance in natural Neisseria, we used a population-based approach and identified GyrA 91I as a substitution circulating within commensal Neisseria populations and ParC 85C in a single gonococcal isolate. A small cluster of gonococcal isolates shared commensal alleles at parE, suggesting recent cross-species recombination events.

Terminal olefins are important platform chemicals, drop-in compatible hydrocarbons and also play an important role as biocontrol agents of plant pathogens. Currently, 1-alkenes are derived from petroleum, although microbial biosynthetic routes are known. Jeotgalicoccus sp. ATCC 8456 produces 1-alkenes via the fatty acid decarboxylase OleTJE. UndA and UndB are recently identified non-heme iron oxidases converting medium-chain fatty acids into terminal alkenes. Our knowledge about the diversity and natural function of OleTJE, UndA, and UndB homologs is scarce. We applied a combined screening strategy-solid-phase microextraction coupled with gas chromatography-mass spectrometry (SPME GC-MS) and polymerase chain reaction (PCR)-based amplification-to survey an environmental strain collection for microbial 1-alkene producers and their corresponding enzymes. Our results reinforce the high level of conservation of UndA and UndB genes across the genus Pseudomonas. In vivo production of defined 1-alkenes (C9-C13; C15; C19) was directed by targeted feeding of fatty acids. Lauric acid feeding enabled 1-undecene production to a concentration of 3.05 mg l-1 in Jeotgalicoccus sp. ATCC 8456 and enhanced its production by 105% in Pseudomonas putida 1T1 (1.10 mg l-1). Besides, whole genome sequencing of Jeotgalicoccus sp. ATCC 8456 enabled reconstruction of the 1-alkene biosynthetic pathway. These results advance our understanding of microbial 1-alkene synthesis and the underlying genetic basis.