INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Chang Liu,Qi Yan,Xuying Ding,Meijun Zhao,Chen Chen,Qian Zheng,Huiying Yang,Yining Xie
Brain-derived neurotrophic factor (BDNF) is a neurotrophic protein that promotes neuronal survival, increases neurotransmitter synthesis, and has potential therapeutic effects in neurodegenerative and psychiatric diseases, but its drug development has been limited by the fact that recombinant proteins of BDNF are unstable and do not penetrate the blood-brain barrier (BBB). In this study, we fused a TAT membrane-penetrating peptide with BDNF to express a recombinant protein (TBDNF), which was then PEG-modified to P-TBDNF. Protein characterization showed that P-TBDNF significantly improved the stability of the recombinant protein and possessed the ability to penetrate the BBB, and in cellular experiments, P-TBDNF prevented MPTP-induced nerve cell oxidative stress damage, apoptosis and inflammatory response, and its mechanism of action was closely related to the activation of tyrosine kinase B (TrkB) receptor and inhibition of microglia activation. In animal experiments, P-TBDNF improved motor and cognitive deficits in MPTP mice and inhibited pathological changes in Parkinson's disease (PD). In conclusion, this paper is expected to reveal the mechanism of action of P-TBDNF in inhibiting neurotoxicity, provide a new way for treating PD, and lay the foundation for the future development of recombinant P-TBDNF.
研究领域:
神经科学
·
药理学
·
分子生物学
·
神经退行性疾病
✓ 已验证
产品关联
DOI
PubMed
高影响因子
PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
·
2024
·
IF 4.2
Tengfei Xiong,Mingyue Yu,Jiang Zhu,Kai Tian,Mei Li,Xinghui Qiu
The oriental tobacco budworm Helicoverpa assulta (Lepidoptera: Noctuidae) is a specialist pest that may cause serious damages to important crops such as chili pepper and tobacco. Various man-made insecticides have been applied to control the infestation of this pest. To understand how this pest copes with insecticides, it is required to identify key players involved in insecticide transformation. In this study, a P450 gene of CYP6B subfamily was identified in the oriental tobacco budworm, and its expression pattern was revealed . Moreover, the activities of HassCYP6B6 against 12 insecticides were explored using recombinant enzymes produced in the facile Escherichia coli . Data from metabolic experiments showed that HassCYP6B6 was able to metabolize conventional insecticides including organophosporates (diazinon, malathion, phoxim), carbamate propoxur, and pyrethroid esfenvalerate, while no significant metabolism was observed towards new-type pesticides such as neonicotinoids (acetamiprid, imidacloprid), diamides (chlorantraniliprole, cyantraniliprole), macrocyclic lactone (emamectin benzoate, ivermectin), and metaflumizone. Structures of metabolites were proposed based on mass spectrometry analyses. The results demonstrate that HassCYP6B6 plays important roles in the transformation of multiple insecticides via substrate-dependent catalytic mechanisms including dehydrogenation, hydroxylation and oxidative desulfurization. The findings have important applied implications for the usage of insecticides.
研究领域:
昆虫学
·
分子生物学
·
毒理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
PEST MANAGEMENT SCIENCE
·
2024
·
IF 3.8
Xinyi Wang,Wenhong Li,Xiang Yang,Mingwei Yang,Yucheng Gu,Zhao Du,Jingyi Yang,Mingxia Wen,Yoonseong Park,Chunyang Huang,Yueping He
BACKGROUND Widespread resistance of insect pests to insecticides and transgenic crops in the field is a significant challenge for sustainable agriculture, and calls for the development of novel alternative strategies to control insect pests. One potential resource for the discovery of novel insecticidal molecules is natural toxins, particularly those derived from the venoms of insect predators. RESULTS In this study, we identified three insecticidal proteinaceous toxins from the venom glands (VGs) of the predatory stink bug, Arma custos (Hemiptera: Asopinae). Transcriptomic analysis of A. custos VGs revealed 151 potentially secreted VG-rich venom proteins. Three VG-rich venom proteins (designated AcVP1 ~ 3) were produced by overexpression in Escherichia coli . Injection of the recombinant proteins into tobacco cutworm ( Spodoptera litura ) larvae showed that all of the three recombinant proteins caused paralysis, liquefaction and death. Injection of recombinant proteins into rice brown planthopper ( Nilaparvata lugens ) nymphs showed higher insecticidal activities, among which a trypsin (AcVP2) caused 100% mortality postinjection at 1.27 pmol mg −1 body weight. CONCLUSION A natural toolkit for the discovery of insecticidal toxins from predatory insects has been revealed by the present study. © 2024 Society of Chemical Industry.
研究领域:
昆虫学
·
分子生物学
·
农业科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Biomolecules
·
2024
·Vol. 14
·期 12
·1553
·
IF 4.8
Zifu Ni,Linshang Zhang,Azhen Nie,Huan Wang,Xiaoling Wu
As the only naturally occurring β-amino acid, β-alanine has important application prospects in many fields. Driven by the huge demand, biosynthesis is becoming more and more popular as a potential alternative to the chemical synthesis of β-alanine. Although the direct pathway from L-aspartic acid to β-alanine, catalyzed by L-aspartic acid-α-decarboxylase (PanD), is ideal for β-alanine synthesis, it is hindered by the high cost of the substrate and limited economic viability. In this work, a cell-free dual enzyme cascade system based on methylaspartate lyase (EcMAL) and panD was constructed to safely and efficiently synthesize β-alanine using fumarate as a substrate. Taking the previously engineered EcMAL as the target, CgPanD was finally screened as the best candidate through gene mining, sequence alignment, and enzyme property analysis. Finally, under the optimal conditions of 35 °C, pH 8.0, and EcMAL: CgPanD concentration ratio of 1:5, the yield of β-alanine reached 80% theoretical yield within 120 min. This study provides a potential strategy for the biosynthesis of β-alanine, paving the way for future industrial-scale production.
研究领域:
生物化学
·
合成生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Foods
·
2024
·Vol. 13
·期 11
·1727
·
IF 4.7
Chengyu Lu,Ziwei Chen,Yuvaraj Ravikumar,Guoyan Zhang,Xinrui Tang,Yufei Zhang,Mei Zhao,Wenjing Sun,Xianghui Qi
L-Arabinose isomerase (L-AI) has been commonly used as an efficient biocatalyst to produce D-tagatose via the isomerization of D-galactose. However, it remains a significant challenge to efficiently synthesize D-tagatose using the native (wild type) L-AI at an industrial scale. Hence, it is extremely urgent to redesign L-AI to improve its catalytic efficiency towards D-galactose, and herein a structure-based molecular modification ofLactobacillus plantarumCY6 L-AI (LpAI) was performed. Among the engineered LpAI, both F118M and F279I mutants showed an increased D-galactose isomerization activity. Particularly, the specific activity of double mutant F118M/F279I towards D-galactose was increased by 210.1% compared to that of the wild type LpAI (WT). Besides the catalytic activity, the substrate preference of F118M/F279I was also largely changed from L-arabinose to D-galactose. In the enzymatic production of D-tagatose, the yield and conversion ratio of F118M/F279I were increased by 81.2% and 79.6%, respectively, compared to that of WT. Furthermore, the D-tagatose production of whole cells expressing F118M/F279I displayed about 2-fold higher than that of WT cell. These results revealed that the designed site-directed mutagenesis is useful for improving the catalytic efficiency of LpAI towards D-galactose.
研究领域:
生物化学
·
食品科学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
PROCESS BIOCHEMISTRY
·
2024
·
IF 3.7
Long Zhang,Jinna Cui,Wei Liu,Zhanying Liu
Various protein secretion systems of gram-negative Escherichia coli ( E. coli ) are well known, but the secretion pathway of endoglucanase (BcsZ) is unknown. To further elucidate the secretion mechanism, we analyzed the transcriptome data of five ruminal cellulolytic E. coli strains. Gene Ontology (GO) enrichment analysis revealed that the differentially expressed genes (DEGs) upregulated in all five strains were significantly enriched in the membrane. Nine DEGs related to membrane transporters were overexpressed in E. coli JBZ-DH5α (overexpressing BcsZ in the genome of E. coli DH5α). The results revealed that the overexpression of the b4332 ( yjiJ ), b2142 ( yohK ), and b2377 ( yfdY ) genes significantly reduced the growth and BcsZ concentration of the strain. The overexpression of the genes b3533 ( bcsA ), b3009 ( yghB ), and b3522 ( yhjD ) did not significantly affect strain growth or the BcsZ concentration. The overexpression of the b1859 ( znuB ), b0458 ( ylaC ), and b1195 ( ymgE ) genes increased the secretion of BcsZ but had no significant effect on strain growth. In conclusion, the yjiJ , yohK , yfdY , znuB , ylaC , and ymgE genes are involved in the secretion of E. coli BcsZ. These findings provide insight into the secretion mechanism of E. coli BcsZ.
研究领域:
微生物学
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
FOOD RESEARCH INTERNATIONAL
·
2024
·
IF 7
Chenyu Song,Wei Xu,Cuie Guang,Ting Xue,Wanmeng Mu
Patulin (PAT) is a mycotoxin commonly found in fruits and vegetables, prompting the need for effective removal and detoxification methods, which have garnered significant research attention in recent years. Among these methods, the utilization of microbial-derived enzymes stands out due to their mild operating conditions, specificity in targeted functional groups, and the production of non-toxic by-products, making it a preferred degradation approach. In this study, a novel PAT-degrading enzyme derived from Cyberlindnera fabianii ( Cyfa -SDR) was identified, demonstrating its highest catalytic activity at pH 7.0 and 80 °C against PAT. This temperature tolerance level represents the highest reported for PAT-degrading enzymes to date. The enzyme was further characterized as a short-chain dehydrogenase through analysis of its amino acid composition, conserved GXXXGXG motif, and dependency on NADPH. Moreover, the study evaluated the efficiency of PAT degradation by Cyfa -SDR at varying substrate and enzyme concentrations, surpassing the performance of other PAT-degrading enzymes, thus highlighting its substantial potential for the biological control of PAT. In conclusion, the enzymatic treatment using the PAT-degrading enzyme Cyfa -SDR presents a viable and promising solution for enhancing the quality and safety of fruit juice.
研究领域:
食品科学
·
微生物学
·
酶学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Jingjing Zhang,Jing Huang,Yufan Yang,Daxin Zhao,Cong Wang,Peng Zhong,Jianhua Jia,Weifan Dang,Qiwei Lu,Chengyu Zhang,Xiaohui Yan
Icaritin (ICT), a compound with diverse biological activities derived from Epimedium folium, is typically present in low concentrations in EFs. However, the abundant glycosyl-modified ICT compounds facilitate its transformation into ICT. Current biocatalytic production faces challenges, including low conversion rates and limited enzyme activity. This study developed a one-pot enzymatic cascade strategy for directly biotransform crude extracts of Epimedium folium (EEF) to produce ICT. The feasibility of catalyzing different ICT-related compounds in EEF was validated through molecular docking and substrate reactions. The selected glycosidase exhibited simultaneous activities as a glucosidase, xylosidase, and α-1,6-rhamnosidase, with the rhamnosidase showing outer-rhamnosidic activity and weak glucosidase activity. By using EFs as the substrate and employing whole-cells ( Escherichia coli ) containing LacS and BtRha proteins for synergistic catalysis, icariin can be efficiently synthesized within 6 h, achieving a conversion rate of 100 %. The enzymatic cascade for ICT production from crude extracts was elucidated by analyzing catalytic intermediates via HPLC. Compared to strategies using single or traditional multi-enzyme applications, this method shows advantages of ease to operation, high efficiency, and large production yield performance. This method has the potential to become an eco-friendly catalytic strategy for the large-scale production of icaritin.
研究领域:
生物技术
·
生物化学
·
生药学
·
酶学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
STAR Protocols
·
2024
·
IF 1.3
Yuanyan Wei,Shuting Geng,Qihang Chen,Jiayue Chen,Zhijun Fan,Yi Li,Yu Si,Yuerong Yang,Ziwei Liang,Jianhai Jiang
Summary CD133 is widely used as a marker to isolate cancer stem cells (CSCs). However, the structural ambiguity of N-glycan of CD133 limits its application in the isolation of CSCs. Here, we present a protocol to sort CSCs from tumor samples by combining CD133 with α-1,2-high-mannose type glycan chains. We describe steps for purifying and biotinylating cyanovirin-N (CVN) proteins and sorting tumor cells and CD133 + α-1,2-Man + (α-1,2-linked mannose + ) cells to provide more specific markers for CSCs. For complete details on the use and execution of this protocol, please refer to Wei et al. 1
研究领域:
肿瘤学
·
细胞生物学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Bingjie Xu,Dan Yu,Chenlu Xu,Yujie Gao,Hengqiu Sun,Lei Liu,Yang Yang,Dongming Qi,Jindan Wu
Innovative antibacterial therapies using nanomaterials, such as photothermal (PTT) and photodynamic (PDT) treatments, have been developed for treating wound infections. However, creating secure wound dressings with these therapies faces challenges. The primary focus of this study is to prepare an antibacterial nanofiber dressing that effectively incorporates stable loads of functional nanoparticles and demonstrates an efficient synergistic effect between PTT and PDT. Herein, a composite nanofiber mat was fabricated, integrating spherical molybdenum disulfide (MoS 2 ) nanoparticles. MoS 2 was deposited onto polylactic acid (PLA) nanofiber mats using vacuum filtration, which was further stabilized by sodium carboxymethyl cellulose (CMC) adhesion and glutaraldehyde (GA) cross-linking. The composite nanofibers demonstrated synergistic antibacterial effects under NIR light irradiation, and the underlying mechanism was explored. They induce bacterial membrane permeability, protein leakage, and intracellular reactive oxygen species (ROS) elevation, ultimately leading to >95 % antibacterial activity against Staphylococcus aureus ( S. aureus ) and Escherichia coli ( E. coli ), which is higher than that of single thermotherapy (almost no antibacterial activity) or ROS therapy (about 80 %). In addition, the composite nanofiber mats exhibited promotion effects on infected wound healing in vivo. This study demonstrates the great prospects of composite nanofiber dressings in clinical treatment of bacterial-infected wounds.
研究领域:
生物医学工程
·
纳米医学
·
感染与免疫
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Synthetic Biology
·
2024
·
IF 3.7
Jialing Wang,Jie Chen,Kemin Lv,Zhen Gao,Jiahuang Li,Bin Wu,Bingfang He,Gerhard Schenk
Processive endoglucanases have generated significant interest due to their bifunctionality in the degradation of cellulose and low product inhibition. However, enhancing their catalytic efficiency through engineering remains a formidable challenge. To address this bottleneck, our engineering efforts targeted loop regions located in the substrate channel of processive endoglucanase EG5C-1. Guided by a comparative analysis of characteristic structural features of the substrate channels in cellobiohydrolase, endoglucanase, and processive endoglucanase, a highly active triple mutant CM6 (N105H/T205S/D233L) was generated that had a 5.1- and 4.7-fold increase in catalytic efficiency toward soluble substrate carboxymethyl cellulose-Na and insoluble substrate phosphoric acid-swollen cellulose (PASC), compared with wild-type EG5C-1. Furthermore, this mutant exhibited greater processivity compared to EG5C-1. Molecular dynamics simulations unveiled that the mutations in the loop regions reshaped the substrate channel, leading to a deeper cleft, resembling the closed channel configuration of cellobiohydrolases. The increased compactness of the substrate channel induced changes in the substrate binding mode and substrate deformation, thereby enhancing both binding affinity and catalytic efficiency. Moreover, metadynamics simulations demonstrated that the processive velocity of cellulose chain through the binding channel in mutant CM6 surpassed that observed in EG5C-1.
研究领域:
生物化学
·
酶工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF BIOTECHNOLOGY
·
2025
·
IF 3.9
Yuwen Wei,Fulong Li,Yukun Zheng,Youxiang Liang,Yan Du,Huimin Yu
Transaminases have important applications in the synthesis of drug intermediates such as chiral amines. However, natural transaminases exhibit suboptimal thermal stability, limiting their further applications. Building upon an Rhodobacter sp. -derived ( R )-selective transaminase ( Rb TA), we report a dual-region coupling engineering approach to improve thermostability of Rb TA by strengthening the core hydrogen-bond networks and rigidifying the flexible surface loop. Through single strategy, we identified 4 thermostability improved single mutations, among which I249Q demonstrated the most substantial improvement, achieving a 18-fold increase in half-life ( t 1 / 2 40 ) and a 11.2 ℃ increase in T 50 10 . Then in strategic coupling, the synergistic effect of dual-region modification was observed in both thermal stability and activity enhancement, as mutant with the best high-temperature catalytic performance, R136P/F228Y, had its T 50 10 improved by 7.1℃ and exhibited a 4.2-fold increase in k cat / K m towards ( R )-3-amino-1-butanol. Finally, R136P/F228Y achieved a 20.5 % improvement in conversion over WT in an analytical-scale synthesis in 72 h at a 5 ℃ elevated catalytic temperature. Molecular dynamics simulations demonstrated that the synergy of the formation of new hydrogen bonds and decrease in flexibility accounted for the thermostability improvements. This study provides guidance for enhancing thermostability of similar fold-type enzymes without impairing enzymatic activity in an efficient manner.
研究领域:
生物化学
·
蛋白质工程
✓ 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）