INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2020
·
IF 5.162
Xiaoyi Qu,Bo Ding,Jing Li,Meng Liang,Liqin Du,Yutuo Wei,Ribo Huang,Hao Pang
A novel β-glucosidase gene was isolated from Pseudoalteromonas sp. GXQ-1 and heterologously expressed in Escherichia coli . The activity of the encoded enzyme, PABGL, toward p -nitrophenyl-β-D-glucopyranoside was increased 8.74-fold by the presence of 3 M NaCl relative to the absence of added NaCl. PABGL hydrolyzed a variety of soy isoflavone substrates. For the conversion of daidzin to daidzein, the production rate was 1.44 mM/h. The addition of NaCl enhanced the hydrolytic activity of PABGL toward daidzin and genistein; the maximum activation by NaCl was 3.48- and 6.79-fold, respectively. This is the first report of a halophilic β-glucosidase from Pseudoalteromonas spp., and represents the β-glucosidase with the highest multiple of activation by NaCl. PABGL exhibits strong potential for applications in food processing and industrial production.
研究领域:
生物化学
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酶学
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微生物学
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食品科学
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生物技术
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PubMed
高影响因子
TOXICOLOGY
·
2023
·
IF 4.5
Miao Zhang,Xiaowen Bi,Shuai Liu,Yu Liu,Qiyu Wang
Sodium p -perfluorous nonenoxybenzene sulfonate (OBS), an economical alternative to perfluorooctane sulfonate (PFOS) in multiple industrial fields, is widely detected in the environment. The toxicity of OBS has received increasing attention. Pituitary cells are components of the endocrine system and act as vital regulators of homeostatic endocrine balance. However, the effects of OBS on pituitary cells remain unknown. The present study explores the effects of OBS (0.5, 5, and 50 μM) on GH3 rat pituitary cells after treatment for 24, 48, and 72 h. We found that OBS significantly inhibited cell proliferation in GH3 cells with remarkable senescent phenotypes, including enhanced SA-β-gal activity and expression of senescence-associated secretory phenotype (SASP)-related genes, cell cycle arrest, and upregulation of the senescence-related proteins γ-H2A.X and Bcl-2. OBS caused significant cell cycle arrest of GH3 cells at the G1-phase and concomitantly downregulated the expression of some key proteins for the G1/S transition, including cyclin D1 and cyclin E1. Consistently, the phosphorylation of retinoblastoma (RB), which plays a central role in regulating the cell cycle, was prominently reduced after OBS exposure. Furthermore, OBS notably activated the p53-p21 signalling pathway in GH3 cells, as evidenced by increased p53 and p21 expressions, enhanced p53 phosphorylation, and augmented p53 nuclear import. To our knowledge, this study is the first to reveal that OBS triggers senescence in pituitary cells via the p53-p21-RB signalling pathway. Our study demonstrates a novel toxic effect of OBS in vitro, and provides new perspectives for understanding the potential toxicity of OBS.
研究领域:
毒理学
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细胞生物学
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内分泌学
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PubMed
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2024
·Vol. 108
·期 1
·1-13
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IF 3.9
Wu Zongzhan,Dou Wenyu,Yang Xiaolin,Niu Tengfei,Han Zhuzhen,Yang Li,Wang Rufeng,Wang Zhengtao
Abstract Oleanane-type ginsenosides are a class of compounds with remarkable pharmacological activities. However, the lack of effective preparation methods for specific rare ginsenosides has hindered the exploration of their pharmacological properties. In this study, a novel glycoside hydrolase Pl GH3 was cloned from Paenibacillus lactis 154 and heterologous expressed in Escherichia coli . Sequence analysis revealed that Pl GH3 consists of 749 amino acids with a molecular weight of 89.5 kDa, exhibiting the characteristic features of the glycoside hydrolase 3 family. The enzymatic characterization results of Pl GH3 showed that the optimal reaction pH and temperature was 8 and 50 °C by using p -nitrophenyl- β - d -glucopyranoside as a substrate, respectively. The K m and k cat values towards ginsenoside Ro were 79.59 ± 3.42 µM and 18.52 s −1 , respectively. Pl GH3 exhibits a highly specific activity on hydrolyzing the 28- O - β - d -glucopyranosyl ester bond of oleanane-type saponins. The mechanism of hydrolysis specificity was then presumably elucidated through molecular docking. Eventually, four kinds of rare oleanane-type ginsenosides (calenduloside E, pseudoginsenoside RP1, zingibroside R1, and tarasaponin VI) were successfully prepared by biotransforming total saponins extracted from Panax japonicus . This study contributes to understanding the mechanism of enzymatic hydrolysis of the GH3 family and provides a practical route for the preparation of rare oleanane-type ginsenosides through biotransformation. Key points • The glucose at C-28 in oleanane-type saponins can be directionally hydrolyzed. • Mechanisms to interpret PlGH3 substrate specificity by molecular docking. • Case of preparation of low-sugar alternative saponins by directed hydrolysis.
研究领域:
生物化学
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药理学
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微生物学
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PubMed
高影响因子
Theranostics
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2023
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IF 12.4
Lianxia Guo,Haobin Cen,Jiaxian Weng,Yiting He,Xiaocao Guo,Di He,Kaisheng Liu,Shuyi Duan,Jing Yang,Xiaojian Zhang,Zifei Qin,Yong Wan,Zhiyong Chen,Baojian Wu
Rationale: The role of circadian clock in pituitary tumorigenesis remains elusive. Here we investigate whether and how circadian clock modulates the development of pituitary adenomas. Methods and Results: We found altered expression of pituitary clock genes in patients with pituitary adenomas. In particular, PER2 is prominently upregulated. Further, jetlagged mice with PER2 upregulation have accelerated growth of GH3 xenograft tumor. Conversely, loss of Per2 protects mice against developing estrogen-induced pituitary adenoma. Similar antitumor effect is observed for SR8278, a chemical that can decrease pituitary PER2 expression. RNA-seq analysis suggests involvement of cell cycle disturbance in PER2 regulation of pituitary adenoma. Subsequent in vivo and cell-based experiments validate that PER2 induces pituitary expression of Ccnb2 , Cdc20 and Espl1 (three cell cycle genes) to facilitate cell cycle progression and inhibit apoptosis, thereby promoting pituitary tumorigenesis. Mechanistically, PER2 regulates the transcription of Ccnb2 , Cdc20 and Espl1 through enhancing the transcriptional activity of HIF-1α. HIF-1α trans-activates Ccnb2 , Cdc20 and Espl1 via direct binding to its specific response element in the gene promoters. Conclusion: PER2 integrates circadian disruption and pituitary tumorigenesis. These findings advance our understanding of crosstalk between circadian clock and pituitary adenomas and highlight the relevance of clock-based approaches in disease management.
研究领域:
肿瘤学
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内分泌学
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分子生物学
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昼夜节律
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PubMed
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
·
2019
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IF 3.553
Xuejia Shi,Jin Xu,Changning Lu,Zhenzhong Wang,Wei Xiao,Linguo Zhao
The magnetic particle Fe 3 O 4 -SiO 2 -NH 2 -Cellu-ZIF8 (FSNCZ8) was used to immobilize the high-temperature resistant GH3 β-glucosidase (Tpebgl3) from the Thermomotoga petrophila DSM 13995. The Tpebgl3 has great potential in the catalytic conversion of pharmaceutically active components. The magnetic carrier (FSNCZ8) which provided stronger adsorption capacity, magnetic and high-temperature stability than the previously discussed Fe 3 O 4 -NH 2 -Cellu-ZIF8 without tetraethyl silicate coated. The properties of FSNCZ8-Tpebgl3 (FSNCZ8-T) were as follows: The optimal temperature and pH were 90 °C and 5.5, respectively; the highest activity of FSNCZ8-T approached 2672 U/g; Fe 2+ enabled immobilized enzyme to increase its relative activity to 194%. The main characteristics of FSNCZ8-T, including thermal stability, pH stability, and glucose tolerance, were greatly enhanced by adding Fe 2+ , which was also superior to free enzymes; moreover, the residual activity of FSNCZ8-T was 74% of the initial activity at the end of 10 repeated cycles.
研究领域:
生物化学
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酶工程
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材料科学
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PubMed
高影响因子
ACS Sustainable Chemistry & Engineering
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2026
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IF 7.6
Huabiao Miao,Junlong Wu,Menghan Yang,Yuqi Lu,Chunjiao Xie,Sha Wen,Bo Xu,Zunxi Huang,Chengbo Zhang
Human milk oligosaccharides (HMOs) and chitin oligosaccharides possess unique nutritional and prebiotic properties, driving interest in their sustainable synthesis. The key enzyme for their production is β-N-acetylhexosaminidase. Herein, we characterized Nag1420 (accession: PQ041313), a novel GH3 family enzyme from Cellulosilyticum sp., which exhibits both hydrolytic and transglycosylating activities. Substrate specificity analysis revealed a higher activity toward the natural substrate chitobiose than the synthetic analog p-nitrophenyl N-acetyl-β-d-glucosaminide (pNP-GlcNAc), enabling complete hydrolysis to N-acetylglucosamine. Notably, Nag1420 synthesized Lacto-N-triose II (LNT II) via transglycosylation when β-lactose was present as an acceptor. Crucially, in a cascade reaction with Chitinase, Nag1420 directly converted chitin waste into the high-value LNT II. This study provides insights into the green and low-cost biosynthesis of high-value-added carbohydrates.
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DOI
高影响因子
SCIENCE OF THE TOTAL ENVIRONMENT
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2017
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IF 4.9
Mi Deng,Yongming Wu,Chao Xu,Yuanxiang Jin,Xiaoli He,Jinbao Wan,Xiaoling Yu,Hongmin Rao,Wenqing Tu
A Chinese perfluorooctane sulfonate (PFOS) substitute frequently detected in the environment, 6:2 chlorinated polyfluorinated ether sulfonate (F-53B), has a similar structure to PFOS and it is proposed to cause thyroid dysfunction. To further confirm this hypothesis, the effects of F-53B on the thyroid endocrine system and underlying mechanisms were investigated in vitro and in vivo using rat pituitary GH3 cells and developing zebrafish, respectively. In GH3 cells, F-53B enhanced cell proliferation in a dose-dependent manner, indicative of thyroid receptor agonistic activity. In zebrafish larvae, F-53B exposure induced significant developmental inhibition and increased thyroxine (T 4 ) but not 3,5,3′-triiodothyronine (T 3 ) levels accompanied by a decrease in thyroglobulin (TG) protein and transcript levels of most genes involved in the hypothalamic-pituitary-thyroid (HPT) axis. Interestingly, T 4 levels remained significantly increased while TG protein and gene transcription levels were markedly upregulated after depuration. Molecular docking studies revealed that F-53B binds to transthyretin (TTR) by forming hydrogen bonds with Lys123 and Lys115, thereby interfering with thyroid hormone homeostasis. Our collective in vitro , in vivo and in silico studies provide novel evidence that F-53B disrupts the thyroid endocrine system at environmentally relevant concentrations, which cannot be recovered after depuration. Given the persistence of F-53B in the environment, the long-term consequences of thyroid hormone disruption by this chemical warrant further investigation.
研究领域:
环境科学
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毒理学
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内分泌学
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分子生物学
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DOI
PubMed
高影响因子
Microorganisms
·
2025
·Vol. 13
·期 9
·2033
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IF 4.2
Yanmei Cai,Xinyu Li,Shuang Chen,Qichao Liu,Hongxiang Lu,Jiahui Xie,Wei Li,Guiying Chen
Microorganisms with chitin-degrading capabilities play a crucial role in the biological control of crop pests and diseases as well as in the treatment of organic waste. In this study, a chitin-degrading bacterium, designated L2-2, was isolated from the intestine ofOdorrana margaretaecollected in Mount Emei, Sichuan, China. Based on physiological and biochemical characteristics, 16S rRNA gene sequencing, and phylogenetic analysis of 31 conserved housekeeping genes in the whole genome, strain L2-2 was identified as a member of the genusRoseateles, namedRoseatelessp. L2-2. This strain is able to grow on agar medium with colloidal chitin as the sole carbon source and form clear hydrolysis zones. After optimizing fermentation conditions (including concentrations of nitrogen and carbon sources, culture time, and pH), the enzyme activity was increased to 3.46 U/mL, which was 24 times higher than the initial enzyme activity. Functional genome annotation showed that the strain contains genes encoding endochitinases of the GH18, GH23, and GH46 families, as well as genes encoding β-glucosidases of the GH1, GH2, GH3, and GH109 families, indicating its genetic basis for chitin-degrading potential. This study expands the diversity of known chitin-degrading bacteria and provides a promising microbial resource for the bioremediation of chitinous waste and sustainable pest control in agriculture.
研究领域:
微生物学
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环境生物技术
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分子生物学
·
应用微生物学
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生物修复
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农业微生物学
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DOI
PubMed
高影响因子
Journal of Advanced Research
·
2024
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IF 11.4
Qin Yuan,Wen Liu,Wei Hao,Yi Chen,Yaqin Xiao,Hongyi Li,Mingju Shui,Ding-Tao Wu,Shengpeng Wang
Introduction Over decades, the source-function relationships of bioactive polysaccharides have been progressively investigated, however, it is still unclear how a defined structure may conduce to the bioactivities of polysaccharides. Objectives To explore the structure–function relationship of fungus polysaccharides, we employed a dextran sulfate sodium (DSS)‐induced colitis mouse model to compare the anti-inflammatory activity of two fungus polysaccharides from Dictyophora indusiata (DIP) and Tremella fuciformis (TFP), which exhibit distinct glycosidic linkages. Methods The structures of DIP and TFP were characterized through molecular weight detection, molecular morphology analysis, methylation analysis, and NMR analysis. Subsequently, we employed a DSS-induced colitis model to assess the anti-inflammatory efficacy of DIP and TFP. The colitis symptoms, histological morphology, intestinal inflammatory cytokines, and the composition and function of gut microbiota before and after polysaccharides treatment in colitis mice were also investigated. Results DIP, l,3-β-D-glucan with 1,4-β and 1,6-β-D-Glcp as branched chains, exhibited superior therapeutic effect than that of TFP consisted of a linear 1,3-α-D-mannose backbone with D-xylose and L-fucose in the side chains. Both DIP and TFP relieved DSS-induced colitis in a gut microbiota-dependent manner. Furthermore, metagenomics showed that DIP and TFP could partially reverse the bacterial function in colitis mice. Glycoside Hydrolase 1 (GH1) and GH3 were identified as being involved in hydrolyzing the glucose linkages in DIP, while GH92 and GH29 were predicted to be active in cleaving the α-1,3-linked mannose linkages and the glycosidic bonds of L-fucose residues in TFP. Conclusion Our findings highlight the pivotal role of glycosidic linkages in anti-inflammatory activities of fungus polysaccharides and would promote the design and discovery of polysaccharides with designated activity to be used as functional foods and/or therapeutics.
研究领域:
药理学
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免疫学
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微生物学
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生物化学
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分子生物学
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DOI
PubMed
高影响因子
Journal of Environmental Sciences
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2024
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IF 5.9
Zheyuan Song,Zheng Ma,Xueshan Feng,Rui Huang,Qiong An,Yunrui Pan,Jing Chang,Bin Wan,Huili Wang,Jianzhong Li
Ethiprole is widely used as a second-generation phenyl pyrazole insecticide. Previous study indicated that ethiprole exhibited thyroid toxicity while two main metabolites (ethiprole sulfone (M1) and ethiprole sulfide (M2)) of ethiprole showed higher acute toxicity than ethiprole. Therefore, assessing the thyroid toxicity of its metabolites is crucial for safety assessment. In this study, the thyroid toxicity and underlying mechanisms of ethiprole and its metabolites were explored using in silico, in vitro , and in vivo assays, with the aim of conducting a comparative study on thyroid toxicity. Molecular docking analysis showed that ethiprole, M1 and M2 could bind with thyroid receptor isoforms and exhibited higher binding affinity compared to 3,3′,5-triiodothyronine (T3). GH3 cell proliferation assays revealed that ethiprole, M1 and M2 all served as thyroid hormone antagonists to hinder the T3-induced cell proliferation. Using the zebrafish model, we further investigated that exposure to ethiprole, M1, and M2 disrupted thyroid hormone levels and the transcriptional expressions of hypothalamus-pituitary-thyroid (HPT) axis-related genes. Ethiprole induced thyroid disrupting effects by binding with the thyroid receptor beta, M1 mainly through binding with the corticotropin releasing factor receptor-1, and M2 exposure firstly inhibited the thyroid peroxidase enzyme activity. M2 showed the highest developmental toxicity and thyroid disrupting effects, which significantly reducing hatching rates, increasing deformity rates, exhibiting the lowest lethal concentration 50 value and showing the most serious transcription inhibitory effects on the HPT axis. This study suggested the risk assessment of metabolites should be considered in assessing potential environmental risk of ethiprole.
研究领域:
毒理学
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药理学
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环境科学
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分子生物学
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DOI
高影响因子
SCIENCE OF THE TOTAL ENVIRONMENT
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2023
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IF 9.8
Chao Hu,Yeqing Xu,Mingmin Wang,Shixuan Cui,Hangjun Zhang,Liping Lu
Bisphenol analogues are widely used in industrial and daily-use consumer products having imperfect thyroid hormones (THs) structures. Widespread exposure interferes with thyroid-related health outcomes in human. The mechanisms of disruption on TH synthesis and subsequent thyroid dysfunction by different bisphenol analogues remain unclear. Here, we evaluated bisphenol-induced thyroid endocrine disruption in C57BL/6 mice at doses of 0.002, 0.02, 2, and 20 mg/kg body weight/day (BW/d) for five consecutive weeks. Administration of 20 mg/kg BW/d bisphenol S (BPS) and 2 mg/kg BW/d tetrabromobisphenol S (TBBPS) significantly increased serum thyrotropin (TSH) levels to 1.21-fold and 1.20-fold of control group, respectively, indicating that bisphenols induced thyroid dysfunction in mice. Height of the thyroid follicle epithelium significantly increased to 1.27-, 1.24-, 1.26-, and 1.36-fold compared to control group with BPA, BPS, TBBPA, and TBBPS at 20 mg/kg BW/d, respectively, indicating impairment of the thyroid gland structure, and TBBPS showed potent effect. Exposure to bisphenol analogues of 0.02 mg/kg BW/d downregulated the protein expression levels of thyrotropin receptor, the sodium/iodide symporter, thyroperoxidase. The TH-dependent effects were further determined using the T-Screen assay at 10 −11 M to 10 −5 M concentrations. Bisphenol analogues significantly decreased TH-dependent GH3 cell proliferation, indicating the antagonistic activity of bisphenol analogues. The gene responsible for THs synthesis of thyrotropin releasing hormone receptor and TSH were upregulated, but downregulation of thyroid receptor β was observed. Our results suggest that bisphenol analogues distinctly induce thyroid dysfunction via TH synthesis, implying adverse effect of bisphenol analogues on TH homeostasis and subsequent physiological processes.
研究领域:
内分泌学
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毒理学
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环境健康
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产品关联
DOI
PubMed
高影响因子
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