JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2023
·
IF 6.1
Mu-Qiang Wang,Zhi-Neng You,Bing-Yi Yang,Zi-Wei Xia,Qi Chen,Jiang Pan,Chun-Xiu Li,Jian-He Xu
Enzymatic synthesis of ursodeoxycholic acid (UDCA) catalyzed by an NADH-dependent 7β-hydroxysteroid dehydrogenase (7β-HSDH) is more economic compared with an NADPH-dependent 7β-HSDH when considering the much higher cost of NADP+/NADPH than that of NAD+/NADH. However, the poor catalytic performance of NADH-dependent 7β-HSDH significantly limits its practical applications. Herein, machine-learning-guided protein engineering was performed on an NADH-dependent Rt7β-HSDHM0 from Ruminococcus torques. We combined random forest, Gaussian Naïve Bayes classifier, and Gaussian process regression with limited experimental data, resulting in the best variant Rt7β-HSDHM3 (R40I/R41K/F94Y/S196A/Y253F) with improvements in specific activity and half-life (40 °C) by 4.1-fold and 8.3-fold, respectively. The preparative biotransformation using a “two stage in one pot” sequential process coupled with Rt7β-HSDHM3 exhibited a space–time yield (STY) of 192 g L–1 d–1, which is so far the highest productivity for the biosynthesis of UDCA from chenodeoxycholic acid (CDCA) with NAD+ as a cofactor. More importantly, the cost of raw materials for the enzymatic production of UDCA employing Rt7β-HSDHM3 decreased by 22% in contrast to that of Rt7β-HSDHM0, indicating the tremendous potential of the variant Rt7β-HSDHM3 for more efficient and economic production of UDCA.
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高影响因子
ACS Applied Materials & Interfaces
·
2014
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IF 5.9
Li Zhang,Hetong Qi,Jie Hao,Lifen Yang,Ping Yu,Lanqun Mao
Developing water-stable and adaptive supramolecular materials is of great importance in various research fields. Here, we demonstrate a new kind of water-stable, adaptive, and electroactive supramolecular ionic materials (SIM) that is formed from the aqueous solutions of imidazolium-based dication and dianionic dye (i.e., 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), ABTS) through ionic self-assembly. The formed SIM not only shows good thermostability and unique optical and electrochemical properties that are raised from precursors of the SIM, but also exhibits good water-stability, salt-stability, and adaptive encapsulation properties toward some heterocyclic cationic dye molecules. UV–vis and FT-IR results demonstrate that this encapsulation property is essentially based on the electrostatic interactions between the guest dye molecules and ABTS in the SIM. The application of the SIM prepared here is illustrated by the development of a new electrochemical sensor for NADH sensing at a low potential. This study not only opens a new avenue to the preparation of the supramolecular materials, but also provides a versatile platform for electrochemical (bio)sensing.
研究领域:
材料科学
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生物传感
·
化学
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RSC Advances
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2016
·Vol. 6
·期 51
·45829-45834
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IF 3.289
Xia Lin,Qinghong Wang,Shu Zhu,Juanjuan Xu,Qiao Xia,Yingzi Fu
In this article, a sensitive and stereo-selective biosensor for L-glutamic acid (L-Glu) based on the electrochemiluminescence (ECL) of Ru(bpy)32+ has been designed by applying L-glutamic dehydrogenase (GLDH) for enzymatic generation of NADH in situ. The novel ECL Glu biosensor was prepared by immobilizing GLDH on the modified electrodes with Ru(bpy)32+–platinum nanoparticles (Ru–PtNPs) and reduced graphene oxide–multiwall carbon nanotubes–Nafion (rGO–MWCNT–Nf). Fourier transform infrared spectroscopy (FT-IR), electrochemical impedance spectroscopy (EIS), scanning electron microscopy (SEM) and cyclic voltammetry (CV) were employed to investigate the characteristics of different materials. In addition, the ECL technology was adopted to explore the interactions between the prepared Glu biosensor and L- or D-Glu. A remarkable difference of ECL intensity was observed when the proposed biosensor interacted with L- and D-Glu, respectively, and good responses were obtained from L-Glu in a range from 1.0 × 10−7 M to 5.0 × 10−3 M with a low detection limit of 3.3 × 10−8 M (S/N = 3). The developed ECL biosensor showed satisfying sensitivity, selectivity, reproducibility and stability, therefore, it might provide a new perspective for the recognition and detection of L-Glu.
研究领域:
生物化学
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分析化学
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高影响因子
ADVANCED MATERIALS
·
2026
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IF 29.1
Jingtao Liu,Xun Guo,Nan Wang,Xingyu Zhang,Xiaoting Wang,Guangru Li,Xumin Hou,Luodan Yu,Yu Chen,Ruogu Li
Myocardial ischemia/reperfusion (I/R) injury is a globally recognized pathological condition that results in life-threatening organ dysfunction. Supplementation with nicotinamide adenine dinucleotide (NAD + ) can improve myocardial energy metabolism, alleviate inflammation, and mitigate I/R injury. However, its clinical applicability is constrained by the poor efficacy and delivery limitations of current NAD + supplementation strategies. Here, we design a dual single-atom nanozyme with nicotinamide adenine dinucleotide (NADH) oxidase-mimicking activity for in situ NAD + biotransformation and I/R injury treatment. Upon M2 macrophage membrane coating, the nanozyme selectively targets ischemic myocardium, facilitating in situ NAD + biotransformation by eliminating excess NADH, while concurrently scavenging reactive oxygen species, restoring mitochondrial bioenergetics, attenuating inflammation, and inhibiting apoptosis. Mechanistic investigations demonstrate that the nanozyme activates the NAD + -SIRT1 axis, enhancing ATP biosynthesis and oxidative phosphorylation, while suppressing TNF signaling and NF-κB pathways. Collectively, the engineered NOX-like nanozyme offers a novel therapeutic strategy for myocardial I/R injury by modulating both mitochondrial bioenergetics and oxidative stress defense pathways through restoring NAD + /NADH homeostasis.
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CHEMICAL ENGINEERING JOURNAL
·
2022
·
IF 16.744
Lihang Yan,Guanhua Liu,Jianqiao Liu,Jing Bai,Yaoxuan Li,Huaxun Chen,Liya Zhou,Jing Gao,Yanjun Jiang
The utilization of CO 2 has become an important issue aiming to the achievement of carbon neutrality. The enzyme electrocatalytic CO 2 reduction to produce carbonaceous fuels has been increasingly appealing. While the low CO 2 aqueous solubility severely limits the CO 2 reduction efficiency, and the expensive cofactor restricts the application potential. In this study, microporous UiO-66-NH 2 was firstly converted into hierarchically porous structure (HP-UiO-66-NH 2 ) containing both micropores and mesopores to simultaneously enhance the enrichment of CO 2 and the immobilization of formate dehydrogenase by optimizing the pore structure. The relative contents of the protein secondary structures were similar after immobilization. The mechanism of enzyme electrocatalytic CO 2 reduction was proposed. The enzyme stability was highly enhanced after immobilization and the CO 2 reduction was enhanced by the enzyme electrocatalytic system through the combination of CO 2 enrichment and electrocatalytic NADH regeneration. The optimized catalytic system can obtain formate yield of 1.826 mM in 3 h (5.57 times higher than the free enzyme system) with a generation rate of 6086.7 μmol g cat ‑1 h ‑1 .
研究领域:
生物化学
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环境科学
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材料科学
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Small
·
2023
·
IF 13.3
Rui Pei,Jing Liu,Chuanyong Jing,Min Zhang
The reduction of carbon dioxide to valuable chemicals through enzymatic processes is regarded as a promising approach for the reduction of carbon dioxide emissions. In this study, an in vitro multi-enzyme cascade pathway is constructed for the conversion of CO 2 into dihydroxyacetone (DHA). This pathway, known as FFFP, comprises formate dehydrogenase (FDH), formaldehyde dehydrogenase (FaldDH), formolase (FLS), and phosphite dehydrogenase (PTDH), with PTDH serving as the critical catalyst for regenerating the coenzyme NADH. Subsequently, the immobilization of the FFFP pathway within the hydrogen-bonded organic framework (HOF-101) is accomplished in situ. A 1.8-fold increase in DHA yield is observed in FFFP@HOF-101 compared to the free FFFP pathway. This enhancement can be explained by the fact that within FFFP@HOF-101, enzymes are positioned sufficiently close to one another, leading to the elevation of the local concentration of intermediates and an improvement in mass transfer efficiency. Moreover, FFFP@HOF-101 displays a high degree of stability. In addition to the establishment of an effective DHA production method, innovative concepts for the tailored synthesis of fine compounds from CO 2 through the utilization of various multi-enzyme cascade developments are generated by this work.
研究领域:
生物化学
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环境科学
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材料科学
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SENSORS AND ACTUATORS B-CHEMICAL
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2025
·
IF 7.7
Li-Jun Zhao,Qing Zhou,Na Zhou,Shixin Zhang,Yue Yang,Chao Shi,Yanhong Li,Feng Luan,Chunyuan Tian,Peng Xiao,Xuming Zhuang
Here, we developed the cyclodextrin-organoiridium-functionalized magnetic nanocatalyst MBs@CD@Ir by assembling highly active adamantane-functionalized pyridinamide organoiridium complexes PicPh-Ada-Ir and cyclodextrin-modified magnetic nanoparticles MBs@CD , which could efficiently catalyze quinone hydrogenation using the cofactor NADH as a hydrogen source under physiological conditions. Furthermore, by combining the developed catalysts MBs@CD@Ir and the quinone hydrogenation-triggered Rhodamine 110-type fluorescence probe Q-Rho 110 , we successfully established a sensitive cofactor NADH fluorescence sensor, which showed superior hydrogenation performance for quinone when NADH was used as a hydrogen source. The established sensor presented excellent detection performance for NADH with a limit of detection (LOD) of 4.18 nM and was successfully applied to NADH detection in Escherichia coli fermentation broth. The established sensor has successfully expanded the application of organometallic catalysts in biosensing and provides a new approach for the design of other sensors.
研究领域:
生物化学
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分析化学
·
催化科学
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ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Ying Li,Xiwen Zhang,Yiyu Wang,Xue Mu,Yuanbo Tu,Peng Sun,Yaolong Wang,Gongjun Yang,Lifeng Kang,Chunyong Wu,Junying Zhang
The insertion of a single nanoelectrode into the cell or fluorescence detection has enabled the nondestructive tracking of intracellular biomarkers. However, the reliance on extremely expensive micromanipulators or intricate fluorescent probes has limited their widespread application. To address this challenge, we propose a novel strategy that employs modified multiwalled carbon nanotubes (MWCNTs) as artificial transmembrane electron tunnels. These MWCNTs can be spontaneously and nondestructively semiembedded within cells while exhibiting remarkable conductivity. When the photosensitive electrodes on the cell surface are illuminated, photogenerated electron–hole pairs are produced. The holes can enter the cell through the nanowires, thus promoting oxidation reactions with target molecules and generating real-time electrical signals for monitoring intracellular biomarkers. Meanwhile, the engineered recognition element on the nanowire transitions to an excited state and subsequently returns to the ground state through electrochemiluminescence (ECL), enabling in situ visualization of intracellular biomarkers. This groundbreaking approach not only eliminates the need for micromanipulators and fluorescent probes but also enables simultaneous electrical and optical monitoring of intracellular biomarker levels, significantly reducing false-positive risks. This innovation was validated to be feasible using NADH as an intracellular target molecule, marking a shift in intracellular sensing strategies.
研究领域:
生物医学工程
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分析化学
·
纳米技术
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ELECTRONIC JOURNAL OF BIOTECHNOLOGY
·
2020
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IF 2.894
Longxing Wang,Wenjun Zhu,Zhen Gao,Hua Zhou,Fei Cao,Min Jiang,Yan Li,Honghua Jia,Ping Wei
Background L- tert -Leucine has been widely used in pharmaceutical, chemical, and other industries as a vital chiral intermediate. Compared with chemical methods, enzymatic methods to produce L- tert -leucine have unparalleled advantages. Previously, we found a novel leucine dehydrogenase from the halophilic thermophile Laceyella sacchari ( Ls LeuDH) that showed good thermostability and great potential for the synthesis of L- tert -leucine in the preliminary study. Hence, we manage to use the Ls LeuDH coupling with a formate dehydrogenase from Candida boidinii ( Cb FDH) in the biosynthesis of L- tert -leucine through reductive amination in the present study. Result The double-plasmid recombinant strain exhibited higher conversion than the single-plasmid recombinant strain when resting cells cultivated in shake flask for 22 h were used. Under the optimized conditions, the double-plasmid recombinant E. coli BL21 (pETDute-FDH-LDH, pACYCDute-FDH) transformed 1 mol·L -1 trimethylpyruvate (TMP) completely into L- tert -leucine with greater than 99.9% ee within 8 h. Conclusions The Ls LeuDH showed great ability to biosynthesize L- tert -leucine. In addition, it provided a new option for the biosynthesis of L- tert -leucine. How to cite Wang L, Zhu W, Gao Z, et al. Biosynthetic L- tert -leucine using Escherichia coli co-expressing a novel NADH-dependent leucine dehydrogenase and a formate dehydrogenase. Electron J Biotechnol 2020;47. https://doi.org/10.1016/j.ejbt.2020.07.001
研究领域:
生物技术
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微生物学
·
酶学
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ANALYTICAL CHEMISTRY
·
2023
·
IF 7.4
Dan Sun,Guohua Qi,Xuan Yi,Hongyan Zhu,Yongdong Jin
Studying the oxidative stress, especially the reactive oxygen species (ROS) response of ferroptosis, is crucial for the diagnosis and treatment of cancer based on ferroptosis. However, reliable quantitative analysis of intracellular ROS in cancer treatment for drug screening is still a challenge. Herein, a superior ratiometric SERS nanoprobe was developed for in situ, real-time, and highly sensitive detection of content variation of H2O2 within living cells. The SERS nanoprobe was prepared by coassembly of the internal standard molecule p-mercaptobenzonitrile and the reporter molecule p-mercaptophenylboronic acid on the surface of gold nanoparticles, used for synergistic calibration and detection of H2O2, which enables reliable detection of the true content of intracellular H2O2 without the interference of other substances in cells. Based on the nanoprobe, we found that the level of intracellular H2O2 of cancer cells was increased after the nicotinamide adenine dinucleotide (NADH) treatment, with a dose-dependence to the concentration of NADH. High doses of NADH (above 20 mM) can induce cell death by means of ferroptosis associated with the level elevation of intracellular lipid hydroperoxides. This study highlights the potential of the SERS nanoprobe for tracking content variation of cellular H2O2 and understanding its roles in screening new anticancer drugs.
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Nano Energy
·
2020
·
IF 16.602
Wengang Liu,Wenjuan Hu,Lijun Yang,Jian Liu
By analogy to mono-nuclear metalloenzymes , single-atom dispersed catalysts (SACs) demonstrate very promising performance for many conventional heterogeneous and homogeneous reactions. However, the prospect of integrating SACs with enzymatic catalysis is still unexplored. Herein, atomically dispersed Co sites on ultrathin two-dimensional (2D) carbon nitride nanosheets (Co 1 /C 3 N 4 ) were constructed and applied for efficiently photochemical cofactor (NADH) regeneration. The NADH regeneration yield reached as high as 99%. Subsequently, the in-situ regenerated-NADH triggered alcohol dehydrogenase for enzymatic reduction of aldehyde where 100% alcohol yield was achieved. Using atomic spectroscopy characterization and theoretical calculation, the genuine active sites of Co 1 /C 3 N 4 catalyst in reaction system were revealed in the form of CoN 3 - Cp* moieties ( Cp* , 1,2,3,4,5-pentamethylcyclopentadiene). Such a well-defined atomic catalyst opens a new avenue for the organic transformation application of single-atom catalysts in enzyme catalysis.
研究领域:
催化
·
材料科学
·
酶工程
·
光化学
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高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2020
·
IF 4.192
Tian Li,Guo-Song Liu,Wei Zhou,Min Jiang,Yu-Hong Ren,Xin-Yi Tao,Min Liu,Ming Zhao,Feng-Qing Wang,Bei Gao,Dong-Zhi Wei
Squalene has wide applications in the food and pharmaceutical industries. Engineering microbes to produce squalene is a promising alternative for traditional production approaches. In this study, squalene production was enhanced to 978.24 mg/L through stepwise overexpression of the enzymes that catalyze acetyl-CoA to squalene. Subsequently, to increase the activity of HMG-CoA reductase and alleviate the high dependence on NADPH, the HMG-CoA reductase (NADH-HMGR) from Silicibacter pomeroyi, highly specific for NADH, was introduced, which increased squalene production to 1086.31 mg/L. Native ethanol dehydrogenase ADH2 and acetaldehyde dehydrogenase ADA from Dickeya zeae were further overexpressed, which enhanced the capability to utilize ethanol for squalene synthesis and endowed the engineered strain with greater adaptability to high ethanol concentrations. Finally, a remarkable squalene production of 9472 mg/L was obtained from ethanol via carbon source-controlled fed-batch fermentation. This study will greatly accelerate the process of developing microbial cell factories for squalene production.
研究领域:
生物技术
·
代谢工程
·
微生物生物技术
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高影响因子
危险品化学品经营许可证（带存储）