scholarly journals The N-Terminal β-Sheet of the Hyperthermophilic Endoglucanase from Pyrococcus horikoshii Is Critical for Thermostability

2012 ◽  
Vol 78 (9) ◽  
pp. 3059-3067 ◽  
Author(s):  
Trent C. Yang ◽  
Steve Legault ◽  
Emery A. Kayiranga ◽  
Jyothi Kumaran ◽  
Kazuhiko Ishikawa ◽  
...  
Keyword(s):  
Triosephosphate Isomerase ◽  
Industrial Applications ◽  
Crystalline Cellulose ◽  
Bioinformatics Analyses ◽  
Pyrococcus Horikoshii ◽  
Content Type ◽  
Enzyme Thermostability ◽  
Tim Barrel ◽  
Key Residues ◽  
Β Sheet

ABSTRACTThe β-1,4-endoglucanase (EC 3.2.1.4) from the hyperthermophilic archaeonPyrococcus horikoshii(EGPh) has strong hydrolyzing activity toward crystalline cellulose. When EGPh is used in combination with β-glucosidase (EC 3.2.1.21), cellulose is completely hydrolyzed to glucose at high temperature, suggesting great potential for EGPh in bioethanol industrial applications. The crystal structure of EGPh shows a triosephosphate isomerase (TIM) (β/α)8-barrel fold with an N-terminal antiparallel β-sheet at the opposite side of the active site and a very short C-terminal sequence outside of the barrel structure. We describe here the function of the peripheral sequences outside of the TIM barrel core structure. Sequential deletions were performed from both N and C termini. The activity, thermostability, and pH stability of the expressed mutants were assessed and compared to the wild-type EGPh enzyme. Our results demonstrate that the TIM barrel core is essential for enzyme activity and that the N-terminal β-sheet is critical for enzyme thermostability. Bioinformatics analyses identified potential key residues which may contribute to enzyme hyperthermostability.

scholarly journals Activity and Thermostability of GH5 Endoglucanase Chimeras from Mesophilic and Thermophilic Parents

2018 ◽  
Vol 85 (5) ◽  
Author(s):  
Fei Zheng ◽  
Josh V. Vermaas ◽  
Jie Zheng ◽  
Yuan Wang ◽  
Tao Tu ◽  
...  
Keyword(s):  
Thermal Properties ◽  
Rational Design ◽  
Catalytic Efficiency ◽  
Md Simulations ◽  
Industrial Applications ◽  
Glycoside Hydrolase Family ◽  
Essential Property ◽  
Content Type ◽  
Tim Barrel ◽  
Hybrid Enzymes

ABSTRACT Cellulases from glycoside hydrolase family 5 (GH5) are key endoglucanase enzymes in the degradation of diverse polysaccharide substrates and are used in industrial enzyme cocktails to break down biomass. The GH5 family shares a canonical (βα)8-barrel structure, where each (βα) module is essential for the enzyme’s stability and activity. Despite their shared topology, the thermostability of GH5 endoglucanase enzymes can vary significantly, and highly thermostable variants are often sought for industrial applications. Based on the previously characterized thermophilic GH5 endoglucanase Egl5A from Talaromyces emersonii (TeEgl5A), which has an optimal temperature of 90°C, we created 10 hybrid enzymes with elements of the mesophilic endoglucanase Cel5 from Stegonsporium opalus (SoCel5) to determine which elements are responsible for enhanced thermostability. Five of the expressed hybrid enzymes exhibit enzyme activity. Two of these hybrids exhibited pronounced increases in the temperature optimum (10 and 20°C), the temperature at which the protein lost 50% of its activity (T50) (15 and 19°C), and the melting temperature (Tm) (16.5 and 22.9°C) and extended half-lives (t1/2) (∼240- and 650-fold at 55°C) relative to the values for the mesophilic parent enzyme and demonstrated improved catalytic efficiency on selected substrates. The successful hybridization strategies were validated experimentally in another GH5 endoglucanase, Cel5 from Aspergillus niger (AnCel5), which demonstrated a similar increase in thermostability. Based on molecular dynamics (MD) simulations of both the SoCel5 and TeEgl5A parent enzymes and their hybrids, we hypothesize that improved hydrophobic packing of the interface between α2 and α3 is the primary mechanism by which the hybrid enzymes increase their thermostability relative to that of the mesophilic parent SoCel5. IMPORTANCE Thermal stability is an essential property of enzymes in many industrial biotechnological applications, as high temperatures improve bioreactor throughput. Many protein engineering approaches, such as rational design and directed evolution, have been employed to improve the thermal properties of mesophilic enzymes. Structure-based recombination has also been used to fuse TIM barrel fragments, and even fragments from unrelated folds, to generate new structures. However, little research has been done on GH5 endoglucanases. In this study, two GH5 endoglucanases exhibiting TIM barrel structure, SoCel5 and TeEgl5A, with different thermal properties, were hybridized to study the roles of different (βα) motifs. This work illustrates the role that structure-guided recombination can play in helping to identify sequence function relationships within GH5 enzymes by supplementing natural diversity with synthetic diversity.

Purification and Characterization of Natural Solid-Substrate Degrading and Alcohol Producing Hyperthermostable Alkaline Amylase from Bacillus cereus (sm-sr14)

2020 ◽  
Vol 21 (9) ◽  
pp. 872-881
Author(s):  
Sumit Sahoo ◽  
Sudipta Roy ◽  
Dipannita Santra ◽  
Sayantani Maiti ◽  
Sonali Roul ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Main Property ◽  
Solid Substrate ◽  
Industrial Applications ◽  
Starch Degradation ◽  
Significant Similarity ◽  
Alcohol Production ◽  
Cazy Database ◽  
Tim Barrel ◽  
Alkaline Amylase

Objective: Amylases enzymes hydrolyze starch molecules to produce diverse products including dextrins, and progressively smaller polymers. These include glucose units linked through α-1- 1, α-1-4, α-1-6, glycosidic bonds. Methods: This enzyme carrying an (α /β) 8 or TIM barrel structure is also produced containing the catalytic site residues. These groups of enzymes possess four conserved regions in their primary sequence. In the Carbohydrate-Degrading Enzyme (CAZy) database, α-amylases are classified into different Glycoside Hydrolase Families (GHF) based on their amino acid sequence. The present objective was to study one such enzyme based on its molecular characterization after purification in our laboratory. Its main property of solid-natural starch degradation was extensively investigated for its pharmaceutical/ industrial applications. Results: Amylase producing bacteria Bacillus cereus sm-sr14 (Accession no. KM251578.1) was purified to homogeneity on a Seralose 6B-150 gel-matrix and gave a single peak during HPLC. MALDITOF mass-spectrometry with bioinformatics studies revealed its significant similarity to α/β hydrolase family. The enzyme showed an efficient application; favourable Km, Vmax and Kcat during the catalysis of different natural solid starch materials. Analysis for hydrolytic product showed that this enzyme can be classified as the exo-amylase asit produced a significant amount of glucose. Conclusion: Besides the purified enzyme, the present organism Bacillus cereus sm-sr14 could degrade natural solid starch materials like potato and rice up to the application level in the pharmaceutical/ industrial field for alcohol production.

scholarly journals Two β-glucuronosyltransferases involved in the biosynthesis of type II arabinogalactans function in mucilage polysaccharide matrix organization in Arabidopsis thaliana

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Oyeyemi O. Ajayi ◽  
Michael A. Held ◽  
Allan M. Showalter
Keyword(s):  
Arabidopsis Thaliana ◽  
Seed Size ◽  
Seed Coat ◽  
Glucuronic Acid ◽  
Industrial Applications ◽  
Crystalline Cellulose ◽  
Sugar Residue ◽  
Composition Analysis ◽  
Cellulose Content ◽  
Type Ii

Abstract Background Arabinogalactan-proteins (AGPs) are heavily glycosylated with type II arabinogalactan (AG) polysaccharides attached to hydroxyproline residues in their protein backbone. Type II AGs are necessary for plant growth and critically important for the establishment of normal cellular functions. Despite the importance of type II AGs in plant development, our understanding of the underlying role of these glycans/sugar residues in mucilage formation and seed coat epidermal cell development is poorly understood and far from complete. One such sugar residue is the glucuronic acid residues of AGPs that are transferred onto AGP glycans by the action of β-glucuronosyltransferase genes/enzymes. Results Here, we have characterized two β-glucuronosyltransferase genes, GLCAT14A and GLCAT14C, that are involved in the transfer of β-glucuronic acid (GlcA) to type II AGs. Using a reverse genetics approach, we observed that glcat14a-1 mutants displayed subtle alterations in mucilage pectin homogalacturonan (HG) compared to wild type (WT), while glcat14a-1glcat14c-1 mutants displayed much more severe mucilage phenotypes, including loss of adherent mucilage and significant alterations in cellulose ray formation and seed coat morphology. Monosaccharide composition analysis showed significant alterations in the sugar amounts of glcat14a-1glcat14c-1 mutants relative to WT in the adherent and non-adherent seed mucilage. Also, a reduction in total mucilage content was observed in glcat14a-1glcat14c-1 mutants relative to WT. In addition, glcat14a-1glcat14c-1 mutants showed defects in pectin formation, calcium content and the degree of pectin methyl-esterification (DM) as well as reductions in crystalline cellulose content and seed size. Conclusions These results raise important questions regarding cell wall polymer interactions and organization during mucilage formation. We propose that the enzymatic activities of GLCAT14A and GLCAT14C play partially redundant roles and are required for the organization of the mucilage matrix and seed size in Arabidopsis thaliana. This work brings us a step closer towards identifying potential gene targets for engineering plant cell walls for industrial applications.

Wearable sensors bring new benefits to continuous medical monitoring, real time physical activity assessment, baby monitoring and industrial applications

Sensor Review ◽  
2015 ◽  
Vol 35 (2) ◽  
pp. 141-145 ◽  
Author(s):  
Richard Bloss
Keyword(s):  
Real Time ◽  
Continuous Monitoring ◽  
Medical Condition ◽  
Wearable Sensors ◽  
Medical Personnel ◽  
Industrial Applications ◽  
Medical Monitoring ◽  
Content Type ◽  
Doctor's Office ◽  
Monitoring Technologies

Purpose – The purpose of this paper is to review the recent advancements in the development of wearable sensors which can continuously monitor critical medical, assess athletic activity, watch babies and serve industrial applications. Design/methodology/approach – The paper presents an in-depth review of a number of developments in wearable sensing and monitoring technologies for medical, athletic and industrial applications. Researchers and companies around the world were contacted to discuss their direction and progress in this field of medical condition and industrial monitoring, as well as discussions with medical personnel on the perceived benefits of such technology. Findings – Dramatic progress is being made in continuous monitoring of many important body functions that indicate critical medical conditions that can be life-threatening, contribute to blindness or access activity. In the industrial arena, wearable devices bring remote monitoring to a new level. Practical implications – Doctors will be able to replace one-off tests with continuous monitoring that provides a much better continuous real-time “view” into the patient’s conditions. Wearable monitors will help provide much better medical care in the future. Industrial managers and others will be able to monitor and supervise remotely. Originality/value – An expert insight into advancements in medical condition monitoring that replaces the one-time “finger prick” type testing only performed in the doctor’s office. It is also a look at how wearable monitoring is greatly improved and serving athletics, the industry and parents.

scholarly journals Value-driven engineering of E-maintenance platforms

2014 ◽  
Vol 25 (4) ◽  
pp. 568-598 ◽  
Author(s):  
Marco Macchi ◽  
Adolfo Crespo Márquez ◽  
Maria Holgado ◽  
Luca Fumagalli ◽  
Luis Barberá Martínez
Keyword(s):  
Information And Communication Technologies ◽  
Cost Benefit ◽  
Communication Technologies ◽  
Industrial Applications ◽  
Maintenance Management ◽  
Content Type ◽  
Positive Effects ◽  
A Value ◽  
New Information ◽  
Information And Communication

Purpose – The purpose of this paper is to propose a methodology for the engineering of E-maintenance platforms that is based on a value-driven approach. Design/methodology/approach – The methodology assumes that a value-driven engineering approach would help foster technological innovation for maintenance management. Indeed, value-driven engineering could be easily adopted at the business level, with subsequent positive effects on the industrial applications of new information and communication technologies solutions. Findings – The methodology combines a value-driven approach with the engineering in the maintenance scope. The methodology is tested in a manufacturing case to prove its potential to support the engineering of E-maintenance solutions. In particular, the case study concerns the investment in E-maintenance solutions developed in the framework of a Supervisory Control and Data Acquisition system originally implemented for production purposes. Originality/value – Based on literature research, the paper presents a methodology that is implemented considering three different approaches (business theories, value-driven engineering and maintenance management). The combination of these approaches is novel and overcomes the traditional view of maintenance as an issue evaluated from a cost-benefit perspective.

The role of robotics in additive manufacturing: review of the AM processes and introduction of an intelligent system

2021 ◽  
Vol ahead-of-print (ahead-of-print) ◽  
Author(s):  
J. Norberto Pires ◽  
Amin S. Azar ◽  
Filipe Nogueira ◽  
Carlos Ye Zhu ◽  
Ricardo Branco ◽  
...  
Keyword(s):  
Additive Manufacturing ◽  
Computer Aided Design ◽  
Intelligent System ◽  
Material Selection ◽  
High Strength Steels ◽  
Industrial Applications ◽  
Layer By Layer ◽  
Content Type ◽  
Functional Components

Purpose Additive manufacturing (AM) is a rapidly evolving manufacturing process, which refers to a set of technologies that add materials layer-by-layer to create functional components. AM technologies have received an enormous attention from both academia and industry, and they are being successfully used in various applications, such as rapid prototyping, tooling, direct manufacturing and repair, among others. AM does not necessarily imply building parts, as it also refers to innovation in materials, system and part designs, novel combination of properties and interplay between systems and materials. The most exciting features of AM are related to the development of radically new systems and materials that can be used in advanced products with the aim of reducing costs, manufacturing difficulties, weight, waste and energy consumption. It is essential to develop an advanced production system that assists the user through the process, from the computer-aided design model to functional components. The challenges faced in the research and development and operational phase of producing those parts include requiring the capacity to simulate and observe the building process and, more importantly, being able to introduce the production changes in a real-time fashion. This paper aims to review the role of robotics in various AM technologies to underline its importance, followed by an introduction of a novel and intelligent system for directed energy deposition (DED) technology. Design/methodology/approach AM presents intrinsic advantages when compared to the conventional processes. Nevertheless, its industrial integration remains as a challenge due to equipment and process complexities. DED technologies are among the most sophisticated concepts that have the potential of transforming the current material processing practices. Findings The objective of this paper is identifying the fundamental features of an intelligent DED platform, capable of handling the science and operational aspects of the advanced AM applications. Consequently, we introduce and discuss a novel robotic AM system, designed for processing metals and alloys such as aluminium alloys, high-strength steels, stainless steels, titanium alloys, magnesium alloys, nickel-based superalloys and other metallic alloys for various applications. A few demonstrators are presented and briefly discussed, to present the usefulness of the introduced system and underlying concept. The main design objective of the presented intelligent robotic AM system is to implement a design-and-produce strategy. This means that the system should allow the user to focus on the knowledge-based tasks, e.g. the tasks of designing the part, material selection, simulating the deposition process and anticipating the metallurgical properties of the final part, as the rest would be handled automatically. Research limitations/implications This paper reviews a few AM technologies, where robotics is a central part of the process, such as vat photopolymerization, material jetting, binder jetting, material extrusion, powder bed fusion, DED and sheet lamination. This paper aims to influence the development of robot-based AM systems for industrial applications such as part production, automotive, medical, aerospace and defence sectors. Originality/value The presented intelligent system is an original development that is designed and built by the co-authors J. Norberto Pires, Amin S. Azar and Trayana Tankova.

Fast and accurate detection of surface defect based on improved YOLOv4

2021 ◽  
Vol ahead-of-print (ahead-of-print) ◽  
Author(s):  
Jiawei Lian ◽  
Junhong He ◽  
Yun Niu ◽  
Tianze Wang
Keyword(s):  
Feature Extraction ◽  
Real Time ◽  
Surface Defect ◽  
Steel Ingot ◽  
Industrial Applications ◽  
Data Sets ◽  
Data Set ◽  
Processing Technologies ◽  
Content Type ◽  
Public Data

Purpose The current popular image processing technologies based on convolutional neural network have the characteristics of large computation, high storage cost and low accuracy for tiny defect detection, which is contrary to the high real-time and accuracy, limited computing resources and storage required by industrial applications. Therefore, an improved YOLOv4 named as YOLOv4-Defect is proposed aim to solve the above problems. Design/methodology/approach On the one hand, this study performs multi-dimensional compression processing on the feature extraction network of YOLOv4 to simplify the model and improve the feature extraction ability of the model through knowledge distillation. On the other hand, a prediction scale with more detailed receptive field is added to optimize the model structure, which can improve the detection performance for tiny defects. Findings The effectiveness of the method is verified by public data sets NEU-CLS and DAGM 2007, and the steel ingot data set collected in the actual industrial field. The experimental results demonstrated that the proposed YOLOv4-Defect method can greatly improve the recognition efficiency and accuracy and reduce the size and computation consumption of the model. Originality/value This paper proposed an improved YOLOv4 named as YOLOv4-Defect for the detection of surface defect, which is conducive to application in various industrial scenarios with limited storage and computing resources, and meets the requirements of high real-time and precision.

A study on tribological behaviour and analysis of ZnO reinforced AA6061 matrix composites fabricated by stir casting route

2021 ◽  
Vol ahead-of-print (ahead-of-print) ◽  
Author(s):  
Sakthi Sadhasivam RM ◽  
Ramanathan K. ◽  
Bhuvaneswari B.V. ◽  
Raja R.
Keyword(s):  
Wear Rate ◽  
Sliding Wear ◽  
Stir Casting ◽  
Industrial Applications ◽  
Wear Behaviour ◽  
Matrix Composites ◽  
Tribological Behaviour ◽  
Content Type ◽  
Zno Particles ◽  
The Matrix

Purpose The most promising replacements for the industrial applications are particle reinforced metal matrix composites because of their good and combined mechanical properties. Currently, the need of matrix materials for industrial applications is widely satisfied by aluminium alloys. The purpose of this paper is to evaluate the tribological behaviour of the zinc oxide (ZnO) particles reinforced AA6061 composites prepared by stir casting route. Design/methodology/approach In this study, AA6061 aluminium alloy matrix reinforced with varying weight percentages (3%, 4.5% and 6%) of ZnO particles, including monolithic AA6061 alloy samples, is cast by the most economical fabrication method, called stir casting. The prepared sample was subjected to X-ray photoelectron spectroscopy (XPS) analysis, experimental density measurement by Archimedian principle and theoretical density by rule of mixture and hardness test to investigate mechanical property. The dry sliding wear behaviour of the composites was investigated using pin-on-disc tribometer with various applied loads of 15 and 20 N, with constant sliding velocity and distance. The wear rate, coefficient of friction (COF) and worn surfaces of the composite specimens and their effects were also investigated in this work. Findings XPS results confirm the homogeneous distribution of ZnO microparticles in the Al matrix. The Vickers hardness result reveals that higher ZnO reinforced (6%) sample have 34.4% higher values of HV than the monolithic aluminium sample. The sliding wear tests similarly show that increasing the weight percentage of ZnO particles leads to a reduced wear rate and COF of 30.01% and 26.32% lower than unreinforced alloy for 15 N and 36.35% and 25% for 20 N applied load. From the worn surface morphological studies, it was evidently noticed that ZnO particles dispersed throughout the matrix and it had strong bonding between the reinforcement and the matrix, which significantly reduced the plastic deformation of the surfaces. Originality/value The uniqueness of this work is to use the reinforcement of ZnO particles with AA6061 matrix and preparing by stir casting route and to study and analyse the physical, hardness and tribological behaviour of the composite materials.

A 12-pulse rectifier with passive harmonic reduction based on UIPT in more electric aircrafts

Circuit World ◽  
2021 ◽  
Vol ahead-of-print (ahead-of-print) ◽  
Author(s):  
Rohollah Abdollahi
Keyword(s):  
Power Factor ◽  
Harmonic Distortion ◽  
Cost Savings ◽  
Cost Effective ◽  
Industrial Applications ◽  
Input Current ◽  
New Combination ◽  
Content Type ◽  
Harmonic Reduction ◽  
Simulation Results

Purpose The purpose of this paper is to provide a T autotransformer based 12-pulse rectifier with passive harmonic reduction in more electric aircraft applications. The T autotransformer uses only two main windings which result in volume, space, size, weight and cost savings. Also, the proposed unconventional inter-phase transformer (UIPT) with a lower kVA rating (about 2.6% of the load power) compared to the conventional inter-phase transformer results in a more harmonic reduction. Design/methodology/approach To increase rating and reduce the cost and complexity of a multi-pulse rectifier, it is well known that the pulse number must be increased. In some practical cases, a 12-pulse rectifier (12PR) is suggested as a good solution considering its simple structure and low weight. But the 12PR cannot technically meet the standards of harmonic distortion requirements for some industrial applications, and therefore, they must be used with output filters. In this paper, a 12PR is suggested, which consists of a T autotransformer 12PR and a passive harmonic reduction (PHR) based on the UIPT at direct current (DC) link. Findings To show the advantage of this new combination over other solutions, simulation results are used, and then, a prototype is implemented to evaluate and verify the simulation results. The simulation and experimental test results show that the input current total harmonic distortion (THD) of the suggested 12PR with a PHR based on UIPT is less than 5%, which meets the IEEE 519 requirements. Also, it is shown that in comparison with other solutions, it is cost effective, and at the same time, its power factor is near unity, and its rating is 29.92% of the load rating. Therefore, it is obvious that the proposed rectifier is a practical solution for more electric aircrafts. Originality/value The contributions of this paper are summarized as follows. The suggested design uses a retrofit T autotransformer, which meets all technical constraints, and in comparison, with other options, has less rating, weight, volume and cost. In the suggested rectifier, a PHR based on UIPT at its dc link of 12PR is used, which has good technical capabilities and lower ratings. In the PHR based on UIPT, an IPT is used, which has an additional secondary winding and four diodes. This solution leads to a reduction in input current THD and conduction losses of diodes. In full load conditions, the input line current THD and power factor are 4% and 0.99, respectively. The THD is less than 5%, which satisfies IEEE-519 and DO-160G requirements.

Experimental study of pin surface precision on lubrication behavior of pin–bush pairs in industrial roller chains

2021 ◽  
Vol ahead-of-print (ahead-of-print) ◽  
Author(s):  
Ansheng Zhang ◽  
Mingyu Zhang ◽  
Jing Wang ◽  
Jianjun Zhang ◽  
Zhaohua Shang ◽  
...  
Keyword(s):  
Experimental Study ◽  
Design Methodology ◽  
Industrial Applications ◽  
Experimental Basis ◽  
Reciprocating Motion ◽  
Content Type ◽  
Oil Supply ◽  
Machining Errors ◽  
Surface Precision ◽  
Straightness Error

Purpose The purpose of this paper is to study the influence of surface precision on the lubrication state of the roller chain under adequate and rare oil supply conditions, respectively. Design/methodology/approach The straightness error and roughness error of the pin generatrices were measured and the influence of surface precision on the lubrication behavior under steady state and reciprocating motion was studied through optical interference experiments. Findings The lubrication state is strongly influenced by the surface precision of the roller surface both under adequate oil supply and rare oil supply conditions. Originality/value In industrial applications, the machining errors of parts cannot be completely eliminated. Studying the influence of the surface precision on the lubrication behavior of pin–bush pairs can provide the experimental basis for the optimal design of the bush roller chains.

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