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Heterologous Expression of a Thermostable α-Galactosidase from Parageobacillus thermoglucosidasius Isolated from the Lignocellulolytic Microbial Consortium TMC7

  • Wang, Yi;Wang, Chen;Chen, Yonglun;Cui, MingYu;Wang, Qiong;Guo, Peng
    • Journal of Microbiology and Biotechnology
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    • v.32 no.6
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    • pp.749-760
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    • 2022
  • α-Galactosidase is a debranching enzyme widely used in the food, feed, paper, and pharmaceuticals industries and plays an important role in hemicellulose degradation. Here, T26, an aerobic bacterial strain with thermostable α-galactosidase activity, was isolated from laboratory-preserved lignocellulolytic microbial consortium TMC7, and identified as Parageobacillus thermoglucosidasius. The α-galactosidase, called T26GAL and derived from the T26 culture supernatant, exhibited a maximum enzyme activity of 0.4976 IU/ml when cultured at 60℃ and 180 rpm for 2 days. Bioinformatics analysis revealed that the α-galactosidase T26GAL belongs to the GH36 family. Subsequently, the pET-26 vector was used for the heterologous expression of the T26 α-galactosidase gene in Escherichia coli BL21 (DE3). The optimum pH for α-galactosidase T26GAL was determined to be 8.0, while the optimum temperature was 60℃. In addition, T26GAL demonstrated a remarkable thermostability with more than 93% enzyme activity, even at a high temperature of 90℃. Furthermore, Ca2+ and Mg2+ promoted the activity of T26GAL while Zn2+ and Cu2+ inhibited it. The substrate specificity studies revealed that T26GAL efficiently degraded raffinose, stachyose, and guar gum, but not locust bean gum. This study thus facilitated the discovery of an effective heat-resistant α-galactosidase with potent industrial application. Meanwhile, as part of our research on lignocellulose degradation by a microbial consortium, the present work provides an important basis for encouraging further investigation into this enzyme complex.

Analysis of the buckling failure of bedding slope based on monitoring data - a model test study

  • Zhang, Qian;Hu, Jie;Gao, Yang;Du, Yanliang;Li, Liping;Liu, Hongliang;Sun, Shangqu
    • Geomechanics and Engineering
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    • v.28 no.4
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    • pp.335-346
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    • 2022
  • Buckling failure is a typical slope instability mode that should be paid more attention to. It is difficult to provide systematic guidance for the monitoring and management of such slopes due to unclear mechanism. Here we examine buckling failure as the potential instability mode for a slope above a railway tunnel in southwest China. A comprehensive model test system was developed that can be used to conduct buckling failure experiments. The displacement, stress, and strain of the slope were monitored to document the evolution of buckling failure during the experiment. Monitoring data reveal the deformation and stress characteristics of the slope with different slipping mass thicknesses and under different top loads. The test results show that the slipping mass is the main subject of the top load and is the key object of monitoring. Displacement and stress precede buckling failure, so maybe useful predictors of impending failure. However, the response of the stress variation is earlier than displacement variation during the failure process. It is also necessary to monitor the bedrock near the slip face because its stress evolution plays an important role in the early prediction of instability. The position near the slope foot is most prone to buckling failure, so it should be closely monitored.

A murine periodontitis model using coaggregation between human pathogens and a predominant mouse oral commensal bacterium

  • Liu, Mengmeng;Choi, Youngnim
    • Journal of Periodontal and Implant Science
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    • v.52 no.2
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    • pp.141-154
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    • 2022
  • Purpose: C57BL/6 mice, which are among the most common backgrounds for genetically engineered mice, are resistant to the induction of periodontitis by oral infection with periodontal pathogens. This study aimed to develop a periodontitis model in C57BL/6 mice using coaggregation between human pathogens and the mouse oral commensal Streptococcus danieliae (Sd). Methods: The abilities of Porphyromonas gingivalis ATCC 33277 (Pg33277), P. gingivalis ATCC 49417 (Pg49417), P. gingivalis KUMC-P4 (PgP4), Fusobacterium nucleatum subsp. nucleatum ATCC 25586 (Fnn), and F. nucleatum subsp. animalis KCOM 1280 (Fna) to coaggregate with Sd were tested by a sedimentation assay. The Sd-noncoaggregating Pg33277 and 2 Sd-coaggregating strains, PgP4 and Fna, were chosen for animal experiments. Eighty C57BL/6 mice received oral gavage with Sd once and subsequently received vehicle alone (sham), Fna, Pg33277, PgP4, or Fna+PgP4 6 times at 2-day intervals. Mice were evaluated at 5 or 8 weeks after the first gavage of human strains. Results: Fnn, Fna, and PgP4 efficiently coaggregated with Sd, but Pg33277 and Pg49417 did not. Alveolar bone loss was significantly higher in the PgP4 group at both time points (weeks 5 and 8) and in all experimental groups at week 8 compared with the sham group. The PgP4 group presented greater alveolar bone loss than the other experimental groups at both time points. A higher degree of alveolar bone loss accompanied higher bacterial loads in the oral cavity, the invasion of not only PgP4 but also Sd and Fna, and the serum antibody responses to these bacteria. Conclusions: Periodontitis was successfully induced in C57BL/6 mice by oral infection with a P. gingivalis strain that persists in the oral cavity through coaggregation with a mouse oral commensal bacterium. This new model will be useful for studying the role of human oral bacteria-host interactions in periodontitis using genetically engineered mice.

Study on the progressive collapse resistance of CP-FBSP connections in L-CFST frame structure

  • Xiong, Qingqing;Wu, Wenbo;Zhang, Wang;Chen, Zhihua;Liu, Hongbo;Su, Tiancheng
    • Steel and Composite Structures
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    • v.44 no.3
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    • pp.437-450
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    • 2022
  • When the vertical load-bearing members in high-rise structures fail locally, the beam-column joints play an important role in the redistribution of the internal forces. In this paper, a static laboratory test of three full-scale flush flange beam-reinforced connections with side and cover plates (CP-FBSP connection) with double half-span steel beams and single L-shaped columns composed of concrete-filled steel tubes (L-CFST columns) was conducted. The influence of the side plate width and cover plate thickness on the progressive collapse resistance of the substructure was thoroughly analyzed. The failure mode, vertical force-displacement curves, strain variation, reaction force of the pin support and development of internal force in the section with the assumed plastic hinge were discussed. Then, through the verified finite element model, the corresponding analyses of the thickness and length of the side plates, the connecting length between the steel beam flange and cover plate, and the vertical-force eccentricity were carried out. The results show that the failure of all the specimens occurred through the cracking of the beam flange or the cover plate, and the beam chord rotations measured by the test were all greater than 0.085 rad. Increasing the length, thickness and width of the side plates slightly reduced the progressive collapse resistance of the substructures. The vertical-force eccentricity along the beam length reduced the progressive collapse resistance of the substructure. An increase in the connecting length between the beam flange and cover plate can significantly improve the progressive collapse resistance of substructures.

In Vitro Evaluation of Two Novel Antimalarial Derivatives of SKM13: SKM13-MeO and SKM13-F

  • Thuy-Tien Thi Trinh;Young-ah Kim;Hyelee Hong;Linh Thi Thuy Le;Hayoung Jang;Soon-Ai Kim;Hyun Park;Hak Sung Kim;Seon-Ju Yeo
    • Parasites, Hosts and Diseases
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    • v.60 no.6
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    • pp.401-407
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    • 2022
  • Antimalarial drugs play an important role in the control and treatment of malaria, a deadly disease caused by the protozoan parasite Plasmodium spp. The development of novel antimalarial agents effective against drug-resistant malarial parasites is urgently needed. The novel derivatives, SKM13-MeO and SKM13-F, were designed based on an SKM13 template by replacing the phenyl group with electron-donating (-OMe) or electron-withdrawing groups (-F), respectively, to reverse the electron density. A colorimetric assay was used to quantify cytotoxicity, and in vitro inhibition assays were performed on 3 different blood stages (ring, trophozoite, and schizonts) of P. falciparum 3D7 and the ring/mixed stage of D6 strain after synchronization. The in vitro cytotoxicity analysis showed that 2 new SKM13 derivatives reduced the cytotoxicity of the SKM13 template. SKM13 maintained the IC50 at the ring and trophozoite stages but not at the schizont stage. The IC50 values for both the trophozoite stage of P. falciparum 3D7 and ring/mixed stages of D6 demonstrated that 2 SKM13 derivatives had decreased antimalarial efficacy, particularly for the SKM13-F derivative. SKM13 may be comparably effective in ring and trophozoite, and electron-donating groups (-OMe) may be better maintain the antimalarial activity than electron-withdrawing groups (-F) in SKM13 modification.

Formulation and evaluation a finite element model for free vibration and buckling behaviours of functionally graded porous (FGP) beams

  • Abdelhak Mesbah;Zakaria Belabed;Khaled Amara;Abdelouahed Tounsi;Abdelmoumen A. Bousahla;Fouad Bourada
    • Structural Engineering and Mechanics
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    • v.86 no.3
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    • pp.291-309
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    • 2023
  • This paper addresses the finite element modeling of functionally graded porous (FGP) beams for free vibration and buckling behaviour cases. The formulated finite element is based on simple and efficient higher order shear deformation theory. The key feature of this formulation is that it deals with Euler-Bernoulli beam theory with only three unknowns without requiring any shear correction factor. In fact, the presented two-noded beam element has three degrees of freedom per node, and the discrete model guarantees the interelement continuity by using both C0 and C1 continuities for the displacement field and its first derivative shape functions, respectively. The weak form of the governing equations is obtained from the Hamilton principle of FGP beams to generate the elementary stiffness, geometric, and mass matrices. By deploying the isoparametric coordinate system, the derived elementary matrices are computed using the Gauss quadrature rule. To overcome the shear-locking phenomenon, the reduced integration technique is used for the shear strain energy. Furthermore, the effect of porosity distribution patterns on the free vibration and buckling behaviours of porous functionally graded beams in various parameters is investigated. The obtained results extend and improve those predicted previously by alternative existing theories, in which significant parameters such as material distribution, geometrical configuration, boundary conditions, and porosity distributions are considered and discussed in detailed numerical comparisons. Determining the impacts of these parameters on natural frequencies and critical buckling loads play an essential role in the manufacturing process of such materials and their related mechanical modeling in aerospace, nuclear, civil, and other structures.

Identification of 1,3,6,8-Tetrahydroxynaphthalene Synthase (ThnA) from Nocardia sp. CS682

  • Purna Bahadur Poudel;Rubin Thapa Magar;Adzemye Fovennso Bridget;Jae Kyung Sohng
    • Journal of Microbiology and Biotechnology
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    • v.33 no.7
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    • pp.949-954
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    • 2023
  • Type III polyketide synthase (PKS) found in bacteria is known as 1,3,6,8-tetrahydroxynaphthalene synthase (THNS). Microbial type III PKSs synthesize various compounds that possess crucial biological functions and significant pharmaceutical activities. Based on our sequence analysis, we have identified a putative type III polyketide synthase from Nocardia sp. CS682 was named as ThnA. The role of ThnA, in Nocardia sp. CS682 during the biosynthesis of 1,3,6,8 tetrahydroxynaphthalene(THN), which is the key intermediate of 1-(α-L-(2-O-methyl)-6-deoxymannopyranosyloxy)-3,6,8-trimethoxynaphthalene (IBR-3) was characterized. ThnA utilized five molecules of malonyl-CoA as a starter substrate to generate the polyketide 1,3,6,8-tetrahydroxynaphthalene, which could spontaneously be oxidized to the red flaviolin compound 2,5,7-trihydroxy-1,4-naphthoquinone. The amino acid sequence alignment of ThnA revealed similarities with a previously identified type III PKS and identified Cys138, Phe188, His270, and Asn303 as four highly conserved active site amino acid residues, as found in other known polyketide synthases. In this study, we report the heterologous expression of the type III polyketide synthase thnA in S. lividans TK24 and the identification of THN production in a mutant strain. We also compared the transcription level of thnA in S. lividans TK24 and S. lividans pIBR25-thnA and found that thnA was only transcribed in the mutant.

Mitochondrial Effects on the Physiological Characteristics of Lentinula edodes

  • Minseek Kim;Seong-Hyeok Yang;Hui-Gang Han;Eunbi Kim;Sinil Kim;Youn-Lee Oh;Hyeon-Su Ro
    • Mycobiology
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    • v.50 no.5
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    • pp.374-381
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    • 2022
  • In the mating of filamentous basidiomycetes, dikaryotic mycelia are generated through the reciprocal movement of nuclei to a monokaryotic cytoplasm where a nucleus of compatible mating type resides, resulting in the establishment of two different dikaryotic strains having the same nuclei but different mitochondria. To better understand the role of mitochondria in mushrooms, we created four sets of dikaryotic strains of Lentinula edodes, including B2×E13 (B2 side) and B2×E13 (E13 side), B5×E13 (B5 side) and B5×E13 (E13 side), E8×H3 (E8 side) and E8×H3 (H3 side), and K3×H3 (K3 side) and K3×H3 (H3 side). The karyotypes and mitochondrial types of the dikaryotic strains were successfully identified by the A mating type markers and the mitochondrial variable length tandem repeat markers, respectively. Comparative analyses of the dikaryotic strains on the mycelial growth, substrate browning, fruiting characteristics, and mitochondrial gene expression revealed that certain mitochondria are more effective in the mycelial growth and the production of fruiting body, possibly through the activated energy metabolism. Our findings indicate that mitochondria affect the physiology of dikaryotic strains having the same nuclear information and therefore a selection strategy aimed at mitochondrial function is needed in the development of new mushroom strain.

Experimental Study on the Triaxial Compressive Behaviour of Unsaturated Compacted Silt under Various Suction Levels (다양한 석션 레벨에서의 불포화실트의 삼축압축거동에 관한 실험적 연구)

  • Kim, Young-Seok;Oka, Fusao
    • Journal of the Korean Geotechnical Society
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    • v.24 no.1
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    • pp.25-35
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    • 2008
  • It has been recognized that the behaviour of unsaturated soil plays an important role in geomechanics. However, up to now, only a few experimental data are available for the technical difficulties related to both volume changes and suction measurements. In this study, the volume changes of unsaturated compacted silty soil were monitored with proximeter during various triaxial compression tests, which gave a realistic estimation in the volume changes of unsaturated soil sample. From the test results, the behaviours of wetting-induced collapses are observed during the drainage/water absorption tests. Under exhausted-drained conditions during shearing, the shear strength increases with an increasing initial suction. On the other hand, the volume changes become small with an increase in the initial suction. And, the volumetric strain during shearing is independent of the confining pressure.

Dynamic analysis of nanotube-based nanodevices for drug delivery in sports-induced varied conditions applying the modified theories

  • Shaopeng Song;Tao Zhang;Zhiewn Zhui
    • Steel and Composite Structures
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    • v.49 no.5
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    • pp.487-502
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    • 2023
  • In the realm of nanotechnology, the nonlocal strain gradient theory takes center stage as it scrutinizes the behavior of spinning cantilever nanobeams and nanotubes, pivotal components supporting various mechanical movements in sport structures. The dynamics of these structures have sparked debates within the scientific community, with some contending that nonlocal cantilever models fail to predict dynamic softening, while others propose that they can indeed exhibit stiffness softening characteristics. To address these disparities, this paper investigates the dynamic response of a nonlocal cantilever cylindrical beam under the influence of external discontinuous dynamic loads. The study employs four distinct models: the Euler-Bernoulli beam model, Timoshenko beam model, higher-order beam model, and a novel higher-order tube model. These models account for the effects of functionally graded materials (FGMs) in the radial tube direction, giving rise to nanotubes with varying properties. The Hamilton principle is employed to formulate the governing differential equations and precise boundary conditions. These equations are subsequently solved using the generalized differential quadrature element technique (GDQEM). This research not only advances our understanding of the dynamic behavior of nanotubes but also reveals the intriguing phenomena of both hardening and softening in the nonlocal parameter within cantilever nanostructures. Moreover, the findings hold promise for practical applications, including drug delivery, where the controlled vibrations of nanotubes can enhance the precision and efficiency of medication transport within the human body. By exploring the multifaceted characteristics of nanotubes, this study not only contributes to the design and manufacturing of rotating nanostructures but also offers insights into their potential role in revolutionizing drug delivery systems.