Engineering Methods Research Articles

A Robust α-l-Fucosidase from Prevotella nigrescens for Glycoengineering Therapeutic Antibodies.

Eliminating the core fucose from the N-glycans of the Fc antibody segment by pathway engineering or enzymatic methods has been shown to enhance the potency of therapeutic antibodies, especially in the context of antibody-dependent cytotoxicity (ADCC). However, there is a significant challenge due to the limited defucosylation efficiency of commercially available α-l-fucosidases. In this study, we report a unique α-l-fucosidase (PnfucA) from the bacterium Prevotella nigrescens that has a low sequence identity compared with all other known α-l-fucosidases and is highly reactive toward a core disaccharide substrate with fucose α(1,3)-, α (1,4)-and α(1,6)-linked to GlcNAc, and is less reactive toward the Fuc-α(1,2)-Gal on the terminal trisaccharide of the oligosaccharide Globo H (Bb3). The kinetic properties of the enzyme, such as its Km and kcat, were determined and the optimized expression of PnfucA gave a yield exceeding 30 mg/L. The recombinant enzyme retained its full activity even after being incubated for 6 h at 37 °C. Moreover, it retained 92 and 87% of its activity after freezing and freeze-drying treatments, respectively, for over 28 days. In a representative glycoengineering of adalimumab (Humira), PnfucA showed remarkable hydrolytic efficiency in cleaving the α(1,6)-linked core fucose from FucGlcNAc on the antibody with a quantitative yield. This enabled the seamless incorporation of biantennary sialylglycans by Endo-S2 D184 M in a one-pot fashion to yield adalimumab in a homogeneous afucosylated glycoform with an improved binding affinity toward Fcγ receptor IIIa.

An Overview on the Physiopathology of the Blood-Brain Barrier and the Lipid-Based Nanocarriers for Central Nervous System Delivery.

The state of well-being and health of our body is regulated by the fine osmotic and biochemical balance established between the cells of the different tissues, organs, and systems. Specific districts of the human body are defined, kept in the correct state of functioning, and, therefore, protected from exogenous or endogenous insults of both mechanical, physical, and biological nature by the presence of different barrier systems. In addition to the placental barrier, which even acts as a linker between two different organisms, the mother and the fetus, all human body barriers, including the blood-brain barrier (BBB), blood-retinal barrier, blood-nerve barrier, blood-lymph barrier, and blood-cerebrospinal fluid barrier, operate to maintain the physiological homeostasis within tissues and organs. From a pharmaceutical point of view, the most challenging is undoubtedly the BBB, since its presence notably complicates the treatment of brain disorders. BBB action can impair the delivery of chemical drugs and biopharmaceuticals into the brain, reducing their therapeutic efficacy and/or increasing their unwanted bioaccumulation in the surrounding healthy tissues. Recent nanotechnological innovation provides advanced biomaterials and ad hoc customized engineering and functionalization methods able to assist in brain-targeted drug delivery. In this context, lipid nanocarriers, including both synthetic (liposomes, solid lipid nanoparticles, nanoemulsions, nanostructured lipid carriers, niosomes, proniosomes, and cubosomes) and cell-derived ones (extracellular vesicles and cell membrane-derived nanocarriers), are considered one of the most successful brain delivery systems due to their reasonable biocompatibility and ability to cross the BBB. This review aims to provide a complete and up-to-date point of view on the efficacy of the most varied lipid carriers, whether FDA-approved, involved in clinical trials, or used in in vitro or in vivo studies, for the treatment of inflammatory, cancerous, or infectious brain diseases.

A software for calculating coal mine gas emission quantity based on the different-source forecast method

The ability to predict gas emissions accurately is pivotal in managing gas control and ensuring safe mining operations. Existing internationally acknowledged gas control and prediction software does not cater to the specific conditions in Chinese coal mines. Hence, this paper introduces an object-oriented programming method to design a software tool for calculating the total gas emission quantity using the MATLAB application program designer runtime environment. The software incorporates an algorithm, data structure, framework, and module functions, all of which enable seamless integration and visualization of gas emission calculation software. This software tool mitigates the inefficiencies and inaccuracies associated with manual, different-source forecast methods. Based on the field data of the Hulonggou Coal Mine in Shanxi province, this technical software was used to predict the gas emission of the mine. The research results show that the predicted value of the technical software is close to the actual measured value. The differing estimates of the working face and coal mine output primarily account for the deviation between the tool's predicted gas emission value and the field-measured value. The underlying design logic of this technical software determines that it has good adaptability to mines with clear mining technology parameters and gas geological parameters. This study provides a valuable method for researchers and engineers seeking to improve gas emission calculation efficiency.

Valence Engineering Boosts Kinetics and Storage Capacity of Layered Double Hydroxides for Aqueous Magnesium‐Ion Batteries

AbstractThe kinetics and storage‐capacity of NiCoMg‐ternary layered double hydroxide (NiCoMg‐LDH) are successfully boosted by valence engineering. As the cathode for aqueous magnesium‐ion batteries (AMIBs), the assembled NiCoMg‐LDH//active carbon (AC) delivers a high specific discharge capacity (121.0 mAh·g−1 at 0.2 A·g−1), long‐term cycling stability (85% capacity retention after 2000 cycles at 1.0 A·g−1) and an excellent performance at −30 °C. Moreover, NiCoMg‐LDH//perylenediimide (PTCDI) is assembled, achieving a high specific discharge capacity and long‐term cycling stability. X‐ray absorption spectra (XAS)/X‐ray photoelectron spectroscopy (XPS) analyses and Density functional theory (DFT) calculations disclose that the electrons are redistributed due to the 3d orbital overlap of Co/Ni atoms in NiCoMg‐LDH, which obviously reduces the valence states of Co/Ni atoms, enhances Mg─O bond strength and degree of hybridization of Co/Ni 3d and O 2p orbitals. Hence, the electronic conductivity is significantly enhanced and the electrostatic repulsion between Mg2+ and host layers is greatly reduced, giving rise to the improved diffusion kinetics and storage‐capacity of Mg2+. Furthermore, in situ Raman/X‐ray diffraction (XRD) and ex situ XPS reveal corresponding energy‐storage mechanism. This paper not only demonstrates the feasibility of LDHs as cathode for AMIBs, but also offers a new modification method of valence engineering for high‐performance electrode materials.

Roadmap Analysis of Artificial Intelligence Engineering Method

Roadmap Analysis of Artificial Intelligence Engineering Method

CLIMATE AND ENVIRONMENTAL BACKGROUNDS OF FUEL UTILIZATION, INFLUENCING UPON ALTERATION THE EUROPEAN AND UKRAINIAN TRENDS OF GAS SUPPLY. PART 2. PROVIDING THE EFFICIENCY OF FUEL USE IN CONDITIONS OF DECARBONIZATION OF ENVIRONMENT

The paper is dedicated to power and environmental aspects of option the most appropriate types of fuel for supplying the economics of European countries and Ukraine. Accordingly notions about influence of C-containing fuels on greenhouse effect and on warming of environment, the existing tendency of substitution the fossil fuels — generally — and the natural gas — particularly — for methane-hydrogen mixtures is discussed as a controversial point. The problem of advancement the national fuel complexes is under consideration and has been studied with approach based upon the example of Ukrainian pre-war state of economics. An attention has been paid up on reduction of production and consumption of all types of the fossil fuels in the last decades, mainly due lowering the industrial using constituent. An enhancement of fuel using efficiency due improvement of furnace equipment basis provides an successive solution the fuel problems. The thermodynamic background for evaluation the efficiency of fuel using with account of the furnace’s non-ideality has been developed. The problem of “wet combustion” takes of special significance by the reason of weather and climate conditions’ account, on the one hand, and an opportunity to control the efficiency of fuel using, on the other hand, with new combined technique. The contemporary approach consists in combination the direct method of decrease the CO2 emission due exclusion of using the C-containing fuels (firstly — the fossil fuels) and with enhancement the efficiency of using the fossil fuels, for example due erection the recuperative heat recovery facilities with preliminary preheating of combustion air, in some advanced cases by combination with an air, humidification. The perspective ways of provision the decarbonization of environment consists in substitution the fossil fuels (mainly the natural gas NG) for hydrogen or H2-containing gas mixtures. The thermodynamic analysis of relevant technique has been carried out to estimate the opportunities of control the efficiency of hydrogen admixture to NG. Taking into account the variety of technologies of hydrogen production, the engineering method with giving of conditional coloration of hydrogen as a fuel has been considered. The “green” H2 makes the most valuable type of fuel (compared with “blue”, “turquoise”, “yellow”, “rose” colored and especially to the “grey” H2) as a substituting fuel from the standpoint of H2 fitness to process and plant. Particularly for high-temperature melting furnaces a provision of maximum emissivity of flame requires an using of carbon containing fuels to organize thermal pyrolysis of gas and soot formation within the combustion zone. Ref. 44, Fig. 5, Tab. 3.

SYNTHESIS OF RATIONAL CONSTRUCTIVE SOLUTION OF STEEL ROOF TRUSSES

The article examines the issue of the occurrence and influence of bending moments on the bearing capacity of a combined steel truss made of rectangular bent-welded profiles. A comparative analysis of technical and economic indicators in terms of material intensity, labor intensity, and cost of traditional-typical and lightweight combined steel trusses was carried out. The paper examines steel trusses of long-span buildings with a span of 30 m. The force graphs (bending moments and axial forces) for various calculation schemes of the combined steel truss are shown. An analysis of the forces in the combined truss for different ways of connecting the lattice to the chords was carried out. The plot of normal stresses along the middle line of the stiffening girder of the combined truss for various calculation schemes is presented. The load-bearing capacity of compressed rods with different connection methods was calculated. According to the research results, an engineering method of taking into account the influence of bending moments is proposed.

Підхід до визначення напружено-деформованого стану стрижнів з композиційних матеріалів з урахуванням внутрішнього самоврівноваженого напруженого стану

The subject of this research is methods of stress-strain state analysis of composite structures. The goal is to develop a mechanism to explain the phenomenon of warping of thin-walled composite sections with a non-uniform cross-section under thermal loading and to synthesize a model to determine the forces in the section elements. The objectives of this study are to predict the deformed states of composite sections at the manufacturing stage and to determine the temperature stresses that occur at the manufacturing stage, as well as after the assembly of the structure. The methods used are related to the mechanics of materials and structures. The following results were obtained. The possible deformation forms of composite plates with an asymmetric structure are analyzed and the rationale for the appearance of such deformations is given. A model of the force interaction of elements in a folded section is proposed. The proposed model can be extended to most sections of load-carrying aircraft structures. Based on the analysis of the mechanics of thin-walled folded sections, the mechanism of the occurrence of complex bending-torsional deformation of long-dimensional sections made of composites with a non-uniform cross-section under the action of an internal self-equilibrium stress state caused by a change in temperature or shrinkage of the binder or both is explained. The analysis of this mechanism makes it possible to design composite sections with minimal warping and to propose engineering methods to compensate for unwanted spatial movements in the case of arbitrary loading of section elements. Conclusions. The scientific novelty of the obtained results can be summarized as follows. A mechanism for the deformation of rods made of dissimilar composite materials when the temperature changes (after extraction from the tool or during operation) is proposed considering the difference in Poisson's coefficients. A model for determining the forces acting on the perimeter of section elements was synthesized, and the corresponding formulas were obtained, which form the basis for the theory of off-center compression of rods. The direction of further research in this field is currently being planned.

Functional Prototissues Using Artificial Cells as Building Blocks and Their Biomedical Applications

AbstractThe construction of living systems from the bottom‐up helps to explore the complex processes of life and to understand their working mechanism. Prototissues, constructed using artificial cells as building blocks, mimic life systems at a high‐order tissue level, whilst artificial cells usually mimic living cells at the individual cell level. The 3D biomimetic prototissues demonstrate exceptional performances and collective functions, which reveal the working mechanisms of living tissues and hold promising potential for biomedical applications. This review systematically summarized the research progress of the field of prototissues. The engineering methods for fabricating two types of prototissues are introduced first, followed by the functions of prototissues including collective behaviors and signal communications, as well as their biomedical applications. The challenges and future trends are proposed at the end of the paper.

Automated crack localization for road safety using contextual u-net with spatial-channel feature integration

Accurate and timely crack localization is crucial for road safety and maintenance, but image processing and hand-crafted feature engineering methods, often fail to distinguish cracks from background noise under diverse lighting and surface conditions. This paper proposes a framework utilizing contextual U-Net deep learning model to automatically localize cracks in road images. The framework design considers crack localization as a task of pixel-level segmenting, and analyzing each pixel in a road image to determine if it belongs to a crack or not. The proposed U-Net model uses a robust EfficientNet encoder to capture crucial details (spatial features) and overall patterns (channel-wise features) within the road image. This balanced approach helps the model learn effectively from both individual elements and the context of the images, leading to improved crack detection. A customized hierarchical attention mechanism is designed to make U-Net model contextually adaptive to focus on relevant features at different scales and resolutions for accurately localizing road cracks that can vary widely in size and shape. The model's effectiveness is demonstrated through extensive evaluations on the benchmarked and custom-made datasets.

Using Engineering Methods (Kaizen and Micromovements Science) to Improve and Provide Evidence Regarding Microsurgical Hand Skills

ObjectiveMicrosurgical interventions involve the interaction of numerous variables, making objective analysis of skill proficiency challenging. This difficulty is even more pronounced in low-resource contexts. Continuous improvement methodologies such as Kaizen-PDCA and Micro-Movements Science can address this issue. This study aimed to demonstrate the advantages of designing and implementing microsurgical training programs using these methodologies. MethodsFollowing an extensive literature review of Kaizen-PDCA and Micro-Movements Science and under the guidance of experienced neurosurgeons and engineers, a microvascular bypass training program was developed using the human placenta. Subsequently, the training program was used to analyze and describe the process of a trainee neurosurgeon in Argentina with no prior experience in microvascular anastomosis, as the operator gained proficiency. ResultsThe trainee required 12 attempts to achieve the program goals. The longest procedural time was during the first attempt (1 h 49 min 05 s with two mistakes), while the shortest time was during the fourth attempt (53 min 29 s with three mistakes). After 12 attempts, the trainee made no mistakes, and the procedural time was reduced to 57 min 37 s. The final learning curve demonstrated a regular pattern and reached a plateau after seven attempts. ConclusionsThe training program and methodology effectively assessed, facilitated, and demonstrated the acquisition of microsurgical skills. Kaizen-PDCA and Micro-Movements Science enabled the effective use of expert experience, detailed evaluation of microsurgical procedures, and integration into a continuous improvement cycle. The program structure could also be valuable for teaching, evaluating, and enhancing similar surgical procedures.

Evaluation of the Pilot Test Effect of Steam Stimulation in Horizontal Wells of Sand Bodies 1-1308 in Bohai L Oilfield

In order to reduce the risk of large-scale thermal recovery of heavy oil in Bohai Oilfield, L Oilfield is selected to carry out horizontal well steam stimulation pilot tests. Previous studies have summarized the rationality of the injection parameters and initial production characteristics of this experiment, but the evaluation of its development effectiveness is still blank. Based on the study of the decline law of A23H well cold recovery using reservoir engineering methods, this article evaluates the initial production capacity, production increase effect, technical recoverable reserves, and throughput cycles of steam stimulation. The results show that the thermal recovery production of steam stimulation is three times that of cold recovery, which can significantly improve the production capacity of a single well. The initial daily oil production of two wells is 74m<sup>3</sup>/d and 90m<sup>3</sup>/d, respectively, which is 3.0 times and 3.6 times that of the initial production of cold recovery, exceeding the design level of the reservoir; The first round has the best yield increase effect, with an average daily oil increase of 31m<sup>3</sup>/d, and gradually decreasing thereafter. By the fifth round, it was only 13m<sup>3</sup>/d. In terms of yield increase multiples, each round can reach about 2 times; The technically recoverable reserves are 19.03 × 10<sup>4</sup>m<sup>3</sup>, requiring 8-9 rounds. Through this performance evaluation, we have strengthened our confidence in promoting the application of steam injection technology in Bohai Oilfield to achieve large-scale thermal recovery.

Comparison of Synthetic Data Generation Techniques for Control Group Survival Data in Oncology Clinical Trials: Simulation Study.

Synthetic patient data (SPD) generation for survival analysis in oncology trials holds significant potential for accelerating clinical development. Various machine learning methods, including classification and regression trees (CART), random forest (RF), Bayesian network (BN), and conditional tabular generative adversarial network (CTGAN), have been used for this purpose, but their performance in reflecting actual patient survival data remains under investigation. The aim of this study was to determine the most suitable SPD generation method for oncology trials, specifically focusing on both progression-free survival (PFS) and overall survival (OS), which are the primary evaluation end points in oncology trials. To achieve this goal, we conducted a comparative simulation of 4 generation methods, including CART, RF, BN, and the CTGAN, and the performance of each method was evaluated. Using multiple clinical trial data sets, 1000 data sets were generated by using each method for each clinical trial data set and evaluated as follows: (1) median survival time (MST) of PFS and OS; (2) hazard ratio distance (HRD), which indicates the similarity between the actual survival function and a synthetic survival function; and (3) visual analysis of Kaplan-Meier (KM) plots. Each method's ability to mimic the statistical properties of real patient data was evaluated from these multiple angles. In most simulation cases, CART demonstrated the high percentages of MSTs for synthetic data falling within the 95% CI range of the MST of the actual data. These percentages ranged from 88.8% to 98.0% for PFS and from 60.8% to 96.1% for OS. In the evaluation of HRD, CART revealed that HRD values were concentrated at approximately 0.9. Conversely, for the other methods, no consistent trend was observed for either PFS or OS. CART demonstrated better similarity than RF, in that CART caused overfitting and RF (a kind of ensemble learning approach) prevented it. In SPD generation, the statistical properties close to the actual data should be the focus, not a well-generalized prediction model. Both the BN and CTGAN methods cannot accurately reflect the statistical properties of the actual data because small data sets are not suitable. As a method for generating SPD for survival data from small data sets, such as clinical trial data, CART demonstrated to be the most effective method compared to RF, BN, and CTGAN. Additionally, it is possible to improve CART-based generation methods by incorporating feature engineering and other methods in future work.

Current Approaches and Methods to Understand Acute Ischemic Stroke Treatment Using Aspiration Thrombectomy.

Acute ischemic stroke occurs when a blood clot occludes a cerebral artery. Mechanical interventions, primarily stent retrievers and aspiration thrombectomy, are used currently for removing the occluding clot and restoring blood flow. Aspiration involves using a long catheter to traverse the cerebral vasculature to reach the blood clot, followed by application of suction through the catheter bore. Aspiration is also used in conjunction with other techniques such as stent retrievers and balloon guide catheters. Despite the wide use of aspiration, our physical understanding of the process and the causes of the failure of aspiration to retrieve cerebral clots in certain scenarios is not well understood. Experimental and computational studies can help develop the capability to provide deeper insights into the procedure and enable development of new devices and more effective treatment methods. We recapitulate the aspiration-based thrombectomy techniques in clinical practice and provide a perspective of existing engineering methods for aspiration. We articulate the current knowledge gap in the understanding of aspiration and highlight possible directions for future engineering studies to bridge this gap, help clinical translation of engineering studies, and develop new patient-specific stroke therapy.

Unique Operating Conditions Analysis of Turboshaft Engines Based on Nonlinear Integrated Model

The unique mission of the helicopter compared to the fixed-wing aircraft introduces challenges to the aerodynamic stability of the turboshaft engine. Thus, the assessment of the availability of stability margin for special operating conditions in the envelope of this equipment is required to guarantee flight safety. In this paper, a rapid nonlinear complete engine method is developed to investigate the effect of helicopter ground-effect operation and high-altitude hovering on aerodynamic stability by considering factors including inlet distortion and component matching. Research indicated that the most dangerous point of stability margin in a unique section of the flight envelope is high altitude hovering, with about a 16.7% reduction in available stability margin compared to standard ground test conditions. This paper also investigates the effects of typical helicopter ground effects on a turboshaft engine and shows that the stability margin will be reduced by a maximum of 1.43% in hovering if the influence of ground effects in the flight envelope is not considered in the design.

Synergistic Passivation on Buried Interface for Highly Efficient and Stable p-i-n Perovskite Solar Cells.

The properties of an interface at the hole transport layer (HTL)/perovskite layer are crucial for the performance and stability of perovskite solar cells (PVSCs), especially the buried interface between HTL and perovskite layer. Here, a molecular named potassium 1-trifluoroboratomethylpiperidine (3FPIP) assistant-modified perovskite bottom interface strategy is proposed to improve the charge transfer capability and balances energy level between HTL and perovskite. BF3 - in the 3FPIP molecule interacts with undercoordinated Pb2+ to passivate iodine vacancies and enhance PVSCs performance. Furthermore, the infiltration of K+ ions into perovskite molecules enhances the crystallinity and stability of perovskite. Therefore, the PVSCs with the buried interface treatment exhibit a champion performance of 24.6%. More importantly, the corresponding devices represent outstanding ambient stability, remaining at 92% of the initial efficiency after 1200h. This work provides a new method of buried interface engineering with functional group synergy.

Modulating oxygen vacancies in MXene/MoO3-x smart fiber by defect engineering for ultrahigh volumetric energy density supercapacitors and wearable SERS sensors

MXene fibers are expected to accelerate the sustainable development of intelligent era as multi-functional smart fibers. However, MXene fibers have serious obstacles when used for multi-functional applications. Here, we fabricated a multi-functional MXene fiber by incorporating MoO3-x nanobelts with abundant oxygen vacancies via wet spinning and defect engineering methods. Benefiting from the presence of abundant oxygen vacancies, the produced MXene/MoO3-x fibers not only provide more active sites to interact with electrolyte ions, but also substantially augment the rate capacities of fibers due to the enlarged distances between MXene flakes. Thus, the assembled fiber-based supercapacitor (SCs) realized high capacitance of 869.1F cm−3 at 0.5 A cm−3, good volumetric energy density of 77.3 mWh cm−3 at a power density of 204.8 mW cm−3 and long-term cycling stability along with 86.4 % capacity retention after 5000 cycles. Moreover, due to the narrowed bandgap of MoO3-x produced by defect engineering enhanced the charge transfer between the fiber and molecules, MXene/MoO3-x fiber also demonstrated a low limit of detection and reliable biomarkers detection in artificial sweat. These findings provide the potential application of MXene-based fibers in multi-functional devices.

Unlocking large memory windows and 16-level data per cell memory operations in hafnia-based ferroelectric transistors

Ferroelectric transistors based on hafnia-based ferroelectrics exhibit tremendous potential as next-generation memories owing to their high-speed operation and low power consumption. Nevertheless, these transistors face limitations in terms of memory window, which directly affects their ability to support multilevel characteristics in memory devices. Furthermore, the absence of an efficient operational technique capable of achieving multilevel characteristics has hindered their development. To address these challenges, we present a gate stack engineering method and an efficient operational approach for ferroelectric transistors to achieve 16-level data per cell operation. By using the suggested engineering method, we demonstrate the attainment of a substantial memory window of 10 V without increasing the device area. Additionally, we propose a displacement current control method, facilitating one-shot programming to the desired state. Remarkably, we suggest the compatibility of these proposed methods with three-dimensional (3D) structures. This study underscores the potential of ferroelectric transistors for next-generation 3D memory applications.

3D bioprinting platform development for high-throughput cancer organoid models construction and drug evaluation

The evaluation of anti-tumor drugs is critical for their development and clinical guidance. Tumor organoid models are gaining increased attention due to their ability to better mimic real tumor tissues, as well as lower time and economic costs, which makes up for the shortcomings of cell lines and xenograft models. However, current tumor organoid cultures based on the Matrigel have limitations in matching with high-throughput engineering methods due to slow gelation and low mechanical strength. Here, we present a novel composite bioink for culturing colorectal cancer organoids that provides an environment close to real tissue growth conditions and exhibits excellent photocrosslinking properties for rapid gel formation. Most importantly, the tumor organoids viability in the composite bioink after printing was as high as 97%, which also kept multicellular polar structures consistent with traditional culture methods in the Matrigel. Using 3D bioprinting with this composite bioink loaded with organoids, we demonstrated the feasibility of this drug evaluation model by validating it with clinically used colorectal cancer treatment drugs. Our results suggested that the composite bioink could effectively cultivate tumor organoids using 3D bioprinting, which had the potential to replace less reliable manual operations in promoting the application of tumor organoids in drug development and clinical guidance.

Mechanism-Guided Computational Design Drives meso-Diaminopimelate Dehydrogenase to Efficient Synthesis of Aromatic d-amino Acids.

Aromatic d-amino acids (d-AAs) play a pivotal role as important chiral building blocks and key intermediates in fine chemical and drug synthesis. Meso-diaminopimelate dehydrogenase (DAPDH) serves as an excellent biocatalyst in the synthesis of d-AAs and their derivatives. However, its strict substrate specificity and the lack of efficient engineering methods have hindered its widespread application. Therefore, this study aims to elucidate the catalytic mechanism underlying DAPDH from Proteus vulgaris (PvDAPDH) through the examination of its crystallographic structure, computational simulations of potential energies and molecular dynamics simulations, and site-directed mutagenesis. Mechanism-guided computational design showed that the optimal mutant PvDAPDH-M3 increased specific activity and catalytic efficiency (kcat/Km) for aromatic keto acids up to 124-fold and 92.4-fold, respectively, compared to that of the wild type. Additionally, it expanded the substrate scope to 10 aromatic keto acid substrates. Finally, six high-value-added aromatic d-AAs and their derivatives were synthesized using a one-pot three-enzyme cascade reaction, exhibiting a good conversion rate ranging from 32 to 84% and excellent stereoselectivity (enantiomeric excess >99%). These findings provide a potential synthetic pathway for the green industrial production of aromatic d-AAs.