Bacterial multi-solute transporters

Bacterial membrane proteins of the SbmA/BacA family are multi-solute transporters that mediate the uptake of structurally diverse hydrophilic molecules, including aminoglycoside antibiotics and antimicrobial peptides. Some family members are full-length ATP-binding cassette (ABC) transporters, whereas other members are truncated homologues that lack the nucleotide-binding domains and thus mediate ATP-independent transport. A recent cryo-EM structure of the ABC transporter Rv1819c from Mycobacterium tuberculosis has shed light on the structural basis for multi-solute transport and has provided insight into the mechanism of transport. Here, we discuss how the protein architecture makes SbmA/BacA family transporters prone to inadvertent import of antibiotics and speculate on the question which physiological processes may benefit from multi-solute transport

Amylomaltase of Pyrobaculum aerophilum IM2 produces thermoreversible starch gels

Amylomaltases are 4-α-glucanotransferases (EC 2.4.1.25) of glycoside hydrolase family 77 that transfer α-1,4-linked glucans to another acceptor, which can be the 4-OH group of an α-1,4-linked glucan or glucose. The amylomaltase-encoding gene (PAE1209) from the hyperthermophilic archaeon Pyrobaculum aerophilum IM2 was cloned and expressed in Escherichia coli, and the gene product (PyAMase) was characterized. PyAMase displays optimal activity at pH 6.7 and 95°C and is the most thermostable amylomaltase described to date. The thermostability of PyAMase was reduced in the presence of 2 mM dithiothreitol, which agreed with the identification of two possible cysteine disulfide bridges in a three-dimensional model of PyAMase. The kinetics for the disproportionation of malto-oligosaccharides, inhibition by acarbose, and binding mode of the substrates in the active site were determined. Acting on gelatinized food-grade potato starch, PyAMase produced a thermoreversible starch product with gelatin-like properties. This thermoreversible gel has potential applications in the food industry. This is the first report on an archaeal amylomaltase

Prospective CO2 and CO bioconversion into ectoines using novel microbial platforms

Microbial conversion of CO2 and CO into chemicals is a promising route that can contribute to the cost-effective reduction of anthropogenic green house and waste gas emissions and create a more circular economy. However, the biotechnological valorization of CO2 and CO into chemicals is still restricted by the limited number of model microorganisms implemented, and the small profit margin of the products synthesized. This perspective paper intends to explore the genetic potential for the microbial conversion of CO2 and CO into ectoines, in a tentative to broaden bioconversion platforms and the portfolio of products from C-1 gas fermentations. Ectoine and hydroxyectoine can be produced by microorganisms growing at high salinity. They are high-value commodities for the pharmaceutical and medical sectors (1000-1200 euro/kg). Currently microbial ectoine production is based on sugar fermentations, but expansion to other more sustainable and cheaper substrates is desirable. In this work, a literature review to identify halophilic microbes able to use CO2 and CO as a carbon source was performed. Subsequently, genomes of this poll of microbes were mined for genes that encode for ectoine and hydroxyectoine synthesis (ectABCD, ask, asd and ask_ect). As a result, we identified a total of 31 species with the genetic potential to synthesize ectoine and 14 to synthesize hydroxyectoine. These microbes represent the basis for the creation of novel microbial-platforms that can promote the development of cost-effective and sustainable valorization chains of CO2 and CO in different industrial scenarios

Incorporating needs-satisfaction in a discrete choice model of leisure activities

In this paper we extend the behavioural scope of discrete choice models for leisure activity-travel choices. More specifically, we investigate to what extent choices for leisure activities and related travels are driven by the satisfaction of needs. In addition to conventional attributes (such as activity costs), our regret based discrete choice model incorporates latent variables representing the anticipated level of individual needs-satisfaction by a particular leisure activity. The latent variables are calibrated with the help of subjective indicators of needs-satisfaction associated with the leisure activities. Results show that needs-satisfaction allows us to decompose a substantial share of the unobserved heterogeneity in leisure activity-travel decisions across respondents. Identifying the structural drivers of anticipated needs-satisfaction also enables a better prediction of leisure activity choice

A closed Candidatus Odinarchaeum chromosome exposes Asgard archaeal viruses

Asgard archaea have recently been identified as the closest archaeal relatives of eukaryotes. Their ecology, and particularly their virome, remain enigmatic. We reassembled and closed the chromosome of Candidatus Odinarchaeum yellowstonii LCB_4, through long-range PCR, revealing CRISPR spacers targeting viral contigs. We found related viruses in the genomes of diverse prokaryotes from geothermal environments, including other Asgard archaea. These viruses open research avenues into the ecology and evolution of Asgard archaea

Educational intervention to increase nurses' knowledge, self-efficacy and usage of telehealth:A multi-setting pretest-posttest study

The widespread use of telehealth, providing healthcare remotely, is hampered by various barriers. Dutch nurses currently working in practice never received education in this new way of healthcare delivery. Education is frequently suggested as a strategy to overcome barriers in telehealth use. However, the nature and effectiveness of such education has not yet been specified and tested in practice. In a previous study, we identified 14 nursing telehealth activities and accompanying competencies. In the current study, we established the effectiveness of training in these competencies on nurses' subjective knowledge, self-efficacy and usage of telehealth. A two-day tailored training program in nursing telehealth activities was evaluated in a Dutch context among 37 participants across three settings: (a) twelve primary care (PC), (b) fourteen homecare (HC) and (c) eleven hospital (H) nurses. In each team, telehealth knowledge significantly increased during the training sessions. In each team, nurses' telehealth self-efficacy also significantly increased 6-10 weeks after the training. After the training, the number of remote consultations increased from 2 to 12 in primary care, 12 to 35 in homecare and decreased from 28 to 17 in the hospital setting. We conclude that training nurses in telehealth activities contributes to their knowledge and self-efficacy.</p

Chlamydial contribution to anaerobic metabolism during eukaryotic evolution

The origin of eukaryotes is a major open question in evolutionary biology. Multiple hypotheses posit that eukaryotes likely evolved from a syntrophic relationship between an archaeon and an alphaproteobacterium based on H-2 exchange. However, there are no strong indications that modern eukaryotic H-2 metabolism originated from archaea or alphaproteobacteria. Here, we present evidence for the origin of H-2 metabolism genes in eukaryotes from an ancestor of the Anoxychlamydiales-a group of anaerobic chlamydiae, newly described here, from marine sediments. Among Chlamydiae, these bacteria uniquely encode genes for H-2 metabolism and other anaerobiosis-associated pathways. Phylogenetic analyses of several components of H-2 metabolism reveal that Anoxychlamydiales homologs are the closest relatives to eukaryotic sequences. We propose that an ancestor of the Anoxychlamydiales contributed these key genes during the evolution of eukaryotes, supporting a mosaic evolutionary origin of eukaryotic metabolism

Functional reconstruction of a eukaryotic-like E1/E2/(RING) E3 ubiquitylation cascade from an uncultured archaeon.

The covalent modification of protein substrates by ubiquitin regulates a diverse range of critical biological functions. Although it has been established that ubiquitin-like modifiers evolved from prokaryotic sulphur transfer proteins it is less clear how complex eukaryotic ubiquitylation system arose and diversified from these prokaryotic antecedents. The discovery of ubiquitin, E1-like, E2-like and small-RING finger (srfp) protein components in the Aigarchaeota and the Asgard archaea superphyla has provided a substantive step toward addressing this evolutionary question. Encoded in operons, these components are likely representative of the progenitor apparatus that founded the modern eukaryotic ubiquitin modification systems. Here we report that these proteins from the archaeon Candidatus 'Caldiarchaeum subterraneum' operate together as a bona fide ubiquitin modification system, mediating a sequential ubiquitylation cascade reminiscent of the eukaryotic process. Our observations support the hypothesis that complex eukaryotic ubiquitylation signalling pathways have developed from compact systems originally inherited from an archaeal ancestor

Binding and transport of D-aspartate by the glutamate transporter homologue GltTk

Mammalian glutamate transporters are crucial players in neuronal communication as they perform neurotransmitter reuptake from the synaptic cleft. Besides L-glutamate and L-aspartate, they also recognize D-aspartate, which might participate in mammalian neurotransmission and/or neuromodulation. Much of the mechanistic insight in glutamate transport comes from studies of the archaeal homologues GltPh from Pyrococcus horikoshii and GltTk from Thermococcus kodakarensis. Here, we show that GltTk transports D-aspartate with identical Na+ : substrate coupling stoichiometry as L-aspartate, and that the affinities (Kd and Km) for the two substrates are similar. We determined a crystal structure of GltTk with bound D-aspartate at 2.8 Å resolution. Comparison of the L- and D-aspartate bound GltTk structures revealed that D-aspartate is accommodated with only minor rearrangements in the structure of the binding site. The structure explains how the geometrically different molecules L- and D-aspartate are recognized and transported by the protein in the same way

Microbial community assembly and evolution in subseafloor sediment

Bacterial and archaeal communities inhabiting the subsurface seabed live under strong energy limitation and have growth rates that are orders of magnitude slower than laboratory-grown cultures. It is not understood how subsurface microbial communities are assembled and whether populations undergo adaptive evolution or accumulate mutations as a result of impaired DNA repair under such energy-limited conditions. Here we use amplicon sequencing to explore changes of microbial communities during burial and isolation from the surface to the > 5,000-y-old subsurface of marine sediment and identify a small core set of mostly uncultured bacteria and archaea that is present throughout the sediment column. These persisting populations constitute a small fraction of the entire community at the surface but become predominant in the subsurface. We followed patterns of genome diversity with depth in four dominant lineages of the persisting populations by mapping metagenomic sequence reads onto single-cell genomes. Nucleotide sequence diversity was uniformly low and did not change with age and depth of the sediment. Likewise, therewas no detectable change inmutation rates and efficacy of selection. Our results indicate that subsurface microbial communities predominantly assemble by selective survival of taxa able to persist under extreme energy limitation