iScience
·
2025
·
IF 4.1
Shuai Liu,Yi Li,Lin Quan,Hai-Xia Liu,Yang Luo,Yong-Zhong Wang
SUMMARY Optimizing cellulase biosynthesis in B. subtilis is crucial for enhancing enzymatic yield in lignocellulosic biomass conversion. However, the regulatory mechanisms linking intracellular NAD(H/ + ) levels to cellulase production remain elusive. In this study, we systematically screened thirteen genes associated with NAD + biosynthesis and NADH regeneration in B. subtilis Z2. Employing a modular engineering strategy with four distinct modules, we directed metabolic flux to enhance NAD + biosynthesis and NADH regeneration. Key genes ( ycel , nadV , nadM , mdh , and sucB ) were identified, and their co-expression in B. subtilis Z2 resulted in a 13.09-fold increase in intracellular NADH levels and a consequential 2.24- and 2.04-fold enhancement in the filter paper-hydrolyzing (FPase [representing total cellullase]) activity and carboxymethylcellulase (CMCase [representing endoglucanase]) activity, respectively. Experimental validations, including antagonist LaCl 3 treatment and spcF gene deletion, unequivocally established the calcium signaling pathway's role in regulating cellulase gene overexpression in response to elevated intracellular NAD(H/ + ) levels.
研究领域:
微生物学
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生物化学
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合成生物学
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DOI
PubMed
高影响因子
ANALYST
·
2018
·Vol. 143
·期 21
·5278-5284
·
IF 3.864
Meng Wang,Xianwen Kan
In this study, an exfoliated graphite paper (EGP) was employed as a supported electrode for prussian blue (PB) and gold nanoparticles (AuNPs) modification to fabricate a novel electrochemical sensor. The scanning electron microscopy image showed the multilayer structure of the sensor with PB and AuNPs uniform coating. Due to the high surface area and good conductivity of the multilayer structure, as well as the conductivity of AuNPs, the prepared sensor exhibited an obvious improvement for sensitive detection of nicotinamide adenine dinucleotide (NADH) with a linear range of 5.0 × 10−7 mol L−1–1.0 × 10−3 mol L−1. Moreover, the modified PB film endowed the sensor with good electrocatalytic activity for hydrogen peroxide (H2O2) sensing. Under the optimal conditions, the linear range and limit of detection for H2O2 sensitive detection were 5.0 × 10−8 mol L−1–1.0 × 10−3 mol L−1 and 4.8 × 10−9 mol L−1, respectively. The sensor can also be used for detection in real samples with satisfactory results. The advantages of low-cost, easy modification, multilayer structure, and good conductivity made the EGP an alternative candidate for supported electrode materials in the preparation and application of various electrochemical sensors.
研究领域:
分析化学
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生物传感器
·
电化学
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DOI
PubMed
高影响因子
LANGMUIR
·
2013
·
IF 4.187
Panpan Li,Xu Chen,Wensheng Yang
Controlling the macroscopic organization of self-assembled peptide nanostructures on a solid surface is a key challenge in enabling their technological applications. Here, we report a simple approach to achieve the horizontally organized self-assembly of dipeptides by introducing graphene sheets. We show at the first time the formation of a macroscopic-scale, high-density, and ordered interlaced array of peptide nanowires and graphene composite (PNWs-G) on a silicon surface under mild conditions. The action of graphene sheets in the formation of the organized bionanostructure was preliminarily investigated. Furthermore, due to the introduction of graphene, the electronic conductivity of the bionanostructures was greatly improved, which is very beneficial for their applications in bioelectrochemical and nanoelectronic devices. As an applied example, the significantly enhanced electrochemical sensing performance for dihydronicotinamide adenine dinucleotide (NADH) was also demonstrated at the PNWs-G modified electrode relative to the alone component and unordered composite modified electrodes. The simple and mild approach described here opens a new avenue for the fabrication of macroscopic-scale organized self-assembled peptide bionanostructures on a solid surface, which should be capable of being extended to other biosystems based on graphite surface-template assembly, allowing a variety of functional bionanostructures to be fabricated and used in practical applications.
研究领域:
生物纳米技术
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材料科学
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电化学
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DOI
PubMed
高影响因子
CHEMICAL PHYSICS LETTERS
·
2019
·
IF 1.901
Simin Cao,Zhongneng Zhou,Haoyang Li,Menghui Jia,Yangyi Liu,Mengyu Wang,Mengjie Zhang,Sanjun Zhang,Jinquan Chen,Jianhua Xu,Jay R. Knutson
The metabolic cofactor and energy carrier NADH (nicotinamide adenine dinucleotide, reduced) has fluorescence yield and lifetime that depends strongly on conformation, a fact that has enabled metabolic monitoring of cells via FLIM (Fluorescence Lifetime Microscopy). Using femtosecond fluorescence upconversion, we show that this molecule in solution participates in ultrafast self-quenching along with both bulk solvent relaxation and spectral relaxation on 1.4 and 26 ps timescales. This, in effect, means up to a third of NADH is effectively “dark” for FLIM in the 400–500 nm observation window commonly employed. Methods to compensate for, avoid or measure dark species corrections are outlined.
研究领域:
生物物理学
·
细胞生物学
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代谢组学
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DOI
PubMed
高影响因子
BIOELECTROCHEMISTRY
·
2019
·
IF 4.474
Xueyan Li,Xianwen Kan
In this work, a novel electrochemical sensing platform was designed and fabricated by the modification of boronic acid functionalized carbon nanodots (B-CNDs) and poly(thionine) (pTHI) on an electrode surface. B-CNDs can not only accelerate electron transfer but also covalently interact with cis-diol groups of dihydronicotinamide adenine dinucleotide (NADH) through functionalized boronic acid groups. Meanwhile, pTHI served as an inner reference element to provide a built-in correction, which enabled the sensor to detect NADH with high accuracy and reliability based on a ratiometric signal (∆I NADH /∆I THI ). The electrochemical experimental results demonstrated that the ratiometric strategy-based sensor possessed good selectivity and high sensitivity. A linear range of 5.0 × 10 −7 - 2.0 × 10 −4 mol/L for NADH detection was obtained with a limit of detection of 1.5 × 10 −7 mol/L. The sensor has been applied to analyze NADH in human serum samples with satisfactory results. The simple and effective ratiometric strategy reported here can be further used to prepare electrochemical sensors for selective, sensitive, and reliable detection of other cis-diol compounds.
研究领域:
生物化学
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分析化学
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DOI
PubMed
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2016
·Vol. 100
·期 12
·5607-5618
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IF 3.376
Li Mu,Su Yinglong,Chen Yinguang,Wan Rui,Zheng Xiong,Liu Kun
The heterotrophic denitrification requires the participation of electrons which are derived from direct electron donor (usually nicotinamide adenine dinucleotide (NADH)), and the electrons are transferred via electron transport system in denitrifiers and then consumed by denitrifying enzymes. Despite the reported electron transfer ability of humic substances (HS), the influences of fulvic acid (FA), an ubiquitous major component of HS, on promoting NADH generation, electron transfer, and consumption in denitrification process have never been reported. The presence of FA, compared with the control, was found not only significantly improved the total nitrogen (TN) removal efficiency (99.9 % versus 74.8 %) but remarkably reduced the nitrite accumulation (0.2 against 43.8 mg/L) and N 2 O emission (0.003 against 0.240 mg nitrogen/mg TN removed). The mechanisms study showed that FA increased the metabolism of carbon source via glycolysis and tricarboxylic acid (TCA) cycle pathways to produce more available NADH. FA also facilitated the electron transfer activities from NADH to denitrifying enzymes via complex I and complex III in electron transport system, which improved the reduction of nitrate and accelerated the transformations of nitrite and N 2 O, and lower nitrite and N 2 O accumulations were therefore observed. In addition, the consumption of electrons in denitrification was enhanced due to FA stimulating the synthesis and the catalytic activity of key denitrifying enzymes, especially nitrite reductase and N 2 O reductase. It will provide an important new insight into the potential effect of FA on microbial denitrification metabolism process and even nitrogen cycle in nature niches.
研究领域:
环境科学
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微生物学
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生物地球化学
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DOI
PubMed
高影响因子
BIOPROCESS AND BIOSYSTEMS ENGINEERING
·
2016
·Vol. 39
·期 4
·603-611
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IF 1.901
Zhang Jian-Dong,Cui Zhi-Mei,Fan Xiao-Jun,Wu Hua-Lei,Chang Hong-Hong
Two uncharacterized nicotinamide adenine dinucleotide (NADH) oxidases (named as LpNox1, LpNox2) from Lactobacillus pentosus ATCC 8041 were cloned and overexpressed in Escherichia coli BL21 (DE3). The sequence analysis revealed that the two enzymes are water-forming Noxs with 64 % and 52 % identity to LbNox from Lactobacillus brevis DSM 20054. The optimal pH and temperature of the purified LpNox1 and LpNox2 were 7.0 and 8.0 and 35 and 40 °C, respectively, with K
M of 99.0 μM (LpNox1) and 27.6 μM (LpNox2), and yielding catalytic efficiency k
cat / K
M of 1.0 and 0.2 μM −1 s −1 , respectively. Heat inactivation studies revealed that the two enzymes are relatively instable. The application of LpNox1 for the regeneration of NAD + was demonstrated by coupling with a glycerol dehydrogenase-catalyzed oxidation of glycerol to 1,3-dihydroxyacetone. The characteristics of the LpNox1 could prove to be of interest in industrial application such as NAD + regeneration in dehydrogenase-catalyzed oxidations.
研究领域:
生物化学
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微生物学
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2016
·
IF 7.476
Shanying Han,Tianyu Du,Hui Jiang,Xuemei Wang
A self-assembly composite of graphene-pyrroloquinoline quinone (PQQ) was fabricated and modified on glassy carbon electrode (GCE) for sensitive detection of nicotinamide adenine dinucleotide (NADH). Chitosan (CTS) was applied to disperse graphene to form a stable robust film on GCE. A synergistic effect between PQQ and graphene was observed during the electrocatalytic oxidation of NADH, with about 260 mV reduction in the oxidation potential and 2.5-fold increase in the oxidation current compared with those on the bare GCE. The electrochemical sensors based on the modified electrodes allowed the detection of NADH with a good linear dependence from 0.32 to 220 µM with a high sensitivity of 0.421 µA µM −1 cm −2 and a low detection limit of 0.16 µM (S/N=3). It could also eliminate the interference of electroactive substances like ascorbic acid (AA), uric acid , and dopamine and its derivatives. The outstanding performances of graphene-PQQ/CTS composite capable of improving the electrical conductivity and accelerating the electron transport suggested its promising applications for design of different graphene based composites used in electrochemical sensing and energy fields.
研究领域:
生物化学
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材料科学
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分析化学
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DOI
PubMed
高影响因子
ENVIRONMENTAL RESEARCH
·
2020
·
IF 5.715
Zhen-Ni Yang,Ya-Nan Hou,Bo Zhang,Hao-Yi Cheng,Yang-Chun Yong,Wen-Zong Liu,Jing-Long Han,Shuang-Jiang Liu,Ai-Jie Wang
Biologically synthesized palladium nanoparticles (bio-Pd) have attracted considerable interest as promising green catalysts for environmental remediation. However, the mechanisms by which microorganisms produce bio-Pd remain unclear. In the present study, we investigated the roles of Shewanella oneidensis MR-1 and its NADH dehydrogenases and hydrogenases (HydA and HyaB) in bio-Pd production using formate as the electron donor. The roles of NADH dehydrogenases and hydrogenases were studied by inhibiting NADH dehydrogenases and using hydrogenase mutants (Δ hydA , Δ hyaB , and Δ hydA Δ hyaB ), respectively. The results showed ~97% reduction of palladium by S. oneidensis MR-1 after 24 h using 250 μM palladium and 500 μM formate. Electron microscopy images showed the presence of bio-Pd on both the outer and cytoplasmic membranes of S. oneidensis MR-1. However, the inhibition of NADH dehydrogenases in S. oneidensis MR-1 resulted in only ~61% reduction of palladium after 24 h, and bio-Pd were not found on the outer membrane. The mutants lacking one or two hydrogenases removed 91–96% of palladium ions after 24 h and showed more cytoplasmic bio-Pd but less periplasmic bio-Pd. To the best of our knowledge, this is the first study to demonstrate the role of NADH dehydrogenases of S. oneidensis MR-1 in the formation of bio-Pd on the outer membrane. It also demonstrates that the hydrogenases (especially HyaB) of S. oneidensis MR-1 contribute to the formation of bio-Pd in the periplasmic space. This study provides mechanistic insights into the production of biogenic metal nanoparticles towards their possible use in industrial and environmental applications.
研究领域:
微生物学
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环境科学
·
纳米生物学
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产品关联
DOI
PubMed
高影响因子
RARE METALS
·
2025
·
IF 11
Xue Hong-Jin,Zhang Meng,Li Yong-Qing,Liu Chao-Ran,Ma Xin-Yu,He Qin-Bin,Wang Yin-Chuan,Chen Jin-Xing,Qiu Jian-Feng
Constructing high-performance nanozymes for specific biomolecules is crucial but challenging for practical applications and fundamental research. Herein, through the examination of the catalytic reaction paths of natural nicotinamide adenine dinucleotide (NADH) oxidase (NOX), a novel and efficient single-atom rhodium catalyst (Rh 1 /NC) was developed to mimic NOX. The Rh 1 /NC demonstrated the ability to catalyze the dehydrogenation of NADH and transfer electrons to O 2 to generate H 2 O 2 through the typical two-electron pathway. Furthermore, our findings revealed that Rh 1 /NC exhibits the ability to catalyze the conversion of produced H 2 O 2 into OH under mildly acidic conditions. This process amplifies the oxidation of NADH, showcasing NADH peroxidase-like activity (NPx-like). As a paradigm, this unique dual enzyme-like property of Rh 1 /NC with a positive feedback effect holds significance in disrupting cancer cellular homeostasis. Rh 1 /NC can effectively consume NADH via cascade biocatalytic reactions within cancer cells, further triggering the elevation of reactive oxygen species (ROS), leading to impaired oxidative phosphorylation and decreased mitochondrial membrane potential, thus damaging the adenosine triphosphate (ATP) synthesis. The resulting 'domino effect' interferes with the energy metabolism homeostasis of cancer cells, ultimately promoting cell apoptosis. This study provides potential guidance for the rational design of materials with greater capabilities. Graphical abstract
研究领域:
生物化学
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纳米医学
·
肿瘤学
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产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2018
·
IF 5.667
Shuyun Zhu,Cuihua Lei,Jing Sun,Xian-En Zhao,Xiao Wang,Xiaolu Yan,Wei Liu,Hua Wang
In the present study, a novel colorimetric sensing platform was constructed for probing the NAD + /NADH-dependent biocatalytic transformations. Manganese dioxide (MnO 2 ) nanosheets as an oxidase-mimicking nanomaterial could directly oxidize 3,3′,5,5′-tetramethylbenzidine (TMB) into oxTMB without the need of H 2 O 2 . Importantly, it was found that NADH could easily trigger the decomposition of MnO 2 nanosheets, causing the decrease of solution absorbance. This is demonstrated by the enzyme catalysis reactions of two dehydrogenase models, ethanol dehydrogenase (ADH) and lactate dehydrogenase (LDH), where NADH is formed and consumed, respectively, in their physiological enzymatic reactions. A MnO 2 -TMB sensing platform was thereby proposed for the analysis of their catalytic reaction substrates including ethanol and pyruvic acid. The detection limits of ethanol and pyruvic acid can reach 5.0 μM and 100 nM, respectively. The developed optical biosensing platform can be used to develop other dehydrogenase-based biosensors followed by monitoring their substrates.
研究领域:
生物化学
·
纳米生物学
·
分析化学
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产品关联
DOI
高影响因子
Inorganic Chemistry Frontiers
·
2020
·Vol. 7
·期 13
·2434-2442
·
IF 5.958
Changchao Jia,Wenjuan Hu,Yuanyuan Zhang,Chao Teng,Zupeng Chen,Jian Liu
The development of a metal-free photoelectrode film is of great significance. Herein, graphitic carbon nitride (g-C3N4) nanosheets with a concentration of up to 36 mg mL−1 were first obtained on a large scale using a wet ball-milling method. The obtained g-C3N4 nanosheets exhibit 6- and 8-times higher activity in terms of photocatalytic H2 evolution and nicotinamide adenine dinucleotide (NADH) regeneration than bulk g-C3N4, respectively. Furthermore, a uniform g-C3N4 film electrode was fabricated via the interfacial self-assembly of nanosheets at the air/water interface, which can be transferred onto various substrates. By coupling with graphene nanosheets, a g-C3N4/graphene hybrid film electrode was assembled at the interface, showing improved photoelectrochemical coenzyme NADH regeneration efficiency. The photoelectrochemical system uses water as the electron donor, which avoids the drawback of using additional sacrificial agents. This work presents a novel and facile method to prepare high-quality g-C3N4 hybrid films, and also provides a sustainable route for renewable energy conversion and biocatalytic applications.
研究领域:
材料科学
·
生物化学
·
可再生能源
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产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）