Natural Substrate Research Articles

Isolation and comparative analysis of culturable bacterial communities associated with life stages, breeding and rearing substrates of Culicoides oxystoma Kieffer (Diptera: Ceratopogonidae) vector of bluetongue virus.

Culicoides oxystoma Kieffer (Diptera: Ceratopogonidae) has been vectoring several arboviruses, protozoa and nematodes, leading to mortality and morbidity of livestock and wild ruminants in the tropics and subtropics. Insight into the bacterial communities associated with the vector species must be worked out. This work tries to inventorize the bacterial communities associated with this important vector species. Acquisition of gut microbiota may be the parental origin, while some are obtained through feeding during larval stages. Culicoides oxystoma possesses semi-aquatic life cycle strategies for egg-laying and larval survival. The bacteria associated with C. oxystoma were compared throughout (i) life stages: egg, larval instars, pupa, adult: male and female obtained from laboratory colony; (ii) field-collected adult: male and age-graded females; and (iii) natural breeding substrate and artificial rearing substrate. The culture-dependent bacteria were identified by Sanger sequencing of 16S rRNA, and haemolytic bacteria were screened on blood agar. Results show that Firmicutes and Proteobacteria are the predominant Phyla, of which Bacillus spp. was the most abundant across the life stages. Across the life history, Bacillus cereus, Bacillus pumilus, Bacillus tropicus, Lysinibacillus sp. and Paenibacillus sp. were retrieved routinely. Bacillus cereus and Alcaligenes faecalis were detected in the lab-reared specimens and shared between the natural breeding site and rearing medium. From the adults trapped across two locations, B. cereus, Bacillus flexus, A. faecalis, Enterococcus faecium and Pseudomonas sp. were isolated. The bacterial species associated with this vector may influence various physiological traits, such as vectorial capacity, digestion and larval development, which need further investigation.

Impacts of Dislocations and Residual Thermal Tension on Monolithically Integrated InGaP/GaAs/Si Triple‐Junction Solar Cells

Direct epitaxy of III−V materials on Si is a promising approach for highly stable, scalable, and efficient Si‐based multijunction solar cells. However, challenges lie in overcoming epitaxial dislocations and residual thermal strain generated by lattice constant and thermal‐expansion‐coefficient mismatches, respectively. Herein, a 15.2% efficient InGaP/GaAs/Si triple‐junction solar cell with an open‐circuit voltage of 2.36 V by using In0.10Al0.16Ga0.74As digital‐alloy dislocation filter layers is first demonstrated. The filter layers are utilized in the n‐GaAs buffer on Si to reduce threading dislocation density to 4 × 107 cm−2 while maintaining optical transparency to Si bottom cell. Then, the impacts of threading dislocations and residual tension on InGaP/GaAs/Si cells are systematically investigated by comparing them to the co‐grown InGaP/GaAs tandem cells on a native GaAs substrate. Based on the comparative analysis, a strategy to suppress material deformation and defect formation toward 30% efficient InGaP/GaAs/Si triple‐junction solar cells is proposed.

Production, purification and characterization of phytase from Pichia kudriavevii FSMP-Y17and its application in layers feed.

Phytase, recognized for its ability to enhance the nutritional value of phytate-rich foods, has has gained significant prominence. The production of this enzyme has been significantly boosted while preserving economic efficiency by utilizing natural substrates and optimizing essential factors. This study focuses on optimizing phytase production through solid-state fermentation and evaluating its effectiveness in enhancing nutrient utilization in chicken diets. The objective is to optimize phytase production via solid-state fermentation, characterize purified phytase properties, and assess its impact on nutrient utilization in chicken diets. Through these objectives, we aim to deepen understanding of phytase's role in poultry nutrition and contribute to more efficient feed formulations for improved agricultural outcomes. We utilized solid-state fermentation with Pichia kudriavzevii FSMP-Y17 yeast on orange peel substrate, optimizing variables like temperature, pH, incubation time, and supplementing with glucose and ammonium sulfate. Following fermentation, we purified the phytase enzyme using standard techniques, characterizing its properties, including molecular weight, optimal temperature and pH, substrate affinity, and kinetic parameters. The optimized conditions yielded a remarkable phytase yield of 7.0 U/gds. Following purification, the enzyme exhibited a molecular weight of 64kDa and displayed optimal activity at 55°C and pH 5.5, with kinetic parameters (Km = 3.39 × 10-3M and a Vmax of 7.092mM/min) indicating efficient substrate affinity. The addition of purified phytase to chicken diets resulted in significant improvements in nutrient utilization and overall performance, including increased feed intake, improved feed conversion ratio, enhanced bird growth, better phosphorus retention, and improved egg production and quality. By addressing challenges associated with phytate-rich diets, such as reduced nutrient availability and environmental pollution, phytase utilization promotes animal welfare and sustainability in poultry production.

Hematoxylin, an Alternative Substrate of Tyrosinase.

Mushroom tyrosinase from Agaricus bisporus (abTYR) is often used during the development of tyrosinase inhibitors for medicinal and cosmetic purposes. In the search for novel tyrosinase inhibitors, this study identified hematoxylin as an alternative substrate for abTYR. The interaction of hematoxylin with abTYR was investigated through spectrophotometric and chromatographic analyses. The results showed that hematoxylin acted as an abTYR substrate and exhibited Michaelis-Menten kinetic behaviour at concentrations below 1.25 mM. The substrate properties of hematoxylin were similar to the natural tyrosinase substrate, L-3,4-dihydroxyphenylalanine (L-DOPA), with regards to Km, while Vmax was eightfold lower. The main oxidation product formed during the reaction of abTYR with hematoxylin was identified as hematein. This is the first report of the interaction of hematoxylin with abTYR.

Extracellular Vesicles of the Probiotic Escherichia coli Nissle 1917 Reduce PepT1 Levels in IL-1β-Treated Caco-2 Cells via Upregulation of miR-193a-3p.

PepT1, a proton-coupled oligopeptide transporter, is crucial for intestinal homeostasis. It is mainly expressed in small intestine enterocytes, facilitating the absorption of di/tri-peptides from dietary proteins. In the colon, PepT1 expression is minimal to prevent excessive responses to proinflammatory peptides from the gut microbiota. However, increased colonic PepT1 is linked to chronic inflammatory diseases and colitis-associated cancer. Despite promising results from animal studies on the benefits of extracellular vesicles (EVs) from beneficial gut commensals in treating IBD, applying probiotic EVs as a postbiotic strategy in humans requires a thorough understanding of their mechanisms. Here, we investigate the potential of EVs of the probiotic Nissle 1917 (EcN) and the commensal EcoR12 in preventing altered PepT1 expression under inflammatory conditions, using an interleukin (IL)-1-induced inflammation model in Caco-2 cells. The effects are evaluated by analyzing the expression of PepT1 (mRNA and protein) and miR-193a-3p and miR-92b, which regulate, respectively, PepT1 mRNA translation and degradation. The influence of microbiota EVs on PepT1 expression is also analyzed in the presence of bacterial peptides that are natural substrates of colonic PepT1 to clarify how the regulatory mechanisms function under both physiological and pathological conditions. The main finding is that EcN EVs significantly decreases PepT1 protein via upregulation of miR-193a-3p. Importantly, this regulatory effect is strain-specific and only activates in cells exposed to IL-1β, suggesting that EcN EVs does not control PepT1 expression under basal conditions but can play a pivotal role in response to inflammation as a stressor. By this mechanism, EcN EVs may reduce inflammation in response to microbiota in chronic intestinal disorders by limiting the uptake of bacterial proinflammatory peptides.

Impact of microplastics on microbial community structure in the Qiantang river: A potential source of N2O emissions

This study aimed to investigate the spatial distribution of microplastics (MPs) and the features of the bacterial community in the Qiantang River urban river. Surface water samples from the Qiantang River were analyzed for this purpose. The results of the 16S high-throughput sequencing indicated that the microbial community diversity of MPs was significantly lower than in natural water but higher than in natural substrates. The biofilm of MPs was mainly composed of Enterobacteriaceae (28.00%), Bacillaceae (16.25%), and Phormidiaceae (6.75%). The biodiversity on MPs, natural water, and natural substrates varied significantly and was influenced by seasonal factors. In addition, the presence of MPs hindered the denitrification process in the aquatic environment and intensified N2O emission when the nitrate concentration was higher than normal. In particular, polyethylene terephthalate (PET) exhibited a 12% residue of NO3−-N and a 4.2% accumulation of N2O after a duration of 48 h. Further findings on gene abundance and cell viability provided further confirmation that PET had a considerable impact on reducing the expression of nirS (by 0.34-fold) and nosZ (by 0.53-fold), hence impeding the generation of nicotinamide adenine dinucleotide (NADH) (by 0.79-fold). Notably, all MPs demonstrated higher the nirK gene abundances than the nirS gene, which could account for the significant accumulation of N2O. The results suggest that MPs can serve as a novel carrier substrate for microbial communities and as a potential promoter of N2O emission in aquatic environments.

Electrochemical α‐Functionalization of N‐Aryl‐Activated Tertiary Amines

AbstractNumerous appropriately substituted pyridyl or phenyl groups serve as a particularly advantageous activation motif for the electrochemical oxidation of amines. Such groups enable a general, mild method for the electrochemical α‐functionalization of tertiary amines across numerous activating groups and amine scaffolds. Notably, the method accommodates an unprecedented range of nucleophile classes, allowing for the introduction of diverse functional groups to the readily prepared amine substrates. The utility of this method is then demonstrated through applications to unsymmetrical bisfunctionalization, site‐selective functionalization of N‐pyridyl amines vs. other activated amines, a formal synthesis of ivosidenib and the diversification of FDA‐approved drugs or natural product substrates.

Synthesis of cobalt- ferrite and zinc oxide metal nanoparticles based-bentonite using SDS and their investigation as catalysts in synthesis of benzylbarbiturocoumarins.

In this research, a suitable and efficient CoFe2O4@ZnO@Bentonite nano-catalyst was designed and synthesized by using zinc oxide (ZnO) and cobalt ferrite (CoFe2O4) nanoparticles and bentonite by microwave irradiation. Characteristics of the synthesized nanocomposite were investigated by Fourier transform infrared (FT-IR), scanning electron microscope (SEM), energy dispersive X-ray (EDX), transmission electron microscope (TEM), X-ray diffraction (XRD), Bruner- Emmett-Teller (BET) and vibrating sample magnetometer (VSM) techniques. The produced catalyst was effectively employed as a supported solid acid catalyst in mildly agitated three-component reactions involving aromatic aldehydes, 4-hydroxycoumarin, and 1,3-dimethyl-barbituric acid in a single pot to produce benzylbarbiturocoumarins. Starting materials were condensed via three C-C bond formation by CoFe2O4@ZnO@Bentonite as an efficient, recyclable, and environmentally safe nanocatalyst to obtain target products. The advantages of this method include using a natural substrate, small amounts of catalyst, aqueous media, performing reactions at ambient temperature, simple separation and purification of products, and good yields with short reaction times.

(Electronics and Photonics Division Award) Research Advances Wide- and Ultrawide-Bandgap Power Switching Devices

Wide bandgap semiconductors such as SiC and GaN (WBG) and emerging ultrawide-bandgap materials such as Ga2O3, AlGaN, and Diamond (UWBG) represent the next-generation materials for high performance medium voltage and high voltage power switch technology. Such devices have a wide range of immediate Naval applications, including high-power satellite communications and radar, unmanned underwater and aerial vehicles, ship drive components, and hybrid vehicle inverters. Vertical SiC power device technology has matured rapidly over the past two decades, owing to advances in substrates, a fundamental understanding of epitaxial growth to eliminate performance-limiting defects, as well as device design breakthroughs. This has enabled breakthroughs in highly integrated module design for medium voltage power conversion with switching frequency >100 kHz. In parallel, lateral GaN-based high electron mobility transistor (HEMT) technology has been highly successful for RF power amplifiers and is well positioned to supersede GaAs-based microwave circuits. Recently, GaN-based vertical and lateral power devices have attracted significant interest due to promising device results coupled with progress in native substrate, epitaxial growth, and processing technology developments. This seminar will present an overview of the WBG/UWBG power device efforts at NRL. This research has four primary focus areas – 1) thermal management in III-N power device structures by diamond thin film integration, 2) characterization of substrate materials and homoepitaxial layers used for drift regions of power devices identify appropriate specifications and benchmark performance, 3) process module development, particularly for selective area doping by ion implantation, and 4) pilot production manufacturing of 1.2kV-class PiN diodes including reliability assessments to identify and mitigate performance limiting defects.

Tyrosinase Inhibition and Antimelanogenic Effects of Resorcinol-Containing Compounds.

Tyrosinases (TYRs) are copper-containing metalloenzymes present in a large diversity of species. In human, hTYR is responsible for pivotal steps in melanogenesis, catalysing the oxidation of l-tyrosine to l-DOPA and further to dopaquinone. While numerous TYR inhibitors have been reported, polyphenolic compounds tend to dominate the literature. However, many of these compounds, particularly monophenols and catechols, have been identified as alternative substrates rather than true inhibitors, given their structural similarity to natural substrates. Resorcinol-containing compounds have emerged as promising candidates to address this challenge, as the meta-dihydroxy moiety in resorcinol demonstrates resistance to TYR-mediated oxidation, while retaining the favourable interactions with copper ions provided by the hydroxy groups. Although their precise mechanism of action remains debated, resorcinol derivatives have yielded some of the most active compounds against isolated mushroom and human TYRs, as well as clinically used dermocosmetic agents like rucinol and thiamidol, which exhibited very promising effects in patients with facial melasma. This review outlines the development of resorcinol-containing TYR inhibitors, categorized by scaffold type, ranging from simple alkyl analogues to intricate synthetic derivatives. Mechanistic insights about the resorcinol-TYR interaction are also presented and debated.

Unveiling the kinetic versatility of aryl-alcohol oxidases with different electron acceptors.

Introduction: Aryl-alcohol oxidase (AAO) shows a pronounced duality as oxidase and dehydrogenase similar to that described for other glucose-methanol-choline (GMC) oxidase/dehydrogenase superfamily proteins involved in lignocellulose decomposition. In this work, we detail the overall mechanism of AAOs from Pleurotus eryngii and Bjerkandera adusta for catalyzing the oxidation of natural aryl-alcohol substrates using either oxygen or quinones as electron acceptors and describe the crystallographic structure of AAO from B. adusta in complex with a product analogue. Methods: Kinetic studies with 4-methoxybenzyl and 3-chloro-4- methoxybenzyl alcohols, including both transient-state and steady-state analyses, along with interaction studies, provide insight into the oxidase and dehydrogenase mechanisms of these enzymes. Moreover, the resolution of the crystal structure of AAO from B. adusta allowed us to compare their overall folding and the structure of the active sites of both AAOs in relation to their activities. Results and Discussion: Although both enzymes show similar mechanistic properties, notable differences are highlighted in this study. In B. adusta, the AAO oxidase activity is limited by the reoxidation of the flavin, while in P. eryngii the slower step takes place during the reductive half-reaction, which determines the overall reaction rate. By contrast, dehydrogenase activity in both enzymes, irrespective of the alcohol participating in the reaction, is limited by the hydroquinone release from the active site. Despite these differences, both AAOs are more efficient as dehydrogenases, supporting the physiological role of this activity in lignocellulosic decay. This dual activity would allow these enzymes to adapt to different environments based on the available electron acceptors.

Engineering Living Material for Controlled Fragrance Release Utilizing Kluyveromyces marxianus CBS6556 and Adaptive Hydrogel.

The demand for controllable fragrance materials is substantial owing to their potential to impart enduring scents in a variety of applications. However, the practical application of such materials has been limited by challenges in tunable morphogenesis, structural variability, and adaptability to diverse conditions. In our study, we introduce a hybrid living material that integrates a genetically engineered strain of Kluyveromyces marxianus CBS6556 with an adaptive hydrogel. The engineered K. marxianus achieved temperature stability in 2-phenylethanol (2-PE) and 2-phenylethyl acetate (2-PEAc) production by expressing relevant genes in the 2-PE metabolic pathway using the high-temperature preferential promoter SSE1. The enhanced water retention capacity supports the metabolic activities of the encapsulated yeast cells, ensuring their survival and functionality over an extended period. Fragrance-releasing living material (FLM) is designed to controllably emit fragrance 2-PE by adjusting the microbial concentration within the hydrogel matrix. The FLM exhibits versatile adhesion capabilities, effectively binding to a spectrum of surfaces such as wood, textiles, and glass as well as to natural substrates like leaves. This adaptability enhances the material's applicability across various settings. Furthermore, FLM can be crafted into various forms, including microbeads, fibers, and films. This research opens up new horizons for controlled fragrance release of living materials.

Identification of a novel caspase cleavage motif AEAD

Infections of many viruses induce caspase activation to regulate multiple cellular pathways, including programmed cell death, immune signaling and etc. Characterizations of caspase cleavage sites and substrates are important for understanding the regulation mechanisms of caspase activation. Here, we identified and analyzed a novel caspase cleavage motif AEAD, and confirmed its caspase dependent cleavage activity in natural substrate, such as nitric oxide-associated protein 1 (NOA1). Fusing the enhanced green fluorescent protein (EGFP) with the mitochondrial marker protein Tom20 through the AEAD motif peptide localized EGFP to the mitochondria. Upon the activation of caspase triggered by Sendai virus (SeV) or herpes simplex virus type 1 (HSV-1) infection, EGFP diffusely localized to the cell due to the caspase-mediated cleavage, thus allowing visual detection of the virus-induced caspase activation. An AEAD peptide-derived inhibitor Z-AEAD-FMK were developed, which significantly inhibited the activities of caspases-1, -3, -6, -7, -8 and -9, exhibiting a broad caspase inhibition effect. The inhibitor further prevented caspases-mediated cleavage of downstream substrates, including BID, PARP1, LMNA, pro-IL-1β, pro-IL-18, GSDMD and GSDME, protecting cells from virus-induced apoptotic and pyroptotic cell death. Together, our findings provide a new perspective for the identification of novel caspase cleavage motifs and the development of new caspase inhibitors and anti-inflammatory drugs.

Boosting of retinol activity using novel lecithin: Retinol acyltransferase inhibitors.

Lecithin:retinol acyltransferase (LRAT) is the main enzyme catalysing the esterification of retinol to retinyl esters and, hence, is of central importance for retinol homeostasis. As retinol, by its metabolite retinoic acid, stimulates fibroblasts to synthesize collagen fibres and inhibits collagen-degrading enzymes, the inhibition of LRAT presents an intriguing strategy for anti-ageing ingredients by increasing the available retinol in the skin. Here, we synthesized several derivatives mimicking natural lecithin substrates as potential LRAT inhibitors. By exploring various chemical modifications of the core scaffold consisting of a central amino acid and an N-terminal acylsulfone, we explored 10 different compounds in a biochemical assay, resulting in two compounds with IC50 values of 21.1 and 32.7 μM (compounds 1 and 2), along with a simpler arginine derivative with comparative inhibitory potency. Supported by computational methods, we investigated their structure-activity relationship, resulting in the identification of several structural features associated with high inhibition of LRAT. Ultimately, we conducted an exvivo study with human skin, demonstrating an increase of collagen III associated with a reduction of the skin ageing process. In conclusion, the reported compounds offer a promising approach to boost retinol abundance in human skin and might present a new generation of anti-ageing ingredients for cosmetic application.

Mixed-mode cationic exchange paper combined with direct infusion mass spectrometry, a sustainable approach to determine opioids in biosamples

Direct infusion mass spectrometry (DI-MS/MS) allows the rapid analysis of samples and minimizes solvent usage by avoiding chromatographic separation. However, it is prone to ion suppression due to matrix effects, especially when biosamples are processed. In this context, sample preparation appears as the perfect complement to get proper analytical results. In this article, a novel and sustainable planar sorptive phase containing mixed-mode cationic exchange (MCX) particles is presented to overcome the limitations of DI-MS/MS in bioanalysis. The new phase uses cellulose paper as a natural substrate over which the particles can be retained using polyacrylonitrile (PAN) as the chemical binder. The high porosity of the cellulose substrate simplifies the synthesis since a straightforward dip-coating technique can be applied. For this purpose, MCX particles are initially dispersed in a PAN solution prepared in N,N-Dimethylformamide. A segment of paper is later introduced into the slurry, and the evaporation of the solvent leaves a film of PAN-MCX composite over the paper, where the particles are entrapped but accessible for the analyte's isolation. The resulting MCX-paper has been evaluated for the extraction of selected opioids (codeine, methadone, morphine, naloxone, oxycodone, and tramadol) from saliva and urine samples and their determination by direct infusion tandem mass spectrometry. Working at the optimized conditions, the limits of detection were in the low μg L−1 range. The precision, expressed as relative standard deviation, was better than 12 % under intra-day and inter-day conditions. The accuracy, expressed as relative recoveries, provides values in the ranges 89–113 % and 74–121% for saliva and urine, respectively. The method was finally applied to analyze saliva and urine samples collected from patients under codeine treatment.

Quorum sensing regulating the heterogeneous transformation of antibiotic resistance genes in microplastic biofilms

Microplastics (MPs) provide a specific habitat for bacterial adhesion and the adsorption of extracellular DNA (eDNA), making them hotspots for the transformation of antibiotic resistance genes (ARGs) in the environment. However, the heterogeneous mechanism of ARG regulation by quorum sensing (QS) in MP biofilms remains unclear. This study demonstrated that QS signals, including C8-HSL, 3-oxo-C10-HSL, and C12-HSL, were detected at ng/L levels in Bacillus subtilis biofilms colonized on MPs. The addition of these three AHLs enhanced natural transformation by 1–2 orders of magnitude in both MP and natural substrate (NS) biofilms, while QS inhibitors decreased transformation frequencies by 2 orders of magnitude. Moreover, transformation frequencies in MP biofilms increased 6.0-fold under QS regulation mediated by AHLs, compared to a 2.5-fold increase in NS biofilms. QS-regulated biofilm formation and EPS secretion were found to be responsible for ARG dissemination in MP biofilms, as indicated by the positive correlation between transformation frequencies and EPS variation. Additionally, mRNA expression analysis revealed that activation and inhibition of the QS system in MP biofilms regulated the expression of natural transformation-related genes, including epsG (regulating EPS secretion) and recO (regulating eDNA transformation). These results highlight the significant influence of the QS system on ARG dissemination through heterogeneous biofilm formation and EPS secretion in MP biofilms. Understanding the effect and mechanism of QS on ARG natural transformation provides new insights into controlling ARG spread by targeting QS in host bacteria within MP biofilms.

Assay Development and Validation for Innovative Antiviral Development Targeting the N-Terminal Autoprocessing of SARS-CoV-2 Main Protease Precursors.

The SARS-CoV-2 main protease (Mpro) is initially synthesized as part of polyprotein precursors that undergo autoproteolysis to release the free mature Mpro. To investigate the autoprocessing mechanism in transfected mammalian cells, we examined several fusion precursors, with the mature SARS-CoV-2 Mpro along with the flanking amino acids (to keep the native substrate sequences) sandwiched between different tags. Our analyses revealed differential proteolysis kinetics at the N- and C-terminal cleavage sites. Particularly, N-terminal processing is differentially influenced by various upstream fusion tags (GST, sGST, CD63, and Nsp4) and amino acid variations at the N-terminal P1 position, suggesting that precursor catalysis is flexible and subject to complex regulation. Mutating Q to E at the N-terminal P1 position altered both precursor catalysis and the properties of the released Mpro. Interestingly, the wild-type precursors exhibited different enzymatic activities compared to those of the released Mpro, displaying much lower susceptibility to known inhibitors targeting the mature form. These findings suggest the precursors as alternative targets for antiviral development. Accordingly, we developed and validated a high-throughput screening (HTS)-compatible platform for functional screening of compounds targeting either the N-terminal processing of the SARS-CoV-2 Mpro precursor autoprocessing or the released mature Mpro through different mechanisms of action.

Background selection for camouflage shifts in accordance with color change in an intertidal prawn.

To maximize camouflage across visually heterogeneous habitats, animals have evolved a variety of strategies, including polyphenism, color change, and behavioral background matching. Despite the expected importance of behavioral processes for mediating camouflage, such as selection for matching substrates, behavior has received less attention than color traits themselves, and interactions between color change and behavior are largely unexplored. Here, we investigated behavioral background matching in green and red chameleon prawns (Hippolyte varians) over the course of a color change experiment. Prawns were housed on mismatching green and red seaweeds for 30 days and periodically given a choice test between the same seaweeds in y-choice trials over the experiment. We found that, as prawns change color and improve camouflage (to the perspective of a fish predator), there is a reinforcing shift in behavior. That is, as prawns shift from red to green color, or vice versa, their seaweed color preference follows this. We provide key empirical evidence that plasticity of appearance (color) is accompanied by a plastic shift in behavior (color preference) that reinforces camouflage in a color changing species on its natural substrate. Overall, our research highlights how short-term plasticity of behavior and longer-term color change act in tandem to maintain crypsis over time.

Expanding the substrate selectivity of the fimsbactin biosynthetic adenylation domain, FbsH.

Nonribosomal peptide synthetases (NRPSs) produce diverse natural products including siderophores, chelating agents that many pathogenic bacteria produce to survive in low iron conditions. Engineering NRPSs to produce diverse siderophore analogs could lead to the generation of novel antibiotics and imaging agents that take advantage of this unique iron uptake system in bacteria. The highly pathogenic and antibiotic-resistant bacteria Acinetobacter baumannii produces fimsbactin, an unusual branched siderophore with iron-binding catechol groups bound to a serine or threonine side chain. To explore the substrate promiscuity of the assembly line enzymes, we report a structure-guided investigation of the stand-alone aryl adenylation enzyme FbsH. We report on structures bound to its native substrate 2,3-dihydroxybenzoic acid (DHB) as well as an inhibitor that mimics the adenylate intermediate. We produced enzyme variants with an expanded binding pocket that are more tolerant for analogs containing a DHB C4 modification. Wild-type and mutant enzymes were then used in an in vitro reconstitution analysis to assess the production of analogs of the final product as well as several early-stage intermediates. This analysis shows that some altered substrates progress down the fimsbactin assembly line to the downstream domains. However, analogs from alternate building blocks are produced at lower levels, indicating that selectivity exists in the downstream catalytic domains. These findings expand the substrate scope of producing condensation products between serine and aryl acids and identify the bottlenecks for chemoenzymatic production of fimsbactin analogs.

Factors influencing the epibiota of granite blocks used in coastal defense structures: An observational experiment

Anthropogenic pressures exerted on coastal areas, combined with climate change, and accelerated sea level rise, result in the loss of natural habitats. At the same time, the number of coastal protection and defense structures have increased considerably and may become the dominant habitat in many intertidal regions. In this study we characterize the epibiota of artificial substrates (granite blocks) from coastal defense structures used to contain the effects of rising sea levels, and the same type of artificial substrate found in contact with coastal reefs located on Serrambi beach, northeast of Brazil. We mapped, measured, and photographed 45 artificial granite blocks and their surroundings in the field. A gradient was found for the number of taxa on the blocks, from high coverage by macroalgae to extensive non-colonized areas. However, it was observed that the closer to the natural reef the greater the taxonomic richness. Our results also indicate that the factor that best explains the colonization of blocks is the coverage of their surroundings, thus, few organisms were found on blocks whose surrounding unconsolidated sediment area was very extensive. In our wind and sun exposure simulation, granite rock demonstrated a more accelerated and greater level of heating (up to 3 °C) compared to fragments of natural substrate, although the variation in the slightly lighter colored and smaller granite block resembles the variation in the reef fragment. We suggest that the use of lighter-colored granite blocks at mid-coastal levels can increase the colonization rate of these substrates.