Bell violation with entangled photons, free of the fair-sampling assumption

The violation of a Bell inequality is an experimental observation that forces one to abandon a local realistic worldview, namely, one in which physical properties are (probabilistically) defined prior to and independent of measurement and no physical influence can propagate faster than the speed of light. All such experimental violations require additional assumptions depending on their specific construction making them vulnerable to so-called "loopholes." Here, we use photons and high-efficiency superconducting detectors to violate a Bell inequality closing the fair-sampling loophole, i.e. without assuming that the sample of measured photons accurately represents the entire ensemble. Additionally, we demonstrate that our setup can realize one-sided device-independent quantum key distribution on both sides. This represents a significant advance relevant to both fundamental tests and promising quantum applications

From waste to health-supporting molecules: biosynthesis of natural products from lignin-, plastic- and seaweed-based monomers using metabolically engineered Streptomyces lividans

Background Transforming waste and nonfood materials into bulk biofuels and chemicals represents a major stride in creating a sustainable bioindustry to optimize the use of resources while reducing environmental footprint. However, despite these advancements, the production of high-value natural products often continues to depend on the use of frst-generation substrates, underscoring the intricate processes and specifc requirements of their bio‑ syntheses. This is also true for Streptomyces lividans, a renowned host organism celebrated for its capacity to produce a wide array of natural products, which is attributed to its genetic versatility and potent secondary metabolic activ‑ ity. Given this context, it becomes imperative to assess and optimize this microorganism for the synthesis of natural products specifcally from waste and nonfood substrates. Results We metabolically engineered S. lividans to heterologously produce the ribosomally synthesized and post‑ translationally modifed peptide bottromycin, as well as the polyketide pamamycin. The modifed strains success‑ fully produced these compounds using waste and nonfood model substrates such as protocatechuate (derived from lignin), 4-hydroxybenzoate (sourced from plastic waste), and mannitol (from seaweed). Comprehensive tran‑ scriptomic and metabolomic analyses ofered insights into how these substrates infuenced the cellular metabolism of S. lividans. In terms of production efciency, S. lividans showed remarkable tolerance, especially in a fed-batch process using a mineral medium containing the toxic aromatic 4-hydroxybenzoate, which led to enhanced and highly selective bottromycin production. Additionally, the strain generated a unique spectrum of pamamycins when cul‑ tured in mannitol-rich seaweed extract with no additional nutrients. Conclusion Our study showcases the successful production of high-value natural products based on the use of var‑ ied waste and nonfood raw materials, circumventing the reliance on costly, food-competing resources. S. lividans exhibited remarkable adaptability and resilience when grown on these diverse substrates. When cultured on aromatic compounds, it displayed a distinct array of intracellular CoA esters, presenting promising avenues for polyketide pro‑ duction. Future research could be focused on enhancing S. lividans substrate utilization pathways to process the intri‑ cate mixtures commonly found in waste and nonfood sources more efciently

Microparticles globally reprogram Streptomyces albus toward accelerated morphogenesis, streamlined carbon core metabolism, and enhanced production of the antituberculosis polyketide pamamycin

Streptomyces spp. are a rich source for natural products with recognized industrial value, explaining the high interest to improve and streamline the performance of in these microbes. Here, we studied the production of pamamycins, macrodiolide homologs with a high activity against multiresistant pathogenic microbes, using recombinant Streptomyces albus J1074/R2. Talc particles (hydrous magnesium silicate, 3MgO·4SiO2·H2O) of micrometer size, added to submerged cultures of the recombinant strain, tripled pamamycin production up to 50 mg/L. Furthermore, they strongly affected morphology, reduced the size of cell pellets formed by the filamentous microbe during the process up to sixfold, and shifted the pamamycin spectrum to larger derivatives. Integrated analysis of transcriptome and precursor (CoA thioester) supply of particle‐enhanced and control cultures provided detailed insights into the underlying molecular changes. The microparticles affected the expression of 3,341 genes (56% of all genes), revealing a global and fundamental impact on metabolism. Morphology‐associated genes, encoding major regulators such as SsgA, RelA, EshA, Factor C, as well as chaplins and rodlins, were found massively upregulated, indicating that the particles caused a substantially accelerated morphogenesis. In line, the pamamycin cluster was strongly upregulated (up to 1,024‐fold). Furthermore, the microparticles perturbed genes encoding for CoA‐ester metabolism, which were mainly activated. The altered expression resulted in changes in the availability of intracellular CoA‐esters, the building blocks of pamamycin. Notably, the ratio between methylmalonyl CoA and malonyl‐CoA was increased fourfold. Both metabolites compete for incorporation into pamamycin so that the altered availability explained the pronounced preference for larger derivatives in the microparticle‐enhanced process. The novel insights into the behavior of S. albus in response to talc appears of general relevance to further explore and upgrade the concept of microparticle enhanced cultivation, widely used for filamentous microbes

Systems biology of industrial oxytetracycline production in Streptomyces rimosus: the secrets of a mutagenized hyperproducer

Background Oxytetracycline which is derived from Streptomyces rimosus, inhibits a wide range of bacteria and is industrially important. The underlying biosynthetic processes are complex and hinder rational engineering, so industrial manufacturing currently relies on classical mutants for production. While the biochemistry underlying oxytetracycline synthesis is known to involve polyketide synthase, hyperproducing strains of S. rimosus have not been extensively studied, limiting our knowledge on fundamental mechanisms that drive production. Results In this study, a multiomics analysis of S. rimosus is performed and wild-type and hyperproducing strains are compared. Insights into the metabolic and regulatory networks driving oxytetracycline formation were obtained. The overproducer exhibited increased acetyl-CoA and malonyl CoA supply, upregulated oxytetracycline biosynthe‑ sis, reduced competing byproduct formation, and streamlined morphology. These features were used to synthesize bhimamycin, an antibiotic, and a novel microbial chassis strain was created. A cluster deletion derivative showed enhanced bhimamycin production. Conclusions This study suggests that the precursor supply should be globally increased to further increase the expression of the oxytetracycline cluster while maintaining the natural cluster sequence. The mutagenized hyper‑ producer S. rimosus HP126 exhibited numerous mutations, including large genomic rearrangements, due to natural genetic instability, and single nucleotide changes. More complex mutations were found than those typically observed in mutagenized bacteria, impacting gene expression, and complicating rational engineering. Overall, the approach revealed key traits infuencing oxytetracycline production in S. rimosus, suggesting that similar studies for other antibi‑ otics could uncover general mechanisms to improve production

The Role of Mobile Health Technologies in Allergy Care:an EAACI Position Paper

Mobile health (mHealth) uses mobile communication devices such as smartphones and tablet computers to support and improve health-related services, data and information flow, patient self-management, surveillance, and disease management from the moment of first diagnosis to an optimized treatment. The European Academy of Allergy and Clinical Immunology created a task force to assess the state of the art and future potential of mHealth in allergology. The task force endorsed the "Be He@lthy, Be Mobile" WHO initiative and debated the quality, usability, efficiency, advantages, limitations, and risks of mobile solutions for allergic diseases. The results are summarized in this position paper, analyzing also the regulatory background with regard to the "General Data Protection Regulation" and Medical Directives of the European Community. The task force assessed the design, user engagement, content, potential of inducing behavioral change, credibility/accountability, and privacy policies of mHealth products. The perspectives of healthcare professionals and allergic patients are discussed, underlining the need of thorough investigation for an effective design of mHealth technologies as auxiliary tools to improve quality of care. Within the context of precision medicine, these could facilitate the change in perspective from clinician- to patient-centered care. The current and future potential of mHealth is then examined for specific areas of allergology, including allergic rhinitis, aerobiology, allergen immunotherapy, asthma, dermatological diseases, food allergies, anaphylaxis, insect venom, and drug allergy. The impact of mobile technologies and associated big data sets are outlined. Facts and recommendations for future mHealth initiatives within EAACI are listed

Monthly variation in the probability of presence of adult Culicoides populations in nine European countries and the implications for targeted surveillance

Background: Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) are small hematophagous insects responsible for the transmission of bluetongue virus, Schmallenberg virus and African horse sickness virus to wild and domestic ruminants and equids. Outbreaks of these viruses have caused economic damage within the European Union. The spatio-temporal distribution of biting midges is a key factor in identifying areas with the potential for disease spread. The aim of this study was to identify and map areas of neglectable adult activity for each month in an average year. Average monthly risk maps can be used as a tool when allocating resources for surveillance and control programs within Europe. Methods : We modelled the occurrence of C. imicola and the Obsoletus and Pulicaris ensembles using existing entomological surveillance data from Spain, France, Germany, Switzerland, Austria, Denmark, Sweden, Norway and Poland. The monthly probability of each vector species and ensembles being present in Europe based on climatic and environmental input variables was estimated with the machine learning technique Random Forest. Subsequently, the monthly probability was classified into three classes: Absence, Presence and Uncertain status. These three classes are useful for mapping areas of no risk, areas of high-risk targeted for animal movement restrictions, and areas with an uncertain status that need active entomological surveillance to determine whether or not vectors are present. Results: The distribution of Culicoides species ensembles were in agreement with their previously reported distribution in Europe. The Random Forest models were very accurate in predicting the probability of presence for C. imicola (mean AUC = 0.95), less accurate for the Obsoletus ensemble (mean AUC = 0.84), while the lowest accuracy was found for the Pulicaris ensemble (mean AUC = 0.71). The most important environmental variables in the models were related to temperature and precipitation for all three groups. Conclusions: The duration periods with low or null adult activity can be derived from the associated monthly distribution maps, and it was also possible to identify and map areas with uncertain predictions. In the absence of ongoing vector surveillance, these maps can be used by veterinary authorities to classify areas as likely vector-free or as likely risk areas from southern Spain to northern Sweden with acceptable precision. The maps can also focus costly entomological surveillance to seasons and areas where the predictions and vector-free status remain uncertain

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Auswirkungen der extensiven Bewirtschaftung von Dauergrünland auf Heuschrecken und Hummeln

Das Land Nordrhein-Westfalen fördert im Zuge des "NRW-Programm Ländlicher Raum 2007-2013" Maßnahmen zur Erhaltung und zur umweltverträglichen Nutzung des Dauergrünlandes. Hierzu zählt die Agrarumweltmaßnahme "Extensive Dauergrünlandnutzung", deren Umsetzungsschwerpunkte sich in den Mittelgebirgslagen befinden. Während die günstige Wirkung dieser Maßnahme auf den abiotischen Ressourcenschutz anerkannt ist, liegen bislang nur wenige Erkenntnisse zu möglichen positiven Effekten auf die Biodiversität vor. Ziel der vorliegenden Studie war es, am Beispiel von Heuschrecken und Hummeln die Auswirkungen der Agrarumweltmaßnahme in Nordrhein-Westfalen zu analysieren. Die Untersuchungen wurden in den Jahren 2013 bis 2015 in den Mittelgebirgsregionen der Eifel und des Süderberglandes durchgeführt. Es erfolgte ein Vergleich von Grünlandflächen, die nach den Vorgaben der Agrarumweltmaßnahme bewirtschaftet wurden (AUM-Flächen) mit benachbarten Grünlandflächen ohne Bewirtschaftungsauflagen (VGL-Flächen). In der Eifel wurde als Referenz zusätzlich Grünland einbezogen, das nach Auflagen des Vertragsnaturschutzes bewirtschaftet wurde (VNS-Flächen). In den Jahren 2013 und 2014 wurden 24 Grünlandflächen (10 AUM, 10 VGL, 4 VNS) und in 2015 insgesamt 35 Grünlandflächen (14 AUM, 14 VGL, 7 VNS) untersucht. In den drei Untersuchungsjahren wurden von Mai bis September Heuschrecken per Streifnetzmethode und Hummeln per Sichtfang entlang festgelegter Transekte erfasst. Ein qualitativer Nachweis der Heuschreckenarten erfolgte zusätzlich anhand akustischer Merkmale. Auf allen Untersuchungsflächen wurden melittophile Blütenpflanzen und deren Dichte sowie die Vegetationshöhe der Pflanzenbestände erfasst. Ergänzend wurden innerhalb eines Landschaftsausschnitts mit einem Radius von einem Kilometer um die Untersuchungsflächen Biotop- und Nutzungstypen kartiert. Insgesamt wurden auf den Grünlandflächen in der Eifel und im Süderbergland 19 Heuschreckenarten nachgewiesen. Die Artenzusammensetzung der Heuschreckengemeinschaften beider Landschaftsräume war ähnlich. Auf den AUM-Flächen wurden im Vergleich zu den VGL-Flächen in der Tendenz eine höhere Artenzahl und Diversität von Heuschrecken festgestellt. VNS-Flächen wiesen gegenüber den VGL-Flächen signifikant höhere Werte der ermittelten Diversitätsparameter auf. Bei der Zusammensetzung der unterschiedlichen ökologischen Gruppen gab es zwischen den Bewirtschaftungsvarianten keine signifikanten Unterschiede. Der Anteil von Flächen, mit Vorkommen von Langfühlerschrecken, war bei den AUM-Flächen höher als bei den VGL-Flä-chen. Der verstärkte Nachweis von Larvenstadien belegt deren bessere Eignung als Reproduktionshabitat. Auf den VNS-Flächen traten Langfühlerschrecken mit der höchsten Stetigkeit auf. Das Auftreten von Langfühlerschrecken trägt maßgeblich zu der Diversität der Heuschreckengemeinschaften bei. Die Abundanz der Langfühlerschreckenarten korrelierte mit der Vegetationshöhe der Untersuchungsflächen. Die Hummelgemeinschaften umfassten insgesamt 16 Arten. Im Süderbergland (8 Hum-melarten) wurde gegenüber der Eifel (15 Hummelarten) eine deutlich geringere Artendiversität erfasst. Die Zusammensetzung der ökologischen Gruppen unterschied sich stärker zwischen den Landschaftsräumen als zwischen den Bewirtschaftungsvarianten. Die Artenzahlen, Abundanzen und Diversitätswerte der Hummelgemeinschaften waren auf den AUM-Flächen tendenziell höher als auf den VGL-Flächen. Auf den VNS-Flächen wurden gegenüber den VGL-Flächen signifikant höhere Werte dieser Diversitätsparameter festgestellt. Die höheren Individuenzahlen auf den AUM-Flächen resultieren insbesondere aus der hohen Abundanz der Distanzsammler Bombus lucorum und Bombus lapidarius. Distanzsammler sind in der Lage, weite Strecken zwi-schen ihrem Nist- und Nahrungshabitat zurückzulegen. Ein entscheidender Faktor für ihre Abundanz ist die Blütendichte. Zum Zeitpunkt der dominanten Kleeblüte im Juni nutzen sie die AUM-Flächen als Sammelhabitate. Nahbereichssammler wurden mit höheren Artenzahlen auf den AUM- und VNS-Flächen nachgewiesen. Nahbereichssamller sind auf ein stetes Blütenangebot in der direkten Umgebung ihres Nistplatzes angewiesen. Da die meisten Hummelarten Generalisten sind, wirkte sich die Vielfalt von Blütenpflanzenarten hingegen nur in geringem Umfang auf die Hummeldiversität aus. Die Ergebnisse zeigen, dass Heuschrecken und Hummeln von extensiven Grünlandbewirtschaftungsformen profitieren. Die verpflichtende reduzierte Düngung und die Festlegung einer maximalen Viehesatzstärke im Betrieb von 1,4 raufutterfressenden Großvieheinheiten (RGV) je Hektar Hauptfutterfläche bei der Agrarumweltmaßnahme „Extensive Dauergrünlandnutzung“ sind zweifelsfrei wichtige Ansatzpunkte zur Förderung der Biodiversität und Stabilisierung von Graslandökosystemen. Mit der Reduzierung des Düngeniveaus wird auf vielen Standorten erst nach vielen Jahren ein Anstieg der Artenvielfalt des Pflanzenbestandes erreicht. Trotzdem ist davon auszugehen, dass durch die generelle Ausdehnung des Flächenumfangs der Agrarumweltmaßnahme „Extensive Dauergrünlandnutzung“ positive Effekte auf wirbellose Tiere erzielt werden. Die Ergebnisse der VNS-Flächen zeigen, dass die Reduzierung der Schnitttermine ein wichtiger Ansatzpunkt für die Steigerung der Habitatqualität von Grünlandflächen für Heuschrecken und Hummeln ist. Die Anzahl der Grünlandschnitte beeinflusst sowohl die zeitliche Verfügbarkeit des Blütenangebots für Hummeln als auch die Verfügbarkeit von Reproduktionsräumen für Heuschrecken. Die positive Wirkung der Agrarumweltmaßnahme „Extensive Dauergrünlandnutzung“ kann folglich durch die Festlegung von „Ruhephasen“ (ohne Mahd oder Beweidung) oder „Nutzungsverzichtsstreifen“ (s.u.) weiter gestärkt werden. Bei der Ausrichtung dieser zusätzlichen Maßnahmen müssen allerdings sowohl die Anforderungen von Milchvieh- und Mutterkuhbetrieben an die Futterleistung und -qualität des Grünlandes als auch die generelle Verwertbarkeit des Aufwuchses beachtet werden. Ein Hochschnitt (mindestens 8 cm oberhalb des Bodens) könnte dazu beitragen, niedrigwüchsige Kleearten und andere Blütenpflanzen als Nahrungsressource für Hummeln auch nach der Mahd zu erhalten und darüber hinaus die Mortalitätsrate der Heuschrecken durch mechanische Einwirkung bei der Mahd zu reduzieren. Als Zusatzmaßnahme für die naturschutzgerechte Grünlandbewirtschaftung schlagen wir vor, nach der ersten Nutzung etwa 5 % der Fläche (als Streifen oder Insel) bis ins Folgejahr von der weiteren Bewirtschaftung auszusparen. Folgende Effekte werden erwartet: • Erhöhung der Strukturvielfalt - Bereitstellung zusätzlicher Habitate durch dauerhafte Strukturen innerhalb der Landschaft auch im Winter, • Erhaltung von Rückzugsräumen - Quellbiotop für die Wiederbesiedlung der Flä-che nach der Nutzung, • Erhaltung von Reproduktionshabitaten - insbesondere für pflanzenbrütende Heuschreckenarten, • Erhaltung von Nahrungshabitaten für Hummeln - Gewährleistung eines kontinuierlichen Ressourcenangebotes, • Vernetzung - Vor allem eine streifenförmige Anordnung dient als Ausbreitungsachse und kann so Lebensräume miteinander verbinden.The state of North Rhine-Westphalia is promoting measures for the conservation and sustainable use of permanent grassland as part of the "NRW Rural Development Pro-gram 2007 - 2013". This includes the agri-environmental measure "Extensive Grass-land Use", which focus is in the low mountain range. While the beneficial effects of this measure on abiotic resource conservation have been recognized, only a few findings on possible positive effects on biodiversity are available. The aim of the present study was to analyze the effects of the agri-environmental measure on invertebrates in North Rhine-Westphalia using the example of grasshoppers and bumblebees. Studies were conducted from 2013 to 2015 in the low mountain regions of the Eifel and the Süderbergland (North Rhine-Westphalia, Germany). A comparison was made be-tween grassland plots, which were managed according to the requirements of the agri-environment measure (AUM), and neighboring grassland sites without management obligations (VGL). As a reference, we additionally investigated grassland, which was managed according to the provisions of contractual nature conservation (VNS) in half of the areas. In 2013 and 2014, twenty four grassland sites (10 AUM, 10 VGL, 4 VNS) were examined, and in 2015, thirty five sites (14 AUM, 14 VGL, 7 VNS) were examined. Over the three years of investigation, grasshoppers and bumble bees were recorded from May to September, captured using an insect net along fixed transects. A qualita-tive proof of grasshoppers was also made by means of acoustic features. Melittophilic flowering plants and their density, as well as the vegetation height of the grasslands, were recorded on all investigation sites. In addition, biotope and land use types were mapped within a landscape section with a radius of one kilometer around the investi-gation sites. A total of 19 grasshopper species were found on the grasslands in the Eifel and the Süderbergland. The species composition of the grasshopper communities was similar between both regions. The AUM sites trend towards higher values of spe-cies number and species diversity of grasshoppers compared to the VGL sites. Signif-icantly, higher values of these diversity parameters were found on the VNS sites com-pared to the VGL sites. There were no major differences in the composition of different ecological groups across the management schemes. The share of sites, were Ensifera occured, was higher among the AUM sites than among the VGL sites. The more fre-quent detection of larval stages demonstrates their superior suitability as a reproduc-tion habitat. With the highest continuity, Ensifera occurred on the VNS sites. The oc-currence of Ensifera contributes significantly to the diversity of grasshopper communi-ties. Most Ensifera species are plant breeders and need high-growing vegetation for reproduction. Their abundance correlated with the vegetation height of the examination sites. The bumblebee communities included 16 species. In the Süderbergland (8 bumblebee species) an obvious lower species number was recorded towards the Eifel (15 bum-blebee species). The composition of the ecological groups differed more strongly be-tween the two regions than between the different management schemes. The species numbers, abundances, and diversity values of the bumblebee communities were higher on the AUM sites than on the VGL sites. On the VNS sites significantly higher values of these diversity parameters were found versus the VGL sites. The higher in-dividual numbers on the AUM sites are a result of the high abundance of the long-distance foraging species Bombus lucorum and Bombus lapidarius. These species are able to cover far distances between their nesting and foraging habitats. A decisive factor for their abundance is the flower density. At the time of the dominant clover blossom in June, they used the AUM sites for foraging. Short distance foragers, were detected with higher numbers of species on the AUM and VNS sites. These species depend on a constant supply of flowers in the immediate vicinity of their nesting sites. Since most bumblebee species are generalists, the variety of flowering species has only a small influence on their diversity. The results show that grasshoppers and bumblebees benefit from extensive grassland management. The compulsory reduced fertilization and the establishment of a maxi-mum stocking of 1.4 RGV in the agri-environmental measure are undoubtedly im-portant starting points for the promotion of biodiversity and stabilization of grassland ecosystems. By reduction of the level of fertilisation, on many sites the number of dif-ferent plant species is increasing only after many years. Nevertheless, it is to be as-sumed that positive effects on invertebrate animals are achieved by the general exten-sion of the area of the agri-environmental measure "Extensive Grassland Use". The results of the VNS reference sites show that the reduction of mowing frequency is an important starting point for the increase of the habitat quality of grassland for grass-hoppers and bumblebees. The number of cuts affects both the temporal availability of the flower resources for bumblebees as well as the availability of reproduction sites for grasshoppers. Therefore, the positive effect of the agri-environmental measure can be strengthened by the definition of "resting phases" (without mowing or grazing) or "old gras strips". However, when enacting these additional measures, the demands of dairy farms for silage quality, as well as the general usability of the yield, have to be consid-ered. A high-cut (at least 8 cm above the ground) could help to maintain low-growing clover species and other flowering plants as a feeding resource for bumblebees even after mowing. Furthermore, it might reduce the mortality rate of grasshoppers by me-chanical means during mowing. As an additional measure for grassland management appropriate to nature conservation, we propose to save about 5% of the area (as a strip or island) from further cultivation, after the first cut until the following year. We expect the following positive effects: - Increasing structural diversity - providing additional habitats through permanent structures within the landscape, even in winter, • Preservation of retreats - source biotope for the resettlement of the site after mowing, • Preservation of reproduction habitats - especially for plant breeding grasshop-per species, • Preservation of foraging resources for bumblebees - Ensuring a continuous sup-ply of resources, • Connectivity - Especially a strip-shaped arrangement serves as an axis for dis-persal and can thus connect habitats

Microparticles enhance the formation of seven major classes of natural products in native and metabolically engineered actinobacteria through accelerated morphological development

Kuhl M, Rückert C, Glaser L, et al. Microparticles enhance the formation of seven major classes of natural products in native and metabolically engineered actinobacteria through accelerated morphological development. Biotechnology and bioengineering. Accepted.Actinobacteria provide a rich spectrum of bioactive natural products and therefore display an invaluable source towards commercially valuable pharmaceuticals and agrochemicals. Here, we studied the use of inorganic talc microparticles (hydrous magnesium silicate, 3MgO·4SiO2·H2O, 10 m) as a general supplement to enhance natural product formation in this important class of bacteria. Added to cultures of recombinant Streptomyces lividans, talc enhanced production of the macrocyclic peptide antibiotic bottromycin A2 and its methylated derivative Met-bottromycin A2 up to 109 mg L-1 , the highest titer reported so far. Hereby, the microparticles fundamentally affected metabolism. With 10 g L-1 talc, S. lividans grew to 40% smaller pellets and, using RNA sequencing, revealed accelerated morphogenesis and aging, indicated by early upregulation of developmental regulator genes such as ssgA, ssgB, wblA, sigN and bldN. Furthermore, the microparticles re-balanced the expression of individual bottromycin cluster genes, resulting in a higher macrocyclization efficiency at the level of BotAH and correspondingly lower levels of non-cyclized shunt by-products, driving the production of mature bottromycin. Testing a variety of Streptomyces species, talc addition resulted in up to 13-fold higher titers for the RiPPs bottromycin and cinnamycin, the alkaloid undecylprodigiosin, the polyketide pamamycin, the tetracycline-type oxytetracycline, and the anthramycin-analogues usabamycins. Moreover, talc addition boosted production in other actinobacteria, outside of the genus of Streptomyces: vancomycin (Amycolatopsis japonicum DSM 44213), teicoplanin (Actinoplanes teichomyceticus ATCC 31121), and the angucyclinone-type antibiotic simocyclinone (Kitasatospora sp.). For teicoplanin, the microparticles were even crucial to activate production. Taken together, the use of talc was beneficial in 75% of all tested cases and optimized natural and heterologous hosts forming the substance of interest with clusters under native and synthetic control. Given its simplicity and broad benefits, microparticle-supplementation appears as an enabling technology in natural product research of these most important microbes. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved

From waste to health-supporting molecules: biosynthesis of natural products from lignin-, plastic- and seaweed-based monomers using metabolically engineered Streptomyces lividans

Abstract Background Transforming waste and nonfood materials into bulk biofuels and chemicals represents a major stride in creating a sustainable bioindustry to optimize the use of resources while reducing environmental footprint. However, despite these advancements, the production of high-value natural products often continues to depend on the use of first-generation substrates, underscoring the intricate processes and specific requirements of their biosyntheses. This is also true for Streptomyces lividans, a renowned host organism celebrated for its capacity to produce a wide array of natural products, which is attributed to its genetic versatility and potent secondary metabolic activity. Given this context, it becomes imperative to assess and optimize this microorganism for the synthesis of natural products specifically from waste and nonfood substrates. Results We metabolically engineered S. lividans to heterologously produce the ribosomally synthesized and posttranslationally modified peptide bottromycin, as well as the polyketide pamamycin. The modified strains successfully produced these compounds using waste and nonfood model substrates such as protocatechuate (derived from lignin), 4-hydroxybenzoate (sourced from plastic waste), and mannitol (from seaweed). Comprehensive transcriptomic and metabolomic analyses offered insights into how these substrates influenced the cellular metabolism of S. lividans. In terms of production efficiency, S. lividans showed remarkable tolerance, especially in a fed-batch process using a mineral medium containing the toxic aromatic 4-hydroxybenzoate, which led to enhanced and highly selective bottromycin production. Additionally, the strain generated a unique spectrum of pamamycins when cultured in mannitol-rich seaweed extract with no additional nutrients. Conclusion Our study showcases the successful production of high-value natural products based on the use of varied waste and nonfood raw materials, circumventing the reliance on costly, food-competing resources. S. lividans exhibited remarkable adaptability and resilience when grown on these diverse substrates. When cultured on aromatic compounds, it displayed a distinct array of intracellular CoA esters, presenting promising avenues for polyketide production. Future research could be focused on enhancing S. lividans substrate utilization pathways to process the intricate mixtures commonly found in waste and nonfood sources more efficiently