INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2025
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IF 8.5
Haiyue Chen,Jun Xu,Han Shi,Xinle Liang,Wenjie Wang
The global rise in hyperuricemia (HUA) has driven the quest for safe, non-pharmacological interventions. Food-grade probiotics with HUA-mitigating potential are particularly promising. We isolated a novel strain, SSY-7-2, from traditional fermented wheat pastes, which exhibited robust uric acid (UA) degradation activity. SSY-7-2 showed high average nucleotide identity (ANI) values of 98.76–99.60 % with Priestia megaterium , surpassing the species delineation threshold. Whole-genome sequencing revealed complete UA degradation and purine metabolism pathways. We identified gene SSY_004365 as encoding an FAD-dependent urate hydroxylase (HpmO, E.C. 1.14.13.113), with a Km of 27.36 μM and a Kcat/Km of 5.74 × 10 4 s −1 M −1 . Using UA as the sole nitrogen source, we partially elucidated a conserved HpmO-UraD-HiuH cascade for UA hydrolysis in P. megaterium . Both in vitro and Caenorhabditis elegans models confirmed SSY-7-2's UA degradation activity, safety, and probiotic attributes. These findings expand the bioactivity spectrum of Priestia megaterium and position SSY-7-2 as a promising complementary probiotic for hyperuricemia management.
研究领域:
微生物学
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分子生物学
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代谢疾病
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营养科学
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转化医学
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药理学
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PubMed
高影响因子
NEUROPATHOLOGY AND APPLIED NEUROBIOLOGY
·
2022
·Vol. 48
·期 7
·e12839
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IF 6.25
Xiaolian Ye,Lu Chen,He Wang,Shixiao Peng,Mengjia Liu,Liyang Yao,Yizhi Zhang,Yun Stone Shi,Ying Cao,Jian-Jun Yang,Guiquan Chen
Aims Abundant recent evidence has shown that 3-phosphoinositide-dependent protein kinase 1 (PDK1) is activated in Alzheimer's disease (AD). However, it remains unknown whether inhibition of PDK1 in neurons may affect AD-like pathology in animal models of AD. Here, we aim to examine the effects of specific inactivation of neuronal PDK1 on pathology and behaviour in 5×FAD mice and to identify the underlying molecular mechanisms. Methods The Cre-loxP system was employed to generate Pdk1 cKO/5×FAD mice, in which PDK1 is inactivated in excitatory neurons in the adult forebrain. Cellular and behavioural techniques were used to examine plaque burden, inflammatory responses and spatial working memory in mice. Biochemical and molecular analyses were conducted to investigate relevant mechanisms. Results First, Aβ deposition was massively decreased and gliosis was highly attenuated in Pdk1 cKO/5×FAD mice compared with 5×FAD mice. Second, memory deficits were significantly improved in Pdk1 cKO/5×FAD mice. Third, APP levels were notably decreased in Pdk1 cKO/5×FAD mice. Fourth, mammalian target of rapamycin (mTOR) signalling and ribosome biogenesis were reduced in Pdk1 cKO/5×FAD mice. Conclusions Neuron-specific deletion of PDK1 robustly ameliorates AD-like pathology and improves spatial working memory in 5×FAD mice. We propose that genetic approach to inhibit PDK1 may be an effective strategy to slow AD.
研究领域:
神经科学
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分子生物学
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药理学
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遗传学
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DOI
PubMed
高影响因子
Frontiers in Microbiology
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2018
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IF 4.019
Chen Xin,Tang Hongzhi,Liu Yongdi,Xu Ping,Xue Yong,Lin Kuangfei,Cui Changzheng
3-Hydroxybenzoate 6-hydroxylase, an NADH-dependent flavoprotein, can convert 3-hydroxybenzoate which is an important intermediate in the biodegradation of many aromatic hydrocarbons. 3-Hydroxybenzoate is metabolized by entering the TCA cycle through the gentisate pathway. We found a putative 3HB6H gene from a cluster that potentially encodes for gentisic acid degradation from a halophilic Martelella sp. strain AD-3. The corresponding protein was expressed with an N-terminal His-tag and purified by Ni2+-nitrilotriacetic acid affinity chromatography. The protein showed an overexpressed band of about 46 kDa by SDS–PAGE, and it was also proven that the enzyme contains FAD by absorption spectroscopy and HPLC analysis. The optimal activity of 3HB6H from strain AD-3 was observed in phosphate buffer (pH 8.0) at 37∘C without salinity (NaCl) and metal salts. The Km values of 3-hydroxybenzoate 6-hydroxylase were determined to be 72.6 ± 10.1 μM and 104.1 ± 18.2 μM for 400 μM NADH and 3-hydroxybenzoate, respectively. Site-directed mutagenesis showed that residues 305, 306 and 308 are important for FAD binding. In addition, we found that Tyr221 and Gln305 of 3HB6H from strain AD-3 are involved in substrate binding.
研究领域:
微生物学
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酶学
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DOI
PubMed
高影响因子
Synthetic and Systems Biotechnology
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2024
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IF 4.4
Bing Fu,Meng Chen,Xianfeng Bao,Jiajie Lu,Zhiwen Zhu,Fuyao Guan,Chuyang Yan,Peize Wang,Linglin Fu,Ping Yu
Vitamin B 2 is an essential water-soluble vitamin. For most prokaryotes, a bifunctional enzyme called FAD synthase catalyzes the successive conversion of riboflavin to FMN and FAD. In this study, the plasmid pNEW-AZ containing six key genes for the riboflavin synthesis was transformed into strain R2 with the deleted FMN riboswitch, yielding strain R5. The R5 strain could produce 540.23 ± 5.40 mg/L riboflavin, which was 10.61 % higher than the R4 strain containing plasmids pET-AE and pAC-Z harboring six key genes. To further enhance the production of riboflavin, homology matching and molecular docking were performed to identify key amino acid residues of FAD synthase. Nine point mutation sites were identified. By comparing riboflavin kinase activity, mutations of T203D and N210D, which respectively decreased by 29.90 % and 89.32 % compared to wild-type FAD synthase, were selected for CRISPR/Cas9 gene editing of the genome, generating engineered strains R203 and R210. pNEW-AZ was transformed into R203, generating R6. R6 produced 657.38 ± 47.48 mg/L riboflavin, a 21.69 % increase compared to R5. This study contributes to the high production of riboflavin in recombinant E. coli BL21.
研究领域:
生物工程
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分子生物学
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DOI
PubMed
高影响因子
IEEE JOURNAL OF SELECTED TOPICS IN QUANTUM ELECTRONICS
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2019
·Vol. 25
·期 1
·1-6
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IF 4.681
Xinyi Wang,Yonghui Xie,Maojia Huang,Longfang Yao,Yulan Wang,Yiyan Fei,Jiong Ma,Lan Mi
Fluorescence lifetime measurements are often performed on live as well as fixed cells and tissues. Fixation and mounting processes are routinely used in cellular research or clinical diagnosis. In this paper, the effects of fixation and mounting on the fluorescence lifetime of cellular autofluorescence were studied by fluorescence lifetime imaging microscopy over time. Two endogenous fluorescent fluorophores, reduced nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD), showed different results between live cells and fixed cells. The average lifetime of NADH in live HeLa cells was about 1.02 ns, while maintained about 1.57 ns during the fixation periods of 14 days. The average lifetimes of FAD in live and fixed HeLa cells within 11 days were similar around 1.75 ns but increased to 2.10 ns after 12 days. The free and bound states of the two kinds of fluorophores were further analyzed. It was found that the bound-FAD had two different groups, which was related to the cell division cycle. The effect of mounting medium on fluorescence lifetimes was also studied, indicating glycerol has a negative impact on the fluorescence lifetime compared with neutral balsam.
研究领域:
生物医学工程
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细胞生物学
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DOI
高影响因子
Cell and Bioscience
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2025
·Vol. 15
·期 1
·1-23
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IF 6.2
Ji Hao-Nan,Zhou Hai-Qian,Qie Jing-Bo,Lu Wen-Mei,Gao Hai-Tao,Wu Dan-Hong
Background The identification and intervention of Alzheimer’s Disease (AD) in its early-stage allows for the timely implementation of lifestyle modifications and therapeutic strategies. Although dysregulation of protein expression has been reported in the brain from AD patients and AD animal models, the underlying mechanisms remain poorly understood. N4-acetylcytidine (ac4C), the only known form of RNA acetylation in eukaryotes, has recently been shown to regulate mRNA stability and translation efficiency. However, the dysregulation of ac4C associated with abnormal protein expression levels in the brain of early-stage mouse models of AD remains to be elucidated. Methods This study investigated ac4C modifications, mRNA and protein expression in the hippocampus of 3 and 6-month-old 5×FAD mice, a mouse model of AD, and wild-type (WT) littermates. The multi-omics analysis was performed: acetylated RNA immunoprecipitation followed by next-generation sequencing (acRIP-seq) to identify ac4C mRNAs, deep RNA sequencing (RNA-seq) to quantify mRNA abundance, and label-free quantitative proteomics to assess protein expression levels. In addition, we used acRIP-qPCR, regular qPCR and western blots to verify the ac4C, mRNA and protein levels of some key genes that were identified by the high-throughput assays. Results Proteomic analysis revealed significant change of protein expression in the hippocampus of 3-months-old 5×FAD mice, compared with WT littermates. In contrast, RNA-seq analysis indicated that there were no substantial alterations in mRNA expression levels in the hippocampus of 3-months-old 5×FAD mice, compared to WT littermates. Strikingly, acRIP-seq revealed notable variations in ac4C modification on mRNAs, particularly those associated with synaptic structure and function, in the hippocampus of 3-months-old 5×FAD mice, compared with WT littermates. The ac4C modifications were found to be correlated with protein expression changes. Genes that are essential for synaptic function and cognition, including GRIN1, MAP2, and DNAJC6, exhibited reduced ac4C and protein levels in 3-months-old 5×FAD mice, without any corresponding changes in the mRNA levels, compared with WT littermates. Moreover, only a small part of dysregulated ac4C mRNAs identified in the 3-month-old 5×FAD mice were found in the 6-month-old 5×FAD mice. Conclusions Altogether these results identified abnormal ac4C modification of mRNAs that may contribute to the dysregulation of protein synthesis in the hippocampus from an early-stage mouse model of AD.
研究领域:
神经生物学
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分子生物学
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表观遗传学
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DOI
PubMed
高影响因子
Translational Neurodegeneration
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2025
·Vol. 14
·期 1
·1-27
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IF 15.2
Wang Jie,Ni Hong,Wang Yu,Wei Luyao,Ding Hanqing,Guo Zhongzhao,Pan Hao,Yu Ying,Luo Jia,Pan Weidong,Wang Deheng,Ke Zun-Ji
Background Alzheimer's disease (AD) is a neurodegenerative disease with major symptoms including memory and learning deficits. Neuroinflammation associated with reactive microglia promotes AD progression. These reactive microglia secrete prostaglandins, which are synthesized through the enzymatic activity of cyclooxygenase (COX)-1 and COX-2. Here, we aimed to elucidate the specific mechanisms of COX1 in AD pathogenesis and its interactions with neuroinflammatory processes.Methods We conducted backcrossing between COX-1 knockout (KO) and 5 × FAD mice to evaluate the effect of COX-1 deficiency on neuroinflammation. In addition, single-cell sequencing and microarray datasets from public databases and ingenuity pathway analysis in vitro were employed to explore gene expression profiles in the brains of AD mice.Results We identified a significant upregulation of COX-1 in 5 × FAD mice, with expression specifically localized to microglia in an age-dependent manner. Additionally, COX-1 KO alleviated neuroinflammation and accumulation of Aβ plaques, subsequently improving cognitive behavior in 5 × FAD mice. Moreover, microglia exhibited an amoeboid morphology in 5 × FAD mice, whereas in age-matched 5 × FAD/COX-1 KO mice, microglia had a ramified appearance. Additionally, our study demonstrated a pharmacological approach that inhibits the prostaglandin E2 (PGE2)/EP2 receptors via inhibition of the cAMP-PKA-NFκB-p65 pathway and NLRP3 inflammasome activation, producing similar beneficial effects as observed in COX-1 KO mice.Conclusion Our findings indicate that targeting the COX-1/PGE2/EP2 signaling pathway may alleviate neuroinflammation and impede AD progression. Moreover, the EP2 receptor presents a promising pharmacological target for mitigating the pathological effects associated with COX-1 activity in AD patients.
研究领域:
神经科学
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分子生物学
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药理学
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免疫学
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遗传学
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神经退行性疾病
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DOI
PubMed
高影响因子
ACS Synthetic Biology
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2026
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IF 4.5
Yunlong Wu,Yuhao Sun,Huan Miao,Xiaole Xia,Ling Gao
The biosynthesis of flavin adenine dinucleotide (FAD), an essential redox coenzyme, is limited by inefficient substrate utilization and low catalytic activity. To address this, an “Uptake-Catalysis” system was constructed in Bacillus subtilis for FAD synthesis from riboflavin (RF). First, the Streptomyces davawensis transporter (RibM) was coexpressed with native FAD synthase (BsFADs) at a 2:1 ratio, which increased RF consumption to 51%. Subsequently, the isothermal compressibility perturbation engineering (ICPE) strategy was adopted, generating the optimum mutant BsFADsP61W+R202Y, which increased catalytic efficiency (kcat/Km) toward RF and FMN by 83% and 73%, respectively. Mechanistic analysis revealed that this improvement was due to a 39% shortening of RF substrate channel. Finally, FAD titer peaked at 563 mg/L with a conversion rate (cr-FAD) of 92% in shaking-flask fermentation optimization. This work achieved a 225-fold increase in FAD production, validating the effectiveness of the uptake-catalysis synergistic strategy, thereby establishing an efficient and scalable synthesis paradigm.
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PubMed
高影响因子
Advanced Science
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2025
·
IF 14.1
Yue Sun,Dan Liu,Yu Liang,Xue Yang,Xinxin Liu,Baofeng Zhao,Zhen Liang,Yukui Zhang,Lihua Zhang
Spatially resolved characterization of proteoforms has substantial potential to significantly advance the understanding of physiological and disease mechanisms. However, challenges remain regarding throughput and coverage. A robust method is developed for high-throughput proteoform imaging (HTPi) by combining matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI MSI) and region-specific top-down proteomic analysis. MALDI MSI enables the imaging of proteoforms on tissue sections at a rate of 7 h cm −2 (100-µm spatial resolution), and the identification sensitivity of the proteoforms is improved by narrow-bore monolithic columns with low adsorption, yielding 366 annotated proteoform images from the mouse brain. The obtained proteoform images reveals differential expression of individual proteoforms across the brain regions, and distinct spatial distribution patterns of various proteoforms generated from a single gene. Given its ability to visualize proteoform, HTPi is further applied to explore spatial pathological changes associated with Alzheimer's disease (AD) in 5 × FAD mice. 158 annotated proteoform images are obtained in hippocampal regions at 50-µm spatial resolution, illuminating 14 differential proteoforms in the subiculum region and highlighting their significant associations with amyloid-β pathology in AD. The results highlight the power of HTPi in unraveling the intricate molecular landscape of brain tissues and its potential in elucidating disease mechanisms.
研究领域:
生物化学
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神经科学
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病理学
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2026
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IF 11.8
Hong Yang,Chen Liu,Yuzhen Wang,Zhen Wang,Yidan Zhang,Yuhua Lai,Lijie Zhang,Qiong Wu,Jinhua Liu
Existing multiplexed salivary diagnostics struggle to achieve a delicate balance between high sensitivity, low cost, and portability. This balance is essential for practical at-home diabetes monitoring. To address this challenge, we developed an integrated smartphone-based 3D paper sensor for multiplexed salivary metabolic monitoring featuring in situ -formed supraparticle zinc-coordination assemblies (SP-ZnCA) on wax-printed microzones. This design combines ratiometric fluorescence-using a DABSA probe with large Stokes shift and a multi-enzyme cascade (glucose oxidase, uricase, and cholesterol oxidase-horseradish peroxidase) for simultaneous quantification of salivary glucose, uric acid, and cholesterol with ultra-low detection limits (0.83-1.67 μM). We developed a custom modular micro fluorescence analysis device (μ-FAD) equipped with a 254 nm UV-LED, a 3D-printed dark chamber, and smartphone-based imaging. Through an optimized coaxial optical pathway, the μ-FAD enables rapid on-site testing within 8 minutes. Remarkably, the system reduces equipment costs to just 1/200 of those of conventional fluorometers. Clinical validation using 18 human saliva samples demonstrated high accuracy (R 2 > 0.97) relative to ELISA, at a mere $0.05 per test. By harmonizing material innovation, enzymatic signal amplification, and accessible diagnostics, our work successfully resolves the sensitivity-cost-portability trade-off and offers a scalable, affordable platform for decentralized diabetes management.
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PubMed
高影响因子
ELECTROCHIMICA ACTA
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2016
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IF 4.803
Chuantao Hou,Qiaolin Lang,Aihua Liu
Redox polymer hydrogels have been proved to be very promising for biofuel cells design and electrochemical biosensing. Here we report that a facile hydrogel bioanode based on electron-withdrawing nitro-group tailored 1,4-naphthoquinone (1,4-NQ)-appended redox polymer and flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH) can dramatically improve the catalytic performance of glucose oxidation , for which a high current density of 1.97 ± 0.06 mA cm −2 and low onset potential of −0.13 V (vs. Ag/AgCl) can be achieved in 50 mM glucose solution without using any nanomaterials as electrode supports. Further, both electron-donating groups modified 1,4-NQs and electron-withdrawing nitro-group 1,4-NQs were synthesized to investigate their electrochemical properties. It was found that the enhanced catalytic performance can be mainly contributing from the nitro group-modified 1,4-NQs appended redox polymer, which shifted the formal potential in the positive direction and provided sufficient electrochemical driving force for glucose oxidation. Finally, this hydrogel bioanode was combined with a bilirubin oxidase based biocathode to construct a single-compartment glucose/O 2 biofuel cell with a high power output of 0.28 ± 0.03 mW cm −2 and open circuit potential of 0.69 V. Moreover, the O 2 -insensitive FAD-GDH could improve the cell stability obviously compared with glucose oxidase based cell. This study provides an alternative route to develop glucose biosensor or biofuel cell for directly harvesting energy from glucose.
研究领域:
生物化学
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电化学
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能源材料
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产品关联
DOI
高影响因子
INTERNATIONAL IMMUNOPHARMACOLOGY
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2025
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IF 4.7
Rihong Luo,Yuhan Kang,He Ma,Zhenqiang Zhang,Christian Hölscher,Li Hao,Zijuan Zhang
To date, no therapeutic drugs available on the market can effectively reverse the progression of Alzheimer's disease (AD). Although Glucagon-like peptide-1 (GLP-1) receptor agonists (RAs) and Cholecystokinin (CCK) RAs have shown some promise in AD research, little is known about the neuroprotective effects of a novel dual CCK/GLP-1 RA in AD. This study sought to examine the effects of the novel dual CCK/GLP-1 RA on cognitive performance in an AD mouse model and to explore the associated mechanisms. Our findings indicate that dual CCK/GLP-1 RA improved cognitive deficits, reduced amyloid-beta (Aβ) accumulation, and alleviated mitochondrial damage in 5 × FAD mice by inducing mitophagy. In an in vitro model of AD cells induced by Aβ, CCK/GLP-1 RA could exert neuroprotective effects by regulating PINK1/Parkin-mediated mitophagy. These data reveal for the first time that the new CCK/GLP-1 RA modulates mitophagy via PINK1/Parkin pathway and enhances cognitive function in the 5 × FAD animal model. Moreover, the performance of the CCK/GLP-1 RA in certain indicators was superior to that of GLP-1 analogue liraglutide, suggesting that it may represent a more promising therapeutic option for AD.
研究领域:
神经科学
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药理学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
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