INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2019
·
IF 4.784
Meng-Qiu Xu,Fei-Long Li,Wen-Qian Yu,Rui-Fang Li,Ye-Wang Zhang
Cofactor regeneration is an important method to avoid the consumption of large quantities of oxidized cofactor NAD + in enzyme-catalyzed reactions. Herein, glycerol dehydrogenase (GDH) and NADH oxidase preparations by aggregating enzymes with ammonium sulphate followed by cross-linking formed aggregates for effective regeneration of NAD + . After optimization, the activity of combi-CLEAs and separate CLEAs mixtures were 950 and 580 U/g, respectively. And the catalytic stability of combi-CLEAs against pH and temperature was superior to the free enzyme mixture. After ten cycles of reuse, the catalytic efficiency could still retain 63.3% of its initial activity, indicating that the constructed combi-CLEAs system had excellent reusability. Also, the conversion of glycerol to 1,3-dihydroxyacetone (DHA) was improved by the constructed NAD + regeneration system, resulting in 4.6%, which was 2.5 times of the free enzyme system. Thus, wide applications of this co-immobilization method in the production of various chiral chemicals could be expected in the industry for its high efficiency at a low cost.
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
BMC Veterinary Research
·
2022
·Vol. 18
·期 1
·1-14
·
IF 2.792
Hao Fei,Xie Xing,Feng Zhixin,Chen Rong,Wei Yanna,Liu Jin,Xiong Qiyan,Shao Guoqing,Lin Johnson
Background Mycoplasma hyopneumoniae (M. hyopneumoniae) is the etiological agent of enzootic pneumonia, a highly infectious swine respiratory disease that distributed worldwide. The pathogenesis and virulence factors of M. hyopneumoniae are not fully clarified. As an important virulence factor of bacteria, nicotinamide adenine dinucleotide (NADH) oxidase (NOX) participates in host-pathogen interaction, however, the function of NOX involved in the pathogenesis of M. hyopneumoniae is not clear.Results In this study, significant differences in NOX transcription expression levels among different strains of M. hyopneumoniae differed in virulence were identified, suggesting that NOX may be correlated with M. hyopneumoniae virulence. The nox gene of M. hyopneumoniae was cloned and expressed in Escherichia coli, and polyclonal antibodies against recombinant NOX (rNOX) were prepared. We confirmed the enzymatic activity of rNOX based on its capacity to oxidize NADH to NAD+. Flow cytometry analysis demonstrated the surface localization of NOX, and subcellular localization analysis further demonstrated that NOX exists in both the cytoplasm and cell membrane. rNOX was depicted to mediate adhesion to immortalized porcine bronchial epithelial cells (hTERT-PBECs). Pre-neutralizing M. hyopneumoniae with anti-rNOX antibody resulted in a more than 55% reduction in the adhesion rate of high- and low-virulence M. hyopneumoniae strains to hTERT-PBECs. Moreover, a significant difference appeared in the decline in CCU50 titer between virulent (168) and virulence-attenuated (168L) strains. NOX not only recognized and interacted with host fibronectin but also induced cellular oxidative stress and apoptosis in hTERT-PBECs. The release of lactate dehydrogenase by NOX in hTERT-PBECs was positively correlated with the virulence of M. hyopneumoniae strains.Conclusion sNOX is considered to be a potential virulence factor of M. hyopneumoniae and may play a significant role in mediating its pathogenesis.
研究领域:
微生物学
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兽医学
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分子生物学
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病理学
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产品关联
DOI
PubMed
高影响因子
ACS Catalysis
·
2018
·
IF 11.384
Yizhou Wu,Jesse Ward-Bond,Donglin Li,Shaohua Zhang,Jiafu Shi,Zhongyi Jiang
Graphitic carbon nitride (g-C3N4) is an emergent metal-free photocatalyst because of its band position, natural abundance, and facile preparation. Synergetic intensification of charge generation and charge transfer of g-C3N4 to increase solar-to-chemical efficiency remains a hot yet challenging issue. Herein, a nanoshell with two moieties of α-Fe2O3 and carbon (C) is in situ formed on the surface of a g-C3N4 core through calcination of Fe3+/polyphenol-coated melamine, thus acquiring g-C3N4@α-Fe2O3/C core@shell photocatalysts. The α-Fe2O3 moiety acts as an additional photosensitizer, offering more photogenerated electrons, whereas the C moiety bridges a “highway” to facilitate the electron transfer either from α-Fe2O3 moiety to g-C3N4 or from g-C3N4 to C moiety. By tuning the proportion of these two moieties in the nanoshell, a photocurrent density of 3.26 times higher than pristine g-C3N4 is obtained. When utilized for photocatalytic regeneration of reduced nicotinamide adenine dinucleotide (NADH, a dominant cofactor in biohydrogenation reaction), g-C3N4@α-Fe2O3/C exhibits an equilibrium NADH yield of 76.3% with an initial reaction rate (r) of 7.7 mmol h–1 g–1, among the highest r for photocatalytic NADH regeneration ever reported. Manipulating the coupling between charge generation and charge transfer may offer a facile, generic strategy to improve the catalytic efficiency of a broad range of photocatalysts other than g-C3N4.
研究领域:
材料科学
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光化学
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催化
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纳米技术
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产品关联
DOI
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2025
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IF 4.6
Xinyu Lan,Guojun Weng,Jianjun Li,Jian Zhu,Junwu Zhao
Surface metal deposition is an important method for tuning the optical property of plasmonic nanoparticles. In this work, we tried to deposit Cu shell on the surface of gold nanorods (Au NRs) by dihydronicotinamide adenine dinucleotide (NADH)-mediated reaction. Cu 2+ were reduced to Cu 0 through the gold-catalyzed oxidization of NADH. The deposition of Cu shell onto the Au NRs surface resulted in the obvious red-shift of longitudinal localized surface plasmon resonance peak. And the red-shift of longitudinal peak increased with the concentrations of NADH and Cu 2+ . The TEM images showed that a transparent layer structure with a different contrast around Au NRs was clearly displayed. And this transparent layer structure with the low contrast was generally considered to be pure Cu shell. However, the results from energy dispersive spectrometer (EDS) and X-ray photoelectron spectroscopy (XPS) indicated that the transparent layer was not just pure Cu composition, but contained NADH/NAD + and Cu 2+ components. This grown complex dielectric shell resulted in the reduced surface enhanced Raman scattering (SERS) activity of Au NRs. Based on the decreased SERS intensity, we demonstrated a SERS method for Cu 2+ detection using R6G as Raman reporter. The decreased SERS intensity had a linear correlation with the concentration of Cu 2+ in the range of 10 −6 –10 −2 M, with a detection limit of 3.89 × 10 −6 M. This study provides new insights about Cu deposition on Au NRs, which will further promote the spectral sensing applications with gold-catalyzed Cu 2+ reduction on plasmonic nanoparticles by NADH.
研究领域:
材料科学
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分析化学
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生物医学工程
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产品关联
DOI
PubMed
高影响因子
PHYSICAL CHEMISTRY CHEMICAL PHYSICS
·
2020
·Vol. 22
·期 39
·22746-22757
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IF 3.43
Wenting Wei,Jiaxuan Li,Huiqin Yao,Keren Shi,Hongyun Liu
Herein, a new type of lanthanide coordination polymer film made up of europium (Eu(III)) and poly(N-methacryloylglycine) (Eu(III)–PMAG) was prepared on an ITO electrode surface driven by the coordination between N-methacryloylglycine (MAG) and Eu(III) through a single-step polymerization process. The fluorescence signal of Eu(III)–PMAG films at 617 nm originating from Eu(III) could be well retained in the buffer solution but was regulated by the concentration of Cu(II) and the complexing agent EDTA. The switching of fluorescence by Cu(II) was attributed to the inhibition of the “antenna effect” between Eu(III) and the MAG ligand in the films. The coexistence of reduced β-nicotinamide adenine dinucleotide (NADH) in the solution can apparently quench the fluorescence of Eu(III)–PMAG films through the internal filtration effect of UV absorbance overlapping the excitation wavelength, but itself exhibiting a fluorescence emission at 468 nm. In addition, the electrocatalytic oxidation of NADH with the help of the ferrocenedicarboxylic acid (FcDA) probe demonstrated a cyclic voltammetry (CV) signal at 0.45 V (vs. SCE). Based on various reversible stimulus-responsive behaviours, a 4-input/10-output logic network was built using Cu(II), EDTA, NADH and FcDA as inputs and the signals of fluorescence from Eu(III)–PMAG (617 nm) and NADH (468 nm), the CV response from FcDA and the UV-vis absorbance from the Cu(II)–EDTA complex as outputs. Meanwhile, 6 different functional logic devices were constructed based on the same versatile platform, including a 2-to-1 encoder, a 1-to-2 decoder, a 1-to-2 demultiplexer, a parity checker, a transfer gate and a reprogrammable 3-input/2-output keypad lock. Combined with the new type of lanthanide coordination polymer film, NADH played central roles in designing sophisticated computing systems with its fluorescence, UV and electrocatalytic properties. This work might provide a novel avenue to develop intelligent multi-analyte sensing and information processing at the molecular level based on one single platform.
研究领域:
生物化学
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材料科学
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分子生物学
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DOI
PubMed
高影响因子
3 Biotech
·
2024
·Vol. 15
·期 1
·1-8
·
IF 2.6
Feng Xinming,Cui Xinyu,Wang Kun,Liu Juanjuan,Meng Dongdong
6-Phosphogluconate dehydrogenases (6PGDHs) are widely existing as reduced cofactor (NADH/NADPH) regeneration biocatalysts. Herein, a thermostable 6PGDH from Hydrogenobacter thermophilus (Ht6PGDH) was overexpressed in Escherichia coli and enzymologically characterized. Ht6PGDH exhibited exceptional stability and catalytic activity under high-temperature conditions, with an optimum temperature of 85 °C and the ability to maintain high activity for prolonged periods at 70 °C, which could be purified through a one-step heat treatment. Moreover, Ht6PGDH exhibited a preference for NAD + with a K m value of 0.4 mM and a k cat value of 28.6 s⁻ 1 , demonstrating a significant preference over NADP + . These properties render Ht6PGDH a potentially valuable enzyme for high-temperature bioconversion and in vitro synthetic biosystems. Additional research showed that Ht6PGDH excelled in the regeneration of NADH, achieving efficient lactate production when integrated into an in vitro synthetic biosystem containing lactate dehydrogenase (LDH). Furthermore, the cascade reaction of Ht6PGDH with glucose-6-phosphate dehydrogenase (G6PDH) was explored for NADH regeneration using starch as the substrate, further validating its potential application in complex biosynthetic systems.
研究领域:
生物化学
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酶工程
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合成生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
New Biotechnology
·
2021
·
IF 5.079
Wen-Bin Su,Fei-Long Li,Xue-Yong Li,Xiao-Man Fan,Rui-Jiang Liu,Ye-Wang Zhang
L-Tagatose, a promising building block in the production of many value-added chemicals, is generally produced by chemical routes with a low yield, which may not meet the increasing demands. Synthesis of l -tagatose by enzymatic oxidation of d -galactitol has not been applied on an industrial scale because of the high cofactor costs and the lack of efficient cofactor regeneration methods. In this work, an efficient and environmentally friendly enzymatic method containing a galactitol dehydrogenase for d -galactitol oxidation and a water-forming NADH oxidase for regeneration of NAD + was first designed and used for l -tagatose production. Supplied with only 3 mM NAD + , subsequent reaction optimization facilitated the efficient transformation of 100 mM of d -galactitol into l -tagatose with a yield of 90.2 % after 12 h (obtained productivity: 7.61 mM.h −1 ). Compared with the current chemical and biocatalytic methods, the strategy developed avoids by-product formation and achieves the highest yield of l -tagatose with low costs. It is expected to become a cleaner and more promising route for industrial biosynthesis of l -tagatose.
研究领域:
生物技术
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生物化学
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酶学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF COLLOID AND INTERFACE SCIENCE
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2023
·
IF 9.9
Chun-Hui Gao,Shi-Ming Zhang,Fang-Fang Feng,San-San Hu,Qian-Fan Zhao,Yong-Zheng Chen
Visible-light-driven nicotinamide adenine dinucleotide (NADH) regeneration is one of the most effective measures, and cadmium sulfide (CdS) materials are typically used as low-cost photocatalysts. The CdS photocatalysts, however, still suffer from low regeneration efficiency and poor cycle stability. In this work, the CdS quantum dots (QDs) less than 10 nm embedded onto silica gel (CdS QDs/Silica gel) were constructed for visible-light-driven NADH regeneration by a successive ionic layer adsorption reaction and ball milling method. Results demonstrate that the photosensitivity of the CdS QDs/Silica gel composite was 31 times higher than that of the bulk CdS. Moreover, the conduction band (CB) edge of the CdS QDs/Silica gel composite is −1.34 eV, which is more negative 0.5 eV than that of the bulk CdS. The obtained CdS QDs/Silica gel composites showed the highest NADH regeneration yields of 68.8% under visible-light (LED, 420 nm) illumination and can be reused for over 40 cycles. Finally, the bioactivity of NADH toward enzyme catalysis is further confirmed by the hydrogenation of benzaldehyde to benzyl alcohol catalyzed with an alcohol dehydrogenase as enzyme catalysis.
研究领域:
生物化学
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材料科学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2019
·
IF 6.35
Zongwen Wang,Qian Chen,Yingying Zhong,Xinhui Yu,Yongning Wu,FengFu Fu
Many studies have demonstrated that the extracellular domain of human epidermal growth factor receptor 2 (HER2 ECD) level in serum can act as a breast cancer biomarker and serve as a monitoring neoadjuvant therapy of breast cancer. In this study, we developed a sensitive ascorbic acid (AA)-mediated AuNBPs (gold nanobipyramids) growth method with NADH (reduced nicotinamide adenine dinucleotide I) assistance, and we further fabricated a high-resolution multicolor immunosensor for sensitive visual detection of HER2 ECD in serum by using AuNBPs as signal and antibody as recognition probe. The NADH-assisted AA-mediated method effectively suppressed color formation in the blank and greatly improved the sensitivity of mediating AuNBPs growth, allowing us to use a low concentration of AA to mediate AuNBPs growth to generate more colorful and clearer color changes. The proposed multicolor immunosensor has higher resolution and more color changes corresponding to HER2 ECD concentrations. It can be used to detect as low as 0.5 ng/mL of HER2 ECD by bare eye observation and 0.05 ng/mL of HER2 ECD by UV–visible spectrophotometry. Using the immunosensor, we have successfully detected HER2 ECD in human serum with a recovery of 94%–96% and an RSD (n = 5) < 5%. The results obtained with our immunosensor were consistent with those obtained with ELISA, verifying the immunosensor has good accuracy. The immunosensor exhibited a vivid multicolor change, has low visual detection limit, excellent specificity and reproducibility, and robust resistance to matrix. All the above features makes our immunosensor a promising assay for the early diagnosis of HER2-dependent breast cancers in clinical diagnosis.
研究领域:
生物化学
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医学诊断
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纳米技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 10
·1-9
·
IF 6.232
Yang Hongxiao,Hou Jiagang,Wang Zhaohui,Zhou Qiuxia,Xu Caixia
A superior NADH sensing platform was constructed based on porous PtAg nanoshells supported on reduced graphene oxide (PtAg/rGO) in the absence of any enzymes and redox mediators. The PtAg/rGO composite was prepared via one-step reduction combined with galvanic replacement reaction. The as-made PtAg/rGO assembles multiple structural advantages of coherent conductive matrix, rich electroactive sites, and high specific surface area, accompanied by the unique alloying effect. The PtAg/rGO possesses adequate active reaction sites and fluent electron transport pathway towards the electrocatalytic NADH oxidation, thus presenting significantly increased oxidation current and negative shift of 330 mV in applied potential relative to the bare GCE. By virtues of the outstanding electrocatalytic activity, PtAg/rGO exhibits effective amperometric detection of NADH at 0.15 V within a wide linear concentration range of 2–2378 μM, a high sensitivity of 92.62 μA mM −1 cm −2 , low detection limit of 0.2 μM, and long-term detection over 2500 s. Moreover, the as-constructed biosensors can achieve accurate NADH detection in human serum samples, indicating its promising application feasibility in fundamental and clinic research.
Porous PtAg alloy nanoshells supported on reduced graphene oxide (PtAg/rGO) was prepared via a facile one-step reduction and spontaneous replacement reaction strategy. A sensitive and highly stable electrochemical biosensor based on PtAg/rGO is constructed for the quantitative detection of NADH at low applied potential.
研究领域:
电化学
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生物传感
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纳米材料
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY
·
2018
·Vol. 93
·期 8
·2351-2358
·
IF 2.587
Meng-Yao Zhuang,Xiao-Ping Jiang,Xiao-Min Ling,Meng-Qiu Xu,Yi-Hao Zhu,Ye-Wang Zhang
BACKGROUND Nicotinamide cofactor-dependent oxidoreductases have been widely applied to the bioproduction of varieties of useful compounds. Efficient cofactor regeneration is often required for these oxidoreductase-catalyzed reactions. Herein, enzymatic production of 1, 3-dihydroxyacetone (DHA) via immobilized enzymes with in situ cofactor regeneration was reported. RESULTS Glycerol dehydrogenase (GDH) and NADH oxidase (Nox) were immobilized on two carriers: macroporous resin and epoxy functionalized magnetic nanoparticles (EFMN). Higher enzyme loading capacity and activity recovery were achieved on EFMN. A high immobilization yield of 90% and >70% recovery activity were obtained for both enzymes using EFMN as carriers. Besides, both immobilized enzymes showed enhanced pH stability and temperature tolerance. Co-immobilized and mixed immobilized enzymes were evaluated for synthesis of DHA, and mixed immobilized enzymes showed higher catalytic rate. The mixed reaction system also was investigated in view of various operational factors. Under the optimal conditions, a high DHA concentration with mixed immobilized enzymes system can reach up to 3.5 mmol L –1 , which was about four times higher than that without cofactor regeneration system. CONCLUSION The immobilized GDH and Nox on magnetic nanoparticles showed enhanced stabilities. A cofactor regeneration system with immobilized enzymes could be utilized for efficient production of DHA from inexpensive substrates. © 2018 Society of Chemical Industry
研究领域:
生物化学
·
酶工程
✓ 已验证
产品关联
DOI
高影响因子
ACS Catalysis
·
2023
·
IF 12.9
Song Wang,Xiewen Wu,Jing Fang,Feng Zhang,Yanli Liu,Hongbo Liu,Yu He,Min Luo,Run Li
The design of a highly efficient polymer photocatalyst is an essential prerequisite for photo-biocatalysis in a polymer–enzyme coupled system, which offers an important platform for the synthesis of pharmaceuticals and fine chemicals. However, current polymers still suffer drawbacks of poor water wettability, low visible light absorption, and high recombination rate of photoinduced electron/hole pairs, which severely limit their application in photo-biocatalysis. Herein, we demonstrate a polymer/polymer double-shell hollow nanostructure (PCN@PDBTS-HN) via a facile two-step polymerization with polymeric carbon nitride (PCN) and poly-dibenzothiophene sulfone (PDBTS) as internal and external shells, respectively. The covalent connection of those two polymer layers ensures the spontaneous electron transfer from PDBTS to PCN, as reflected in X-ray photoelectron spectroscopy (XPS), electron paramagnetic resonance (EPR), and fluorescence spectra, which could effectively accelerate the charge separation and migration. Thanks to its improved light-harvesting, increased hydrophilicity, and enhanced redox capacity, the obtained PCN@PDBTS-HN demonstrates an excellent catalytic efficiency in visible-light-promoted NADH regeneration with a conversion of 85.4% being achieved in 40 min (turnover frequency of 0.859 mmol g–1 h–1). This value is 1.4 and 2.6 times higher than that of pristine PCN and PDBTS, respectively, indicating an essential role of Z-scheme heterojunction in improving the catalytic efficiency. In addition, PCN@PDBTS-HN also displays excellent photocatalytic selectivity for 1,4-NADH regeneration, remarkable photostability, and good biocompatibility. A total amount of 2.12 mM methanol and 8.39 mM l-glutamate was accumulated in the polymer/alcohol dehydrogenase (YADH) and polymer/glutamate dehydrogenase (GDH) coupled system, respectively, implying high flexibility of fine chemical production via the photo-biocatalytic approach.
研究领域:
生物化学
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材料科学
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光催化
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）