ENZYME AND MICROBIAL TECHNOLOGY
·
2023
·
IF 3.4
Shuangshuang Duan,Yonghua Chen,Guodong Wang,Zebin Li,Shitong Dong,Yingshuai Wu,Yuanwei Wang,Chunling Ma,Ruiming Wang
D -Allose is a rare cis-caprose with a wide range of physiological functions, which has a wide range of applications in medicine, food, and other industries. L -Rhamnose isomerase ( L -Rhi) is the earliest enzyme found to catalyze the production of D -allose from D -psicose. This catalyst has a high conversion rate, but its specificity for substrates is limited; thus, it cannot fulfill the requirements of industrial production of D -allose. In this study, L -Rhi derived from Bacillus subtilis was employed as the research subject, and D -psicose as the conversion substrate. Two mutant libraries were constructed through alanine scanning, saturation mutation, and rational design based on the analysis of the secondary structure , tertiary structure , and interactions with ligands of the enzyme. The yield of D -allose produced by these mutants was assessed; it was found that the conversion rate of mutant D325M to D -allose was increased by 55.73 %, and the D325S improved by 15.34 %, while mutant W184H increased by 10.37 % at 55 °C, respectively. According to modeling analysis, manganese (Mn 2+ ) had no significant effect on the production of D -psicose from D -psicose by L -Rhi. The results of molecular dynamics simulation demonstrated that the mutants W184H, D325M, and D325S had more stable protein structures while binding with the substrate D -psicose, as evidenced by its root mean square deviation (RMSD), root mean square fluctuation (RMSF), and binding free energy values. It was more conducive to binding D -psicose and facilitating its conversion to D -allose, providing the basis for the production of D -allose.
研究领域:
生物化学
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酶学
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生物技术
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结构生物学
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产品关联
DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2021
·
IF 4.098
Lingyuan Liu,Qianyi Zhang,Feng Li,Mei Wang,Jing Sun,Shuyun Zhu
Herein, fluorescent DNA-templated silver nanoclusters (DNA-AgNCs) with red emission were synthesized and utilized as novel probe to detect D-penicillamine (D-Pen) for the first time. D-Pen molecules contain a thiol which can combine with Ag to form a non-fluorescent ground state complex, inducing the aggregation of DNA-AgNCs followed by the fluorescence quenching. The quenching mechanism is well-studied and found to be a static quenching process. This method can detect D-Pen in the range of 0.025–0.7 μM with the detection limit as low as 8 nM, which is 1–3 orders of magnitude more sensitive than those based on other fluorescent nanoprobes. More importantly, the preparation procedure for DNA-AgNCs is fast and without the requirement of heavy metal ions. Thus, this detection strategy is time-saving and eco-friendly. Satisfactory recoveries have been acquired for monitoring D-Pen in human serum samples and pharmaceutical samples owing to the high sensitivity.
研究领域:
生物化学
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分析化学
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医学诊断
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产品关联
DOI
PubMed
高影响因子
Polymers
·
2022
·Vol. 14
·期 12
·2317
·
IF 4.967
Juliene Oliveira Campos de França,Deborah da Silva Valadares,Mateus Freitas Paiva,Sílvia Cláudia Loureiro Dias,José Alves Dias
Poly(lactic acid) (PLA) is an important polymer that is based on renewable biomass resources. Because of environmental issues, more renewable sources for polymers synthesis have been sought for industrial purposes. In this sense, cheaper monomers should be used to facilitate better utilization of less valuable chemicals and therefore granting more sustainable processes. Some points are raised about the need to study the total degradability of any PLA, which may require specific composting conditions (e.g., temperature, type of microorganism, adequate humidity and aerobic environment). Polymerization processes to produce PLA are presented with an emphasis on D,L-lactic acid (or rac-lactide) as the reactant monomer. The syntheses involving homogeneous and heterogeneous catalytic processes to produce poly(D,L-Lactic acid) (PDLLA) are also addressed. Additionally, the production of blends, copolymers, and composites with PDLLA are also presented exemplifying different preparation methods. Some general applications of these materials mostly dedicated to the biomedical area over the last 10–15 years will be pointed out.Keywords:poly(lactic acid)-PLA;polymerization based on D,L-lactic acid;D,L-lactide;stereo polymerization of D,L-lactic acid;blends of PLA;poly(DL-lactide-co-polymer);composites of PDLLA
研究领域:
高分子化学
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生物材料学
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环境科学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF BIOTECHNOLOGY
·
2018
·
IF 2.533
Binglin Li,Dandan Duan,Jiao Wang,Huanyu Li,Xiaoli Zhang,Binxia Zhao
Phospholipase D (PLD) was firstly hyperactivated by the bio-imprinting and, then, these hyperactivated PLD molecules were immobilized by adsorption and precipitation, followed by cross-linking. The high degree of conformational rigidification provided by intra- and intermolecular cross-linking can make PLD “remember” imprint-induced characteristics even in aqueous solutions. The obtained immobilized PLD showed the excellent catalytic performance. The maximum activity of immobilized PLD reached 166953 U/g protein , which was approximately 14 times higher than that of free form (11922 U/g protein ). Moreover, the selectivity of PLD was significantly enhanced after immobilization. The yield of PG and PA reached 94.0% and 5.96%, respectively. Compared with the serious hydrolysis in the free PLD (35.3% yield of PA), the side reaction was minimized in this work. This may be the first description of the remarkably high improvement of the activity and selectivity of PLD through the immobilization technology.
研究领域:
生物化学
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酶工程
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产品关联
DOI
PubMed
高影响因子
LANGMUIR
·
2020
·
IF 3.557
Mei Cui,Yuhao Duan,Yuanyuan Ma,Khaled W.A. Al-Shwafy,Yudong Liu,Xudong Zhao,Renliang Huang,Wei Qi,Zhimin He,Rongxin Su
Expansin has nonhydrolytic disruptive activity and synergistically acts with cellulases to enhance the hydrolysis of cellulose. The adsorption–desorption of expansin on noncellulosic lignin can greatly affect the action of expansin on lignocellulose. In this study, three lignins with different sources (kraft lignin (KL), sodium lignin sulfonate (SLS), and enzymatic hydrolysis lignin (EHL)) were selected as the substrates. The real-time adsorption–desorption of Bacillus subtilis expansin (BsEXLX1) on lignins was monitored using quartz crystal microgravimetry with dissipation (QCM-D). The effects of temperature and Tween 80 on the adsorption–desorption behaviors were also investigated. The results show that BsEXLX1 exhibited high binding ability on lignin and achieved maximum adsorption of 283.2, 273.8, and 266.9 ng cm–2 at 25 °C on KL, SLS, and EHL, respectively. The maximum adsorption decreased to 148.2–192.8 ng cm–2 when the temperature increased from 25 to 45 °C. Moreover, Tween 80 competitively bound to lignin and significantly prevented expansin adsorption. After irreversible adsorption of Tween 80, the maximum adsorption of BsEXLX1 greatly decreased to 33.3, 37.2, and 10.3 ng cm–2 at 25 °C on KL, SLS, and EHL, respectively. Finally, a kinetic model was developed to analyze the adsorption–desorption process of BsEXLX1. BsEXLX1 has a higher adsorption rate constant (kA) and a lower desorption rate constant (kD) on KL than on SLS and EHL. The findings of this study provide useful insights into the adsorption–desorption of expansin on lignin.
研究领域:
生物化学
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酶学
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材料科学
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产品关联
DOI
PubMed
高影响因子
Catalysts
·
2018
·Vol. 8
·期 10
·468
·
IF 3.465
Lu Zheng,Yining Sun,Jing Wang,He Huang,Xin Geng,Yi Tong,Zhi Wang
In this present study, we proposed a smart biomineralization method for creating hybrid organic–inorganic nanoflowers using a Co2+-dependent enzyme (D-psicose 3-epimerase; DPEase) as the organic component and cobalt phosphate as the inorganic component. The prepared nanoflowers have many separated petals that have a nanometer size. Under optimum conditions (60 °C and pH of 8.5), the nanoflower can display its maximum activity (36.2 U/mg), which is about 7.2-fold higher than free DPEase. Furthermore, the immobilized DPEase presents enhanced pH and thermal stabilities. The DPEase-nanoflower maintained about 90% of its activity after six reaction cycles, highlighting its excellent reusability.Keywords:nanoflower;immobilization;metal-dependent enzyme;D-psicose;DPEase
研究领域:
生物化学
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酶工程
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DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2025
·
IF 6.2
Yuxin Wang,Zijian Tan,Hongli Wei,Nan Zhang,Lifei Wang,Yifan Yang,Weidong Liu,Leilei Zhu
D-Tagatose, a promising sugar substitute with various functional properties and commercial applications, can be enzymatically converted from D-fructose by tagatose 4-epimerase. The development of an efficient tagatose 4-epimerase that catalyzes the conversion of D-fructose into D-tagatose is essential to make the production technology of D-tagatose applicable. In this study, tagatose 4-epimerase from Thermotogae (TsT4Ease) was engineered through semi-rational design and directed evolution, resulting in a 2.8-fold improvement in catalytic activity compared to the wild type (WT). The production of D-tagatose reached 42 g/L in 2 h. Crystal structure analysis determined the structural features with a common (α/β)8-TIM barrel and a Zn2+-binding architecture at the active center. Subsequent molecular dynamics (MD) simulations revealed that the substitutions improved substrate binding energy and stabilized the active pocket. This study offers new insights into the structure–function relationship of TsT4Ease and provides a candidate tagatose 4-epimerase.
研究领域:
生物化学
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蛋白质工程
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结构生物学
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DOI
PubMed
高影响因子
Cell Death & Disease
·
2025
·Vol. 16
·期 1
·1-11
·
IF 9.6
Chu Xiufeng,Zhang Ting,Bukhari Ihtisham,Hu Mei,Xu Jixuan,Xing Yamin,Liang Xinfeng,Zhang Zisen,Zheng Pengyuan
Gasdermin D (GSDMD) is a critical pyroptosis mediator, consisting of one N-terminal pore-forming domain and one C-terminal auto-inhibitory domain. The free N-terminal domain (GD-NT), which is released through caspase-1/11 cleavage, exhibits distinct features from the full-length GSDMD (GD-FL), including oligomerization, membrane translocation, and pore-formation. However, the underlying mechanisms are not well elucidated. Here, we found that GD-NT, but not GD-FL, was massively ubiquitinated in cells. The K63-linked polyubiquitination of GD-NT at Lys236/237 (human/mouse), catalyzed by TRAF1, directly prompted its membrane translocation and pore-formation during pyroptosis. Inhibition of GD-NT ubiquitination via site-directed mutations or the UBA1 inhibitor PYR-41 suppressed cell death in several pyroptosis cell models. Additionally, applying PYR-41 in septic mice efficiently suppressed the release of IL-18 and TNFα. Thus, GD-NT ubiquitination is a key regulatory mechanism controlling its membrane localization and activation, which may provide a novel target for modulating immune activity in pyroptosis-related diseases.
研究领域:
免疫学
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细胞生物学
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产品关联
DOI
PubMed
高影响因子
Catalysts
·
2020
·Vol. 10
·期 9
·1057
·
IF 3.52
Jing Wang,Sheng Xu,Yang Pang,Xin Wang,Kequan Chen,Pingkai Ouyang
To achieve efficient bio-production of phospholipase D (PLD), PLDs from different organisms were expressed inE.coli. An efficient secretory expression system was thereby developed for PLD. First, PLDs fromStreptomycesPMF andStreptomyces racemochromogeneswere separately over-expressed inE.colito compare their transphosphatidylation activity based on the synthesis of phosphatidylserine (PS), and PLDPMFwas determined to have higher activity. To further improve PLDPMFsynthesis, a secretory expression system suitable for PLDPMFwas constructed and optimized with different signal peptides. The highest secretory efficiency was observed when the PLD * (PLDPMFwith the native signal peptide Nat removed) was expressed fused with the fusion signal peptide PelB-Nat inE. coli. The fermentation conditions were also investigated to increase the production of recombinant PLD and 10.5 U/mL PLD was ultimately obtained under the optimized conditions. For the application of recombinant PLD to PS synthesis, the PLD properties were characterized and 30.2 g/L of PS was produced after 24 h of bioconversion when 50 g/L phosphatidylcholine (PC) was added.Keywords:phospholipase D;phosphatidylserine;secretory expression;enzymatic catalysis
研究领域:
生物化学
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分子生物学
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产品关联
DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2023
·
IF 8.2
Jing-Jing Liu,Zhan-Yi Wang,Bin-Bin Jiang,Shu-Qin Gao,Ying-Wu Lin
Nonenzymatic glycation and the subsequent accumulation of advanced glycation end-products (AGEs) in proteins are factors underlying long-term pathogenesis in diabetes. The study of protein glycation is crucial for elucidating their relationship with diabetes mellitus and related disorders. This study explores the interaction between d -ribose and human myoglobin (HMb), as well as the protective effect of thymoquinone (TQ) on glycation. A time-dependent in-vitro glycation study was performed to investigate the mechanism of d -ribose-induced structural interference of HMb in the absence and presence of TQ. Spectroscopic and proteomic analysis indicated that the presence of TQ significantly reduced the total amount of AGEs while maintaining structural characteristics of HMb. 14 glycated sites on HMb were further identified via liquid chromatography-tandem mass spectrometry (LC-MS/MS) after incubation with d -ribose for 12 h, predominantly interacting with lysine residues. TQ was found to disrupt this interaction, reducing the glycated sites from 14 to 12 sites and the percentage of glycated peptides from 26.50 % to 12.97 %. Additionally, there was a significant decrease in the degree of glycation at the same sites. In summary, our findings suggest that TQ has the potential to act as an anti-glycation agent and provide a comprehensive understanding underlying the inhibition mechanism of glycation.
研究领域:
生物化学
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医学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Huayang Tang,Yian Chen,Dexun Fan,Fengguang Zhao,Shuangyan Han
D-allulose, a low-calorie rare sugar catalyzed by D-allulose 3-epimerase (DAE), is highly sought after for its potential health benefits. However, poor reusability and stability of DAE limited its popularization in industrial applications. Although metal-organic frameworks (MOFs) offer a promising enzyme platform for enzyme immobilization, developing customized strategies for MOF immobilization of enzymes remains challenging. In this study, we introduce a designable strategy involving the construction of bimetal-organic frameworks (ZnCo-MOF) based on metal ions compatibility. The DAE@MOFs materials were prepared and characterized, and the immobilization of DAE and the enzymatic characteristics of the MOF-immobilized DAE were subsequently evaluated. Remarkably, DAE@ZnCo-MOF exhibited superior recyclability which could maintain 95 % relative activity after 8 consecutive cycles. The storage stability is significantly improved compared to the free form, with a relative activity of 116 % remaining after 30 days. Molecular docking was also employed to investigate the interaction between DAE and the components of MOFs synthesis. The results demonstrate that the DAE@ZnCo-MOF exhibited enhanced catalytic efficiency and increased stability. This study introduces a viable and adaptable MOF-based immobilization strategy for enzymes, which holds the potential to expand the implementation of enzyme biocatalysts in a multitude of disciplines.
研究领域:
生物化学
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食品科学
·
材料科学
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产品关联
DOI
PubMed
高影响因子
FLUID PHASE EQUILIBRIA
·
2015
·
IF 2.2
Liying Cai,Tingping Xie,Suying Zhao,Huidong Zheng,Dan Wu
The solubility of d -camphor-10-sulfonic acid (D-CSA, CAS No. 3144-16-9) in seven solvents (ethyl acetate, methyl acetate , n -butyl acetate, isopropyl acetate, glacial acetic acid, acetic anhydride and propionic acid) was measured under atmospheric pressure. The solubility data were correlated by the modified Apelblat equation, the λh equation, the Wilson model and the UNIQUAC model, respectively. And the thermodynamic parameters of D-CSA, molar dissolution enthalpy and entropy, were determined by the Van’t Hoff equation to explain its dissolving behavior in each solvent.
研究领域:
化学工程
·
物理化学
✓ 已验证
DOI
高影响因子
危险品化学品经营许可证（带存储）