Engineering Applications Research Articles

Synergistic effects of Monel 400 filler wire in gas metal arc welding of CoCrFeMnNi high entropy alloy

Weldability plays a crucial role in the journey of high entropy alloys towards their engineering applications. In this study, gas metal arc welding was performed to join an as-rolled CoCrFeMnNi high entropy alloy using Monel 400 as the filler wire. The present research findings demonstrate a favorable metallurgical chemical reaction between the Monel 400 filler and the CoCrFeMnNi high entropy alloy, resulting in compositional mixing within the fusion zone that promotes a solid-solution strengthening effect, counteracting the typical low hardness associated to the fusion zone of these alloys. The weld thermal cycle induced multiple microstructure changes across the joint, including variations in the grain size, existing phases and local texture. The grain size was seen to increase from the base material toward the fusion zone. An FCC matrix and finely sparse Cr-Mn-based oxides existed in both base material and heat affected zone, while in the fusion zone new FCC phases and carbides were formed upon the mixing of the Monel 400 filler. The role of the filler material on the fusion zone microstructure evolution was rationalized using thermodynamic calculations. Texture shifted from a γ-fiber (in the base material) to a strong cubic texture in the fusion zone. Digital image correlation during tensile testing to fracture coupled with microhardness mapping revealed that, stemming from the process-induced microstructure changes, the micro and macromechanical response differed significantly from the original base material. This study successfully established a correlation between the impact of the process on the developed microstructural features and the resultant mechanical behavior, effectively assessing the processing-microstructure-properties relationships towards an improved understanding of the physical metallurgy associated to these advanced engineering alloys. In conclusion, this work provides an important theoretical framework and practical guidance for optimizing the engineering applications of high entropy alloys.

Fault diagnosis method of PEMFC system based on ensemble learning

Due to the coupling of multiple physical fields inside proton exchange membrane fuel cell (PEMFC) and the complex and changeable external application scenarios, the faults of PEMFC system greatly affects its engineering application. The paper proposes an ensemble learning-based fault diagnosis method for PEMFC systems, utilizing five distinct algorithms to develop an integrated ensemble classifier for fault detection. The original sample dataset collected from fuel cells undergoes a process of deep feature extraction to obtain a composite feature sample dataset, which is then inputted into an ensemble classifier to establish a fault diagnosis model. The experimental data of four kinds of faults of 80 W fuel cell test bench, EC fuel cell system and 50 W water-cooled fuel cell test bench are analyzed by using this classification model. The results show that the accuracy and generalization of the ensemble learning algorithm for different fault diagnosis of PEMFC system in different application environments are verified.

Two-scale finite element simulations for UT inspection of micro-structured materials in CIVA

Ultrasonic testing (UT) of highly heterogeneous structures is an important issue for many industrial applications in civil engineering for concrete inspection as well as in nuclear and oil & gas, where it applies to cast steels and weld joints. In these materials, performance is limited by the phenomenon of multiple wave scattering at the microstructure scale. In this context, the use of effective simulation tools is particularly useful both for improving interpretation of UT and for optimizing or demonstrating the performance of inspections. However, the scales of heterogeneity to be taken into account are often smaller than fractions of a tenth of a wavelength. In 3D simulations, it is therefore difficult, if not impossible, to combine computational performances with the refinement of the modelled interactions. To meet this challenge, CEA is developing an original approach based on two applications of the finite element (FE) method implemented in CIVA. The first application uses micro-scale heterogeneous material data on small representative elemental volumes (REVs) to feed a second code dedicated to macro-scale with effective medium characteristic. This second code relies on a high-order finite element model that is very efficient at the scale of the inspected industrial component. In this communication we present the main features of these FE tools and illustrate their performances on different application cases for welded parts and concrete

Knowledge graph and development hotspots of biochar as an emerging aquatic antibiotic remediator: A scientometric exploration based on VOSviewer and CiteSpace

As an emerging material in the field of environmental remediation, biochar produced by carbonisation of organic solid waste has been widely used in the remediation of antibiotic wastewater due to its environmental friendliness and excellent adsorption properties. This study analyses the current literature in the field in a comprehensive and scientific manner using CiteSpace and VOSviewer technologies. Between 2011 and 2023, a total of 1162 papers were published in this domain, spanning three distinct stages: applied methods, mechanism investigation, and enhanced improvement. The results of keyword clustering indicate that the remediation of antibiotics complexed with multiple pollutants by biochar is the main research topic, followed by the remediation of antibiotics by biochar in combination with other technologies. Furthermore, drawing from current research hotspots in antibiotic remediation using biochar, this study identified the pivotal mechanisms involved: (1) The primary mechanisms by which raw biochar remediates antibiotics include π-π electron donor-acceptor interactions, hydrophobic interactions, electrostatic interactions, hydrogen-bonding, and pore filling. (2) Steam activation, acid/base, metal salt/metal oxide, and clay mineral modification can improve the physical/chemical properties of biochar, enhancing its adsorptive removal of antibiotics. (3) Biochar activated persulfate and degraded antibiotics via free radical pathways (SO4−•, •OH and O2−•) as well as non-free radical pathways (1O2 and electron transfer). In addition, the challenge and prospect of biochar engineering applications for antibiotic remediation lies in improving the main mechanism of antibiotic remediation by biochar. The prospective utilization of biochar in enhancing the remediation of antibiotic-related pollutants holds tremendous value for the future.

Stimuli-Responsive Biomaterials for Tissue Engineering Applications.

The use of 'smart materials,' or 'stimulus-responsive' materials, has proven useful in a variety of fields, including tissue engineering and medication delivery. Many factors, including temperature, pH, redox-state, light, and magnetic fields, are being studied for their potential to affect a material's properties, interactions, structure, and/or dimensions. New tissue engineering and drug delivery methods are made possible by the ability of living systems to respond to both external stimuli and their own internal signals (for example, materials composed of stimuli-responsive polymers that self-assemble or undergo phase transitions or morphology transformation. The researcher examines the potential of smart materials as controlled drug release vehicles in tissue engineering, aiming to enable the localized regeneration of injured tissue by delivering precisely dosed drugs at precisely timed intervals.

Anti-seepage performance and underlying mechanism of carboxymethyl cellulose sodium-modified bentonite in hydraulic engineering applications

Bentonite is widely utilized in hydraulic engineering projects. However, its performance may deteriorate under extreme environmental conditions. Hence, we aim to develop a sodium carboxymethyl cellulose (CMC)-modified bentonite and investigate its anti-seepage performance. The results showed that the 10% CMC-modified bentonite exhibited the best performance. Compared with unmodified bentonite, the expansion index of 10% CMC-modified bentonite significantly increased to 40 mL/2 g, indicating a high water absorption capacity and expansion property. Moreover, it showed a significant decrease in permeability coefficient (3.72×10−11 m/s) and an increase in porosity (1.90), indicating its excellent anti-seepage effect. Furthermore, under unconventional environmental conditions and long-term performance tests, the 10% CMC-modified bentonite shows improved stability compared to the unmodified bentonite. Therefore, it can be concluded that this study provides support for the utilization of CMC-modified bentonite in anti-seepage engineering and hydraulic engineering applications.

Design of Variable Valve Timing and Electronic Control for Honda R18A

Abstract: This paper explores the Variable Valve Timing and Lift Electronic Control (VTEC) system developed by Honda, focusing on its mechanisms, design methodology, and impact on engine performance. VTEC optimizes engine efficiency and power by switching between cam profiles suited for low-RPM stability and high-RPM power output, controlled by the engine's computer based on operational conditions. We examine the intricate workings of the VTEC system, detailing its hydraulic actuation and the resulting enhancements in torque and horsepower. Our study centers on the Honda R18A engine, targeting increased maximum RPM and power output. Using theoretical analysis validated through Ricardo Wave software and MATLAB simulations, we determine optimal valve lift and timing to achieve the desired performance. Additionally, the research addresses the geometric parameters influencing airflow through the valves, providing a comprehensive understanding of the VTEC system's contribution to engine efficiency and driving experience. This investigation not only underscores the technological advancements in variable valve timing but also presents practical insights for automotive engineering applications.

Sustainable End-of-Life Tyre Management: A Comprehensive Analysis of Environmental Impacts and Crumb Rubber Integration in Composite Concretes

The research explores pollution prevention strategies associated with end-of-life tyres (ELTs) through sustainable practices, focusing on repurposing ELTs into crumb rubber (CR) for use in composite concretes for civil engineering applications. Three disposal approaches are considered: recycling into crumb rubber for use in cement composites to enhance acoustic properties, incineration for cement production, and landfill disposal. The study aims to assess the environmental impact of each method, particularly in terms of carbon dioxide (CO2) emissions during the recycling phase, utilizing the OpenLCA program for a comprehensive life cycle analysis. The primary objective is to provide insights into the sustainability of incorporating recycled rubber in concrete mixes as fine aggregate to improve concrete acoustic and shock absorbance properties. The research also gathers data on CO2 emissions from ELT incineration for cement production and the environmental implications of ELT’s landfill disposal. By comparing these three strategies, the study offers a holistic perspective on the environmental ramifications of ELT management. Notably, a recent study highlights the energy recovery and CO2 emissions from ELT incineration, demonstrating the potential benefits of recycling. The research identifies a gap in existing studies, emphasizing the need to consider the entire life cycle, including the transportation and use stages. Notably, the use stage significantly contributes to environmental impacts, with carbon emissions ranging from 550 to 840 kg CO2 eq., per car tyre. Additionally, a proposed inventory analysis method evaluates greenhouse gas emissions from Portland cement concrete pavement construction, revealing that raw material production accounts for most emissions. The findings of this research are providing valuable insights for policymakers, industry professionals, and environmentalists aiming for sustainable practices in the construction sector. By allocating a fraction of tyre production CO2 emissions to obtain raw material for concrete composites, the study suggests a potential avenue for reducing environmental impact. Further calculations using the OpenLCA program may refine these recommendations.

Application of microbially induced calcium carbonate precipitation (MICP) process in concrete self-healing and environmental restoration to facilitate carbon neutrality: a critical review.

Soil contamination, land desertification and concrete cracking can have significant adverse impacts on sustainable human economic and societal development. Cost-effective and environmentally friendly approaches are recommended to resolve these issues. Microbially induced carbonate precipitation (MICP) is an innovative, attractive and cost-effective in situ biotechnology with high potential for remediation of polluted or desertified soils/lands and cracked concrete and has attracted widespread attention in recent years. Accordingly, the principles of MICP technology and its applications in the remediation of heavy metal-contaminated and desertified soils and self-healing of concrete were reviewed in this study. The production of carbonate mineral precipitates during the MICP process can effectively reduce the mobility of heavy metals in soils, improve the cohesion of dispersed sands and realize self-healing of cracks in concrete. Moreover, CO2 can be fixed during MICP, which can facilitate carbon neutrality and contribute to global warming mitigation. Overall, MICP technology exhibits great promise in environmental restoration and construction engineering applications, despite some challenges remaining in its large-scale implementation, such as the substantial impacts of fluctuating environmental factors on microbial activity and MICP efficacy. Several methods, such as the use of natural materials or wastes as nutrient and calcium sources and isolation of bacterial strains with strong resistance to harsh environmental conditions, are employed to improve the remediation performance of MICP. However, more studies on the efficiency enhancement, mechanism exploration and field-scale applications of MICP are needed.

Unsupervised learning of quantum many-body scars using intrinsic dimension

Quantum many-body scarred systems contain both thermal and non-thermal scar eigenstates in their spectra. When these systems are quenched from special initial states which share high overlap with scar eigenstates, the system undergoes dynamics with atypically slow relaxation and periodic revival. This scarring phenomenon poses a potential avenue for circumventing decoherence in various quantum engineering applications. Given access to an unknown scar system, current approaches for identification of special states leading to non-thermal dynamics rely on costly measures such as entanglement entropy. In this work, we show how two dimensionality reduction techniques, multidimensional scaling and intrinsic dimension estimation, can be used to learn structural properties of dynamics in the PXP model and distinguish between thermal and scar initial states. The latter method is shown to be robust against limited sample sizes and experimental measurement errors.

Loom: A Modular Open-Source Approach to Rapidly Produce Sensor, Actuator, Datalogger Systems

In the face of rising population, erratic climate, resource depletion, and increased exposure to natural hazards, environmental monitoring is increasingly important. Satellite data form most of our observations of Earth. On-the-ground observations based on in situ sensor systems are crucial for these remote measurements to be dependable. Providing open-source options to rapidly prototype environmental datalogging systems allows quick advancement of research and monitoring programs. This paper introduces Loom, a development environment for low-power Arduino-programmable microcontrollers. Loom accommodates a range of integrated components including sensors, various datalogging formats, internet connectivity (including Wi-Fi and 4G Long Term Evolution (LTE)), radio telemetry, timing mechanisms, debugging information, and power conservation functions. Additionally, Loom includes unique applications for science, technology, engineering, and mathematics (STEM) education. By establishing modular, reconfigurable, and extensible functionality across components, Loom reduces development time for prototyping new systems. Bug fixes and optimizations achieved in one project benefit all projects that use Loom, enhancing efficiency. Although not a one-size-fits-all solution, this approach has empowered a small group of developers to support larger multidisciplinary teams designing diverse environmental sensing applications for water, soil, atmosphere, agriculture, environmental hazards, scientific monitoring, and education. This paper not only outlines the system design but also discusses alternative approaches explored and key decision points in Loom’s development.

Toward Bioactive Hydrogels: A Tunable Approach via Nucleic Acid-Collagen Complexation

Abstract Purpose Nucleic acid-collagen complexes (NACCs) are unique biomaterials formed by binding short, monodisperse single-stranded DNA (ssDNA) with type I collagen. These complexes spontaneously generate microfibers and nanoparticles of varying sizes, offering a versatile platform with potential applications in tissue engineering and regenerative medicine. However, the detailed mechanisms behind the nucleic acid-driven assembly of collagen fibers still need to be established. We aim to understand the relationship between microscopic structure and bulk material properties and demonstrate that NACCs can be engineered as mechanically tunable systems. Methods We present a study to test NACCs with varying molar ratios of collagen to random ssDNA oligonucleotides. Our methods encompass the assessment of molecular interactions through infrared spectroscopy and the characterization of gelation and rheological behavior. We also include phase contrast, confocal reflectance, and transmission electron microscopy to provide complementary information on the 3D structural organization of the hydrogels. Results We report that adding DNA oligonucleotides within collagen robustly reinforces and rearranges the hydrogel network and accelerates gelation by triggering rapid fiber formation and spontaneous self-assembly. The elasticity of NACC hydrogels can be tailored according to the collagen-to-DNA molar ratio, ssDNA length, and collagen species. Conclusion Our findings hold significant implications for the design of mechanically tunable DNA-based hydrogel systems. The ability to manipulate hydrogel stiffness by tailoring DNA content and collagen concentration offers new avenues for fine tuning material properties, enhancing the versatility of bioactive hydrogels in diverse biomedical applications. Lay Summary This work is an example of forming fibers and gels with tunable elasticity that stems from the complexation of short-length nucleic acids (on the order of size of aptamers) and collagen, which can be potentially extended to a variety of functionalized hydrogel designs and tailored biomedical applications. Incorporating DNA induces mechanical changes in NACCs. Graphical Abstract

Riboflavin modified carbon cloth enhances anaerobic digestion treating food waste in a pilot-scale system

Previous laboratory-scale studies have consistently shown that carbon-based conductive materials can notably improve the anaerobic digestion of food waste, typically employing reactors with regular capacity of 1–20 L. Furthermore, incorporating riboflavin-loaded conductive materials can further address the imbalance between fermentation and methanogenesis in anaerobic systems. However, there have been few reports on pilot-scale investigation. In this study, a 10 m2 of riboflavin modified carbon cloth was incorporated into a pilot-scale (2 m3) food waste anaerobic reactor to improve its treatment efficiency. The study found that the addition of riboflavin-loaded carbon cloth can increase the maximum organic loading rate (OLR) by 40% of the pilot-scale reactor, compared to the system using carbon cloth without riboflavin loading, while ensuring efficient operation of the reaction system, effectively alleviating system acidification, sustaining methanogen activity, and increasing daily methane production by 25%. Analysis of the microbial community structure revealed that riboflavin-loaded carbon cloth enriched the methanogenic archaea in the genera of Methanothrix and Methanobacterium, which are capable of extracellular direct interspecies electron transfer (DIET). And metabolic pathway analysis identified the methane production pathway, highly enriched on the reduction of acetic acid and CO2 at riboflavin-loaded carbon cloth sample. The expression levels of genes related to methane production via DIET pathway were also significantly upregulated. These results can provide important guidance for the practical application of food waste anaerobic digestion engineering.

Crosstalk among miR-29, α-SMA, and TGFβ1/β3 in melatonin-induced exosome (Mel-prExo) treated human limbal mesenchymal stem cells (hLMSCs): An insight into scarless healing of the cornea.

Inflammatory mediators that infiltrate the corneal stroma after corneal infections, trauma or refractive surgery can trigger the transformation of corneal keratocytes into myofibroblasts, resulting in highly irregular collagen deposition and subsequently corneal scarring. Mesenchymal stem cells (MSCs) can be used as therapeutic agents to regenerate corneal and conjunctival tissue damage, regulate inflammation, and reduce the development of limbal stem cell failure. The use of MSC-derived exosomes as a cell-free therapeutic vector is a novel therapeutic approach. This study aimed to assess the effect of exosomes obtained from melatonin (Mel)-treated human limbal mesenchymal stem cells (hLMSCs) on naïve hLMSCs and to determine their influence on the antifibrotic and pro-regenerative pathways involved in corneal scarring. hLMSCs were treated with varying concentrations of Mel, followed by isolation and characterization of the procured exosomes (Mel-prExos). These exosomes were added to the cell culture media of naïve hLMSCs to examine their antifibrotic and pro-regenerative effects. The expression of miR-155, miR-29, TGFβ1, TGFβ3, PPARγ, and α-SMA miRNAs and genes were compared between Mel-treated hLMSCs and Mel-prExo-treated hLMSCs by using real-time PCR. We found that at 1 μM Mel and in the presence of Mel-prExos, TGFβ1 was expressed 0.001-fold, while TGFβ3 was expressed 0.6-fold. miR-29 expression was increased 38-fold in the control-Exo group compared to that in the control group. Changes in TGFβ1/β3 and α-SMA expression are associated with miR-29 and miR-155. This approach could prove beneficial for ocular surface tissue engineering applications.

Enhancing Load-Bearing Capacity of Calcareous Sands through Gel Stabilization: A Mechanical and Material Characterization Study

Calcareous sands often display wide ring grain configurations, high intragranular porosity, a complex structure, and low grain hardness. These attributes typically do not meet the strength criteria necessary to sustain overlying infrastructure in civil engineering applications. This study investigates gel stabilization techniques, blending gel material with calcareous sand at concentrations ranging from 5% to 22%, followed by curing periods of 3 to 28 days to evaluate the load-bearing capacity. Subsequently, an unconfined compressive test is performed to determine the gel material content in stabilized specimens and investigate the influence of gel material types. The gel material-to-sand ratios employed are set at 5%, 10%, and 16% for Portland cement and 13%, 16%, and 22% for gypsum. After that, a triaxial consolidated undrained test is conducted to assess mechanical behavior, pore water pressure, and mechanical properties. The findings reveal increased dilation, stress–strain hardening, and softening post-yield, regardless of gel material type. Principal stress ratios, secant modulus, and cohesion show a positive correlation with maintenance duration and binder content, with implications for improved load-bearing capacity. The study also elucidates the qualitative relationship between secant modulus E50 and confining pressure.

Neodymium (III)-containing poly(lactide-co-glycolide)-coated robocast bioactive glass scaffolds for photothermal therapy and bone regeneration

In this study, trivalent neodymium-doped silicate-based 13–93 bioactive glass scaffolds were prepared by the robocasting method using sol-gel-derived bioactive glass powders for tissue engineering applications. Sintered scaffolds were coated by borate-based 13-93B3 bioactive glass-containing polylactide-co-glycolide solution. The produced composite scaffolds’ mechanical, morphological, and structural characteristics were thoroughly examined, as their in vitro bioactivity in cell culture media and simulated body fluid. Furthermore, the scaffolds’ amoxicillin adsorption and release behavior was examined over time. The outcomes demonstrated that it was feasible to effectively create periodic, mesh-like-patterned robocast glass scaffolds utilizing Nd3+-doped sol-gel-derived bioactive glass powders. The scaffolds’ compressive strengths ranged from 10.02 MPa to 18.6 MPa, with the PLGA-coated scaffolds exhibiting the highest strength values. All of the scaffolds that were submerged in simulated body fluid for 28 days showed hydroxyapatite formation. The presence of borate glass on the surface of the silicate-based glass scaffolds improved the hydroxyapatite formation ability. The quantity of drug adsorption for all types of scaffolds was measured to be between 4 and 9% whereas the cumulative drug release was in the range of 58 to 96%. Borate glass particle-containing PLGA coating enhanced the drug delivery behavior.

Design of overhead ground wire repair robot based on unmanned aerial vehicles

AbstractAiming at the problem that it is impossible to accurately transport the aerial ground wire repair robot to the repair point and efficiently repair the ground wire broken strands and scattered strands, this paper designs an aerial ground wire repair robot based on Unmanned Aerial Vehicle (UAV). Firstly, the mechanical structure of the overhead ground wire repair robot is selected and designed, and the information transmission and the cooperative operation of each module are realized. A landing mark based on ArUco code is designed, which uses C++ for visual navigation, and then uses the double closed‐loop Proportional Integral Derivative (PID) control method to control the position and attitude of the UAV, so as to realize the tracking of the robot position by the UAV. At the same time, the mechanical structure and the control method of the UAV hanging device are designed to cooperate with the overhead ground wire repair robot to ensure the accurate completion of the repair operation. Finally, a UAV‐based aerial ground wire repair robot prototype was developed and successfully applied to the repair operation of the broken strand of the aerial ground wire of the 66 to 67 tower of the 500 kV Jiangzi line in Ya'an. It is verified that the UAV‐based aerial ground wire repair robot studied in this paper has a good engineering application value.

Polyetheretherketone bioactivity induced by farringtonite

Polyetheretherketone (PEEK) is considered as an excellent biomaterial for bone grafting and connective tissue replacement. The clinical potential is, however, limited by its bioinertness, poor osteoconduction, and weak antibacterial activity. These disadvantages can be overcome by introducing suitable additives to produce mineral-polymer composites or coatings. In this work, a PEEK-based bioactive composite has been obtained by blending the polymer with magnesium phosphate (Mg3(PO4)2) particles in amounts ranging from 1 to 10 wt.% using the hot press technique. The obtained composite exhibited improved mechanical and physical properties, above the lower limits set for bone engineering applications. The tested grafts were found to not induce cytotoxicity. The presence of magnesium phosphate induced the mineralisation process with no adverse effects on the expression of the marker crucial for osteoblastic differentiation. The most promising results were observed in the grafts containing 1 wt.% of magnesium phosphate embedded within the PEEK matrix. The improved bioactivity of grafts, together with suitable physical–chemical and mechanical properties, indicate this composite as a promising orthopaedic implant material.

Ultrahigh strength-ductility synergy via heterogeneous grain structure and multi-scale L12-γ′ precipitates in a cobalt-based superalloy GH159

Achieving excellent strength-ductility synergy through convenient techniques is highly desirable for modern engineering applications. In this study, we constructed a heterogeneous grain structure consisting of large residual deformed grains and fine recrystallized grains, along with multi-scale L12-γ′ precipitates, in a commercial cobalt-based superalloy GH159. This was achieved via short-term annealing followed by subsequent aging. The mechanical properties were significantly enhanced, demonstrating an exceptional ultimate tensile strength of 1611 MPa, and outstanding uniform and total elongations of 23.5 % and 34.7 %, respectively. We investigated the evolution of the dual heterogeneous structures and the corresponding strengthening mechanisms during deformation. Hetero-deformation induced (HDI) strengthening, L12-γ′ precipitation strengthening, along with nanotwins, L-C locks, and 9R phases contribute significantly to the excellent strength-ductility synergy in GH159. These findings offer a strategy for developing GH159 as a structural material for various engineering applications, providing accessible and cost-effective processing methods that pave the way for large-scale industrial production. Furthermore, this study provides design insights for enhancing the mechanical properties of cobalt-based precipitation-strengthened superalloys.

Investigation of the Odilorhabdin Biosynthetic Gene Cluster Using NRPS Engineering.

The recently identified natural product NOSO-95A from entomopathogenic Xenorhabdus bacteria, derived from a biosynthetic gene cluster (BGC) encoding a non-ribosomal peptide synthetase (NRPS), was the first member of the odilorhabdin class of antibiotics. This class exhibits broad-spectrum antibiotic activity and inspired the development of the synthetic derivative NOSO-502, which holds potential as a new clinical drug by breaking antibiotic resistance. While the mode of action of odilorhabdins was broadly investigated, their biosynthesis pathway remained poorly understood. Here we describe the heterologous production of NOSO-95A in Escherichia coli after refactoring the complete BGC. Since the production titer was low, NRPS engineering was applied to uncover the underlying biosynthetic principles. For this, modules of the odilorhabdin NRPS fused to other synthetases were co-expressed with candidate hydroxylases encoded in the BGC allowing the characterization of the biosynthesis of three unusual amino acids and leading to the identification of a prodrug-activation mechanism by deacylation. Our work demonstrates the application of NRPS engineering as a blueprint to mechanistically elucidate large or toxic NRPS and provides the basis to generate novel odilorhabdin analogues with improved properties in the future.