ACS Catalysis
·
2024
·
IF 11.3
Zaixiang Xu,Fengcang Zhou,Hanchi Chen,Jianguo Wang
The regeneration of NAD(P)H and its analogues is crucial for biocatalytic processes. However, despite the efficiency of enzymatic catalysis in regenerating NAD(P)H, the sustainability of enzymes is often compromised, particularly under extreme catalytic conditions. Moreover, artificial cofactors may present advantages in certain reactions due to their stability and versatility, yet the substrate specificity of enzymes poses significant challenges to their regeneration through the enzymatic method. Therefore, it is imperative to develop a highly stable regeneration method that can be adapted to both natural and artificial cofactors. In this work, employing potassium-ion-doped carbon nitride (ionCN-0.2) as a catalyst not only achieves high-efficiency photocatalytic regeneration of NAD(P)H, comparable to that of glucose dehydrogenase (GDH), but also a remarkable ability to regenerate nicotinamide analogues. This enhanced performance stems from the tunable negative ζ-potential, which effectively adsorbs the positively charged [Cp*Rh(bpy)H2O]2+ mediator, resulting in enhanced regeneration kinetics of the nicotinamide moiety. The catalyst demonstrates superior performance compared to the reported systems; the optimal regeneration rate reaches 0.55 mmol L–1 gcat–1 min–1 and approaches enzymatic regeneration efficiency. Expanding the reaction conditions to a wider temperature and pH range also confirms the effectiveness and sufficient stability of this photocatalytic system, offering a promising strategy for stable cofactor regeneration in biocatalytic processes.
研究领域:
生物化学
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材料科学
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催化化学
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DOI
高影响因子
BIOMATERIALS
·
2021
·
IF 12.479
Yang Tian,Wen-Li Jiang,Wen-Xin Wang,Guo-Jiang Mao,Yongfei Li,Chun-Yan Li
The reduced coenzymes (NADH and NADPH) are an important product in energy metabolism and closely related to the occurrence and development of cancer. So it is necessary to use a powerful detection tool to visualize NAD(P)H in energy metabolism of tumor cells and find a new strategy to improve cancer treatment based on NAD(P)H. Herein, a novel multifunctional probe (Cy-N) is synthesized with good near-infrared fluorescence (NIRF) response to NAD(P)H and the photoacoustic (PA) and photothermal properties are successfully activated by NAD(P)H. The probe is successfully applied in visualizing NAD(P)H in energy metabolism of tumor cells and imaging NAD(P)H in bacteria. Moreover, the probe can be used to image NAD(P)H in energy metabolism of tumor-bearing mice by dual-modal imaging (NIRF and PA). More importantly, in terms of the role of NAD(P)H in energy metabolism, the photothermal therapy (PTT) is activated by NAD(P)H and a novel strategy of enhanced PTT is proposed by injecting glucose. As far as we know, this is the first probe to detect NAD(P)H in energy metabolism through dual-modal imaging, and also the first probe to activate PTT based on NAD(P)H, which will provide important information of the diagnosis and treatment of cancer.
研究领域:
生物化学
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医学影像学
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癌症治疗
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL BIOCHEMISTRY
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2016
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IF 2.243
Wei Zhang,Min Zhu,Feng Wang,Danhui Cao,Jennifer Jin Ruan,Weike Su,Benfang Helen Ruan
Glutamate dehydrogenase (GDH) catalyzes the oxidative deamination of L-glutamate and is important for several biological processes . For GDH inhibitor screening, we developed a novel mono-sulfonated tetrazolium salt (EZMTT), which can be synthesized using H 2 O 2 oxidation and purified easily on silica gel in large quantities. The EZMTT detection method showed linear dose responses to NAD(P)H, dehydrogenase concentration and cell numbers. In E. coli GDH assay, the EZMTT method showed excellent assay reproducibility with a Z factor of 0.9 and caused no false positives in the presence of antioxidants (such as BME). Using the EZMTT-formazan-NAD(P)H system, we showed that EGCG is a potent E. coli GDH inhibitor (IC 50 45 nM) and identified that Ebselen , a multifunctional thioredoxin reductase inhibitor, inactivated E. coli GDH (IC 50 213 nM). In cell-based assays at 0.5 mM tetrazolium concentration, EZMTT showed essentially no toxicity after a 3-day incubation, whereas 40% of inhibition was observed for WST-8. In conclusion, EZMTT is a novel tetrazolium salt which provides improved features that are suitable for dehydrogenases and real-time cell-based high-throughput screening (HTS).
研究领域:
生物化学
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药物筛选
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细胞生物学
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DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2023
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IF 9.5
Ying Zhang,Bin Wei,Hao Liang
An organometallic complex-catalyzed artificial coenzyme regeneration system has attracted widespread attention. However, the combined use of organometallic complex catalysts and natural enzymes easily results in mutual inactivation. Herein, we establish a rhodium-based metal–organic framework (MOF)-on-MOF difunctional core–shell nanoreactor as an artificial enzymatic NAD(P)H regeneration system. UiO67 as the core is used to capture rhodium molecules for catalyzing NAD(P)H regeneration. UiO66 as the shell is used to specifically immobilize His-tagged lactate dehydrogenase (LDH) and serve as a protection shield for LDH and [Cp*Rh(bpy)Cl]+ to prevent mutual inactivation. A variety of results indicate that UiO67@Rh@UiO66 has good activity in realizing NAD(P)H regeneration. Noteworthily, UiO67@Rh@UiO66@LDH maintains a high activity level even after 10 cycles. This work reports a novel NAD(P)H regeneration platform to open up a new avenue for constructing chemoenzyme coupling systems.
研究领域:
生物化学
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材料科学
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酶工程
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DOI
PubMed
高影响因子
NEW JOURNAL OF CHEMISTRY
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2022
·Vol. 46
·期 13
·6274-6282
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IF 3.925
Xiu Xing,Yan Liu,Ming-Liang Shi,Kun Li,Xin-Yue Fan,Zhong-Liu Wu,Na Wang,Xiao-Qi Yu
Controllable and mild photoenzymatic production of chiral alcohols was realized by coupling a versatile photochemical NAD(P)H regeneration system with (R)- or (S)-selective ketoreductases. The efficiency of NAD(P)H regeneration was improved using a Rhodium functionalized metal organic framework, namely Rh-UiO-67, to adjust and control electron transport and electron utilization. Furthermore, six different ketoreductases could be successfully immobilized on Rh-UiO-67 and combined with the light-driven NAD(P)H regeneration system to produce chiral aryl alcohols. Various chiral alcohols with complementary (R)- and (S)-conformations can be constructed by this method with high yields (97%) and excellent stereoselectivity (>99% ee).
研究领域:
化学
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生物化学
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催化
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有机合成
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产品关联
DOI
高影响因子
Chemical Science
·
2019
·Vol. 10
·期 35
·8179-8186
·
IF 9.556
Xiaohong Pan,Yuehui Zhao,Tingting Cheng,Aishan Zheng,Anbin Ge,Lixin Zang,Kehua Xu,Bo Tang
Reductive stress, the opposite of oxidative stress, represents a disorder in the redox balance state which is harmful to biological systems. For decades, the role of oxidative stress in tumor therapy has been the focus of attention, while the effects of reductive stress have been rarely studied. Here, we report the anti-cancer effects of reductive stress induced by three natural antioxidants (resveratrol, curcumin and celastrol). Considering the fact that the solid tumor microenvironment suffers from hypoxia, we performed cell experiments under hypoxic conditions. In order to observe the reductive stress, we first developed an ultrasensitive fluorescent probe (TCF-MQ) for specifically imaging NAD(P)H which is a marker of reductive stress. TCF-MQ responded to NAD(P)H rapidly and exhibited high sensitivity with a detection limit of 6 nM. With the help of TCF-MQ, we found that upon the treatment of HepG2 cells with pharmacological doses of three natural antioxidants under hypoxic conditions, high levels of NAD(P)H were produced before cell death. The excess NAD(P)H resulted in reductive stress instead of oxidative stress. In contrast, under normoxic conditions, there was no reductive stress involved in the process of cell death induced by three natural antioxidants. Therefore, we hypothesize that the mechanism of cancer cell death induced by natural antioxidants under hypoxia should be attributed to the reductive stress.
研究领域:
生物化学
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肿瘤学
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药理学
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DOI
PubMed
高影响因子
Nature Communications
·
2023
·Vol. 14
·期 1
·1-12
·
IF 16.6
Liu Jianming,Zhang Han,Xu Yingying,Meng Hao,Zeng An-Ping
The use of gaseous and air-captured CO 2 for technical biosynthesis is highly desired, but elusive so far due to several obstacles including high energy (ATP, NADPH) demand, low thermodynamic driving force and limited biosynthesis rate. Here, we present an ATP and NAD(P)H-free chemoenzymatic system for amino acid and pyruvate biosynthesis by coupling methanol with CO 2 . It relies on a re-engineered glycine cleavage system with the NAD(P)H-dependent L protein replaced by biocompatible chemical reduction of protein H with dithiothreitol. The latter provides a higher thermodynamic driving force, determines the reaction direction, and avoids protein polymerization of the rate-limiting enzyme carboxylase. Engineering of H protein to effectively release the lipoamide arm from a protected state further enhanced the system performance, achieving the synthesis of glycine, serine and pyruvate at g/L level from methanol and air-captured CO 2 . This work opens up the door for biosynthesis of amino acids and derived products from air.
研究领域:
生物技术
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化学工程
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生物化学
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2025
·
IF 7.7
Bin Jiang,Jiayun Guan,Yilin Lv,Chaojie Yang,Minfeng Yang,Xinyu Shen,Bin Xu,Qi Wang,Yuling Qin,Li Wu
The cholinergic hypothesis constitutes a pivotal theoretical framework for elucidating the pathological mechanisms underpinning Alzheimer's disease (AD). Quantitative assessment of cholinesterases (ChEs) and NAD(P)H levels in neuronal cells, particularly within the septo-hippocampal pathway, is imperative for the comprehensive study of the cholinergic system. In this study, we designed and synthesized a dual-channel, dual-enzyme-responsive fluorescent indicator that was chemically optimized for detecting levels of ChEs and NAD(P)H in neuronal cells. Biological experiments demonstrated that this indicator can sensitively and reliably visualize the dynamics of intracellular ChEs and NAD(P)H following stimulation. The corresponding ChEs activities and NAD(P)H concentrations demonstrated a linear correlation with fluorescence intensity, reaching equilibrium within a few minutes. This innovative and precise technique for visualizing ChEs and NAD(P)H dynamics with high spatial resolution during the progression of AD is expected to serve as a robust tool for investigating alterations in the cholinergic nervous system, including changes in acetylcholine neurotransmission and neuronal energy metabolism.
研究领域:
神经科学
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生物化学
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分子生物学
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医学研究
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荧光成像
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产品关联
DOI
高影响因子
ACS Sustainable Chemistry & Engineering
·
2025
·
IF 7.3
Weiwei Liu,Zichen Yu,Zhichao Chen,Ling Ma,Yanghao Liu,Zhen Zhang,Qingyang Xu
5-Hydroxytryptamine (5-HT), also known as serotonin, plays a critical role in mood regulation, insomnia treatment, and free radical scavenging. Currently, 5-HT production primarily relies on plant extraction, which is challenged by limited raw materials and environmental concerns, emphasizing the need for innovative green production strategies. In this study, we explored a high-performance producer for biosynthesizing 5-HT in Escherichia coli. First, we constructed and optimized the 5-HT biosynthetic pathway to convert L-tryptophan (l-Trp) to 5-HT, avoiding the accumulation of tryptamine and 5-hydroxytryptophan. Moreover, precursor enrichment eliminated the need for l-Trp supplementation and dynamic downregulation of pyruvate kinase balanced growth and production. Further, the metabolic flux between the tetrahydrobiopterin (BH4) module and the hydroxylation-decarboxylation module was optimized, reducing l-Trp accumulation. A novel “GXAA” system was implemented to enhance intracellular NAD(P)H supply, including (1) expression of gdhbsu (glucose dehydrogenase) from Bacillus subtilis, gapXfbr (mutated glyceraldehyde-3-phosphate dehydrogenase) from Corynebacterium glutamicum, and pntAB (pyridine nucleotide transhydrogenase); (2) knockout of ldhA (lactate dehydrogenase). Finally, transcriptomic analysis under high-5-HT stress revealed a potential 5-HT transporter, YhjX. The resulting strain WW15 produced 15.5 g/L 5-HT and a yield of 0.063 g/g glucose through a two-stage fed-batch fermentation, representing the highest titer and yield reported to date.
研究领域:
生物化学
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合成生物学
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微生物学
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DOI
高影响因子
CHEMISTRY-A EUROPEAN JOURNAL
·
2023
·
IF 4.3
Yuan-Yang Guo,Ze-Hua Tian,Yu-Chen Han,Dandan Ma,Tianju Shao,Zhiyong Jiang
Graphical Chemically stable Hantzsch ester is discovered to be an efficient and economical NAD(P)H mimic as sacrificial electron source in in vitro bioconversions. Biocatalysis has emerged as a valuable and reliable tool for industrial and academic societies, particularly in fields related to bioredox reactions. The cost of cofactors, especially those needed to be replenished at stoichiometric amounts or more, is the chief economic concern for bioredox reactions. In this study, a readily accessible, inexpensive, and bench-stable Hantzsch ester is verified as the viable and efficient NAD(P)H mimic by four enzymatic redox transformations, including two non-heme diiron N -oxygenases and two flavin-dependent reductases. This finding provides the potential to significantly reduce the costs of NAD(P)H-relying bioredox reactions.
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DOI
PubMed
高影响因子
Nanomaterials
·
2024
·Vol. 14
·期 6
·512
·
IF 4.4
Yuan Chen,Jiaxing Wang,Daidi Chen,Boxi Wang,Jinchuan Wu,Rongrong Liu,Qingxin Li
Many studies have been conducted on the microbial reduction of Pd (II) to palladium nanoparticles (Pd-NPs) due to the environmental friendliness, low cost, and the decreased toxicity of Pd (II) ions. In this study, we investigate the reduction mechanism of Pd (II) byBacillus megateriumY-4 through proteomics. The data are available via ProteomeXchange with identifier PXD049711. Our results revealed thatB. megateriumY-4 may use the endogenous electron donor (NAD(P)H) generated by nirB, tdh, and fabG and reductase to reduce Pd (II) to Pd-NPs. The expression levels of fabG, tdh, gudB, and rocG that generate NAD(P)H were further increased, and the number of reduced Pd-NPs was further increased with the exogenous electron donor sodium formate. Endogenous electron mediators such as quinones and flavins inB. megateriumY-4 can further enhance Pd (II) reduction. The findings provided invaluable information regarding the reduction mechanism of Pd (II) byB. megateriumY-4 at the proteome level.
研究领域:
微生物学
·
环境科学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF FOOD SCIENCES AND NUTRITION
·
2019
·
IF 2.792
Yuanfeng Wu,Yuke Shen,Ye Zhu,Jothame Mupunga,Ligen Zou,Chao Liu,Shiwang Liu,Jianwei Mao
After consuming broccoli, isothiocyanates can be produced by the hydrolytic action of myrosinase from plant and/or microbiota. Using male C57BL/6 mice, the present study investigated the effects of broccoli ingestion on the myrosinase-like activity, NAD(P)H:quinone oxidoreductase 1 (NQO1) activity, diversity and composition of the gut microbiota. Compared with the control group, continuously feeding raw or hydrolysed broccoli increased the myrosinase-like activities of the colon and caecum contents, and also improved the NQO1 activity of the colon mucosa. Significant difference between the broccoli and control feeding groups were found. 16S rRNA gene sequencing indicated that broccoli ingestion profoundly affected the composition of the gut bacteria community. The correlation between the gut bacterial community composition and microbiota myrosinase-like activity was also studied. However, one type of glucosinolate, sinigrin, had no effect on these activities, indicating that broccoli component(s), other than glucosinolate, had increased the NQO1 and myrosinase-like activities.
研究领域:
营养学
·
微生物学
·
分子生物学
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DOI
PubMed
高影响因子
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