PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
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2024
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IF 4.2
Pan-Pan Guo,Xi-Bin Yang,Hong Yang,Cao Zhou,Gui-Yun Long,Dao-Chao Jin
β -N-Acetylglucosaminidases and/or β - N -acetylhexosaminidases (NAGs / Hexes) are crucial exonucleases, playing a crucial role in the insect molting process. SfHex3 and SfHex4 contain conserved catalytic domains of GH20 and GH20b, clustered into NAG2 and NAG1 group, respectively. SfHex3 and SfHex4 were mainly highly expressed in the 4th–5th instar nymphs, as well as in the integument and ovary. The expression level of SfHex3 gradually decreased in male and female adults, and SfHex4 on the first day of female was significantly higher than that on the first day of male. In addition, RNA interference (RNAi) results demonstrated that the downregulation of SfHex3 and SfHex4 expression in 5th-instar nymphs resulted in failed molting, and a high mortality. Furthermore, after RNAi with SfHex3 and SfHex4 , the transcript levels on key genes of the chitin metabolism pathway ( SfCHS1 , SfCHS1a , SfCHS1b , SfTRE1 , SfTRE2 , SfCht5 , and SfCht7 ) were significantly decreased compared to the control group. Meanwhile the expression levels of SfHex3 and SfHex4 were up-regulated after 6 h and 12 h of 20E treatment. And the transcription levels of SfHex3 and SfHex4 were significantly inhibited at nitenpyram LC 20 , LC 50 , and LC 90 after 96 h of treatment, in 3rd nymphs of Sogatella furcifera . In conclusion, SfHex3 and SfHex4 play important roles in the nymphal development of S. furcifera , contributing to the molting process from nymph to adult. This study not only enhances our understanding of the nitenpyram in pest control, but also provides a foundation for the development of new control strategies using RNAi to targeting SfHex3 and SfHex4 .
研究领域:
昆虫学
·
分子生物学
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DOI
高影响因子
ANAIS DA ACADEMIA BRASILEIRA DE CIENCIAS
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2018
·Vol. 90
·期 2
·1503-1519
·
IF 0.956
JIN CUILI,YU ZHAOWEI,PENG SHUYA,FENG KE,ZHANG LIKUI,ZHOU XIAOJIAN
Exopolysaccharide (EPS) of two benthic diatoms, Amphora sp. and Stauroneis sp., with different biofilm formation abilities were investigated. The ratio of suspension-cells/biofilm-cells was employed to indicate the diatom biofilm formation abilities. The soluble EPS from the supernatant of whole culture, tightly bound EPS from floating cells, loosely and tightly bound EPS from biofilm cells were fractionated as SL-EPS, F-TB-EPS, BF-LB-EPS and BF-TB-EPS, respectively. The analysis for productions and monosaccharide compositions indicated that EPS from two diatoms were different in terms of the productions, distributions, and monomer compositions. Amphora sp. produced more (1.5-fold) total exopolysaccharides, but less (<0.4-fold) BF-TB-EPS than Stauroneis sp. The monosaccharides of the EPS from Amphora sp. were more diverse than those of Stauroneis sp., with 13 and 10 monomers, respectively. Neutral sugars, Glc, Xyl and Man, were abundant in Stauroneis sp., while Gal, Glc and Xyl were rich in Amphora sp. Uronic acid and hexosamine were present in all fractions of two diatoms, especially Glc-A being the most abundant monomer in SL-EPS of Amphora sp. It was proposed that the high content of uronic acid (especially Glc-A) might be crucial for the strong biofilm formation abilities of Amphora sp.
研究领域:
微生物学
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海洋生物学
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生物技术
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产品关联
DOI
PubMed
高影响因子
Frontiers in Molecular Biosciences
·
2021
·
IF 5.246
Qu Tianle,Zhang Chunyue,Qin Zhen,Fan Liqiang,Jiang Lihua,Zhao Liming
Aminooligosaccharides possess various biological activities and can exploit wide applications in food, pharmaceutical and cosmetic industries. Commercial aminooligosaccharides are often prepared by the hydrolysis of chitin and chitosan. In this study, a novel GH family 20 β-N-acetylhexosaminidases gene named AoNagase was cloned from Aspergillus oryzae and expressed in Pichia pastoris. The purified AoNagase had maximal activity at pH 5.5 and 65°C. It exhibited good pH stability in the range of pH 6.0–7.5 and at temperatures below 50°C. AoNagase was capable of hydrolyzing not only colloidal chitosan (508.26 U/mg) but also chitin (29.78 U/mg). The kinetic parameters (Km and Vmax) of AoNagase were 1.51 mM, 1106.02 U/mg for chitosan and 0.41 mM, 40.31 U/mg for colloidal chitin. To our knowledge, AoNagase is the first GH family 20 β-N-acetylhexosaminidase capable of hydrolyzing both chitosan and chitin. AoNagase is an endo-type β-N-acetylhexosaminidases and can potentially be used for the manufacturing of aminooligosaccharides.
研究领域:
生物化学
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酶学
·
生物技术
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产品关联
DOI
PubMed
高影响因子
PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
·
2024
·
IF 4.2
Ming Liu,Qianwen Wang,Bin Lai,Yan Chen,Rongchumu Ge,Shuo Yan,Chunya Bu
β- N -acetylhexosaminidases (HEXs) are key chitin-degrading enzymes in insects. Here, we identified TuHex1 and TuHex2 using insect orthologous genes by searching Tetranychus urticae genome and transcriptome database to investigate their roles in mite molting. TuHex1 and TuHex2 expression was induced by 20-hydroxyecdysone (20E), and inhibition of TuHex1 and TuHex2 expression by RNAi resulted in wrinkled cuticle or an inability to shed the old cuticle in nymphs, which may be due to a reduction in particle deposition in the exocuticle and lamellar structure in the endocuticle as revealed by scanning electron microscopy and transmission electron microscopy. The results suggest that the TuHex1 and TuHex2 genes play an essential role in the molting and developmental process of the mite. TuHex2 , with a mortality rate of 67.41 % in the leaf disc assay, was a potential RNAi target by oral feeding. Spraying of nanocarrier-delivered bacteria expressing ds TuHex2 at 500 ng/μL kept spider mites at a consistently low level throughout the 14 days and showed good mite control comparable to that of matrine. In addition, nanocarrier-delivered ds TuHex2 is safe for Neoseiulus californicus in our experiments, providing its potential for green mite management.
研究领域:
昆虫学
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分子生物学
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害虫管理
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF ASIA-PACIFIC ENTOMOLOGY
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2017
·
IF 1.046
Yunhua Zhang,Jinyun Ji,Xiao Zhao,Ruoheng Jin,Jianhong Li,Hu Wan
A previous study demonstrated that a β- N -acetyl- d -hexosaminidase ( NlHex4 ) from the brown planthopper could serve as a new potential target for biopesticides due to its requirement for molting, and lethal phenotypes were observed via RNA interference . However, the enzymatic properties of NlHex4 remain unclear. In the present study, NlHex4 was expressed, purified and characterized. Recombinant NlHex4 was functionally expressed in baculovirus-infected insect cells and purified with a molecular weight of approximately 70 kDa. Recombinant NlHex4 had an optimal pH and temperature of 6.0 and 60 °C, respectively. Moreover, recombinant NlHex4 exhibited thermal stability at 50 °C for at least 1 h. Furthermore, the activity of recombinant NlHex4 was inhibited by the metal ions Fe 2 + and Fe 3 + and the serine protease inhibitor phenylmethanesulfonyl fluoride. These results reveal the enzymatic properties of NlHex4 and provide a basis for future investigation of the interaction between HEX and chitin oligosaccharide .
研究领域:
昆虫学
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生物化学
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害虫管理
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产品关联
DOI
高影响因子
JOURNAL OF SEPARATION SCIENCE
·
2024
·Vol. 47
·期 1
·2300545
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IF 2.8
Chen Yang,Jing-Wen Zhang,Jin Qi,Bo-Yang Yu
Pseudoallergy is a typical and common adverse drug reaction to injections, especially in traditional Chinese medicine injections (TCMIs). At present, the evaluation methods for pseudoallergy include cell methods in vitro and animal methods in vivo. The mast cell evaluation method based on the β-hexosaminidase (β-Hex)-catalyzed substrate, 4-nitrophenyl-β-N-acetyl-D-glucosaminide (4-NPG), is an important method for the evaluation of drug-induced pseudoallergy, but it is prone to false positive results and has insufficient sensitivity. In this study, a novel β-Hex evaluation system with rat basophilic leukemia-2H3 cells based on high-performance liquid chromatography-fluorescence detection (HPLC-FLD) was established, which effectively increased the sensitivity and avoided false positive results. Cell viabilities were measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide assay. In addition, a method for the determination of histamine, which is another indicator in the development of pseudoallergy, was established to validate the above method. The results of this novel method indicated that two TCMIs (Shuxuening injection and Shenqi Fuzheng injection), which were considered to be pseudoallergenic using 4-NPG, were not pseudoallergenic. Overall, the novel β-Hex/HPLC-FLD evaluation system using Rat basophilic leukemia-2H3 cells established was effective and precise. It could be used for the evaluation of pseudoallergic reactions caused by TCMIs and other injections.
研究领域:
药理学
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免疫学
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分析化学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
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IF 8.5
Jiajun Yang,Siyu Hou,Yuhui Zhao,Zhaoyang Sun,Lilin Zhang,Yan Deng,Xiaoli Shang,Hanjie Yu,Zheng Li,Hongmei Li
The antidiabetic activity of the novel Buckwheat protein-derived peptide (Ala-Phe-Tyr-Arg-Trp, AFYRW) and its associated protein glycosylation have been verified. Our preliminary study demonstrates the potential of AFYRW as a therapeutic agent for diabetes, but the mechanism needs further investigation. Given the vital role of O-linked N -acetylglucosamine transferase (OGT) in diabetes mellitus and insulin resistance (IR), we focused on the underlying molecular mechanisms of them in ameliorating IR. We found AFYRW protects against hyperglycemia in diabetic mice and improves glucose metabolism in an IR cell model. Mechanistically, we demonstrated that AFYRW decreases glutamine-fructose-6-phosphate amidotransferase (GFAT) expression via X-box binding protein 1 (XBP1) in the hexosamine biosynthesis pathway (HBP), consequently decreasing OGT and stimulating the SIRT1/PGC1α pathway. Of note, the overlapping roles of increased SIRT1 and decreased OGT caused by AFYRW ameliorated IR. The data presented here show that AFYRW contributes to metabolism by directly controlling glucose homeostasis. Taken together, our study unveils that AFYRW protects against both insulin resistance and diabetes mellitus-induced hyperglycemia through OGT to regulate the SIRT1/PGC1α pathway, which provides a mechanistic basis for novel AFYRW to be a potential therapeutic agent.
研究领域:
内分泌学
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分子生物学
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糖尿病研究
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药理学
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产品关联
DOI
PubMed
高影响因子
PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
·
2017
·
IF 1.361
Xiaoyan Li,Chen Qi,Peilian Wei,Lei Huang,Jin Cai,Zhinan Xu
Uridine 5′-diphosphate N-acetylglucosamine (UDP-GlcNAc) is a natural UDP-monosaccharide donor for bacterial glycosyltransferases, while uridine 5′-diphosphate N-trifluoacetyl glucosamine (UDP-GlcNTFA) is its synthetic mimic. The chemoenzymatic synthesis of UDP-GlcNAc and UDP-GlcNTFA was attempted by three recombinant enzymes. Recombinant N-acetylhexosamine 1-kinase was used to produce GlcNAc/GlcNTFA-1-phosphate from GlcNAc/GlcNTFA. N-acetylglucosamine-1-phosphate uridyltransferase from Escherichia coli K12 MG1655 was used to produce UDP-GlcNAc/GlcNTFA from GlcNAc/GlcNTFA-1-phosphate. Inorganic pyrophosphatase from E. coli K12 MG1655 was used to hydrolyze pyrophosphate to accelerate the reaction. The above enzymes were expressed in E. coli BL21 (DE3) and purified, respectively, and finally mixed in one-pot bioreactor. The effects of reaction conditions on the production of UDP-GlcNAc and UDP-GlcNTFA were characterized. To avoid the substrate inhibition effect on the production of UDP-GlcNAc and UDP-GlcNTFA, the reaction was performed with fed batch of substrate. Under the optimized conditions, high production of UDP-GlcNAc (59.51 g/L) and UDP-GlcNTFA (46.54 g/L) were achieved in this three-enzyme one-pot system. The present work is promising to develop an efficient scalable process for the supply of UDP-monosaccharide donors for oligosaccharide synthesis.
研究领域:
生物化学
·
酶学
·
合成生物学
·
微生物学
✓ 已验证
DOI
PubMed
高影响因子
Frontiers in Immunology
·
2022
·
IF 8.786
Yang Yong-Shi,Cao Meng-Da,Wang An,Liu Qing-Mei,Zhu Dan-Xuan,Zou Ying,Ma Ling-Ling,Luo Min,Shao Yang,Xu Dian-Dou,Wei Ji-Fu,Sun Jin-Lyu
Background: Allergic respiratory diseases have increased dramatically due to air pollution over the past few decades. However, studies are limited on the effects of inorganic components and particulate matter with different particle sizes in smog on allergic diseases, and the possible molecular mechanism of inducing allergies has not been thoroughly studied.Methods: Four common mineral elements with different particle sizes in smog particles were selected, including Al2O3, TiO2, Fe2O3, and SiO2. We studied the relationship and molecular mechanism of smog particle composition, particle size, and allergic reactions using mast cells, immunoglobulin E (IgE)-mediated passive cutaneous anaphylaxis (PCA) model, and an ovalbumin (OVA)-induced asthmatic mouse model in vitro and in vivo, combined with transmission electron microscopy, scanning transmission X-ray microscopy analysis, and transcriptome sequencing.Results: Only 20 nm SiO2 particles significantly increased β-hexosaminidase release, based on dinitrophenol (DNP)-human serum albumin (HSA) stimulation, from IgE-sensitized mast cells, while other particles did not. Meanwhile, the PCA model showed that Evan’s blue extravasation in mice was increased after treatment with nano-SiO2 particles. Nano-SiO2 particles exposure in the asthmatic mouse model caused an enhancement of allergic airway inflammation as manifested by OVA-specific serum IgE, airway hyperresponsiveness, lung inflammation injury, mucous cell metaplasia, cytokine expression, mast cell activation, and histamine secretion, which were significantly increased. Nano-SiO2 particles exposure did not affect the expression of FcϵRI or the ability of mast cells to bind IgE but synergistically activated mast cells by enhancing the mitogen-activated protein kinase (MAPK) signaling pathway, especially the phosphorylation levels of the extracellular signal-regulated kinase (ERK)1/2. The ERK inhibitors showed a significant inhibitory effect in reducing β-hexosaminidase release.Conclusion: Our results indicated that nano-SiO2 particles stimulation might synergistically activate IgE-sensitized mast cells by enhancing the MAPK signaling pathway and that nano-SiO2 particles exposure could exacerbate allergic inflammation. Our experimental results provide useful information for preventing and treating allergic diseases.
研究领域:
免疫学
·
环境健康
·
分子生物学
·
过敏与哮喘
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）