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Mechanistic insights into the conversion of flavin adenine dinucleotide(FAD)to 8-formyl FAD in formate oxidase:a combined experimental and in-silico study

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Bioresources and Bioprocessing 2024年第1期11卷 909-921页

作者： Kai Wen Sirui Wang Yixin Sun Mengsong Wang Yingjiu Zhang Jingxuan Zhu Quanshun Li Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education School of Life SciencesJilin UniversityChangchun 130012China Center for Supramolecular Chemical Biology Jilin UniversityChangchun 130012China

Formate oxidase(FOx),which contains 8-formyl flavin adenine dinucleotide(FAD),exhibits a distinct advantage in utilizing ambient oxygen molecules for the oxidation of formic acid compared to other glucose-methanol-choline(GMC)oxidoreductase enzymes that contain only the standard FAD *** FOx-mediated conversion of FAD to 8-formyl FAD results in an approximate 10-fold increase in formate oxidase ***,the mechanistic details underlying the autocatalytic formation of 8-formyl FAD are still not well understood,which impedes further utilization of *** this study,we employ molecular dynamics simulation,QM/MM umbrella sampling simulation,enzyme activity assay,site-directed mutagenesis,and spectroscopic analysis to elucidate the oxidation mechanism of FAD to 8-formyl *** results reveal that a catalytic water molecule,rather than any catalytic amino acids,serves as a general base to deprotonate the C8 methyl group on FAD,thus facilitating the formation of a quinone-methide tautomer *** oxygen molecule subsequently oxidizes this intermediate,resulting in a C8 methyl hydroperoxide anion that is protonated and dissociated to form OHC-RP and OH−.During the oxidation of FAD to 8-formyl FAD,the energy barrier for the rate-limiting step is calculated to be 22.8 kcal/mol,which corresponds to the required 14-hour transformation time observed ***,the elucidated oxidation mechanism reveals that the autocatalytic formation of 8-formyl FAD depends on the proximal arginine and serine residues,R87 and S94,*** activity assay validates that the mutation of R87 to lysine reduces the kcat value to 75%of the wild-type,while the mutation to histidine results in a complete loss of ***,the mutant S94I also leads to the deactivation of *** dependency arises because the nucleophilic OH−group and the quinone-methide tautomer intermediate are stabilized through the noncovalent interaction provided by R8

关键词： Formate oxidase 8-Formyl flavin adenine dinucleotide Oxidative maturation molecular dynamics simulation QM/MM umbrella sampling simulation

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Light/pH dual-responsive magnetic metal-organic frameworks composites for phosphorylated peptide enrichment

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Chinese Chemical Letters 2024年第7期35卷 222-226页

作者： Rui Wang He Qi Haijiao Zheng Qiong Jia College of Chemistry Jilin UniversityChangchun 130012China Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education School of Life SciencesJilin UniversityChangchun 130012China

Metal-organic frameworks(MOFs)combined with specific ligands are highly adaptable smart materials that can respond to external and physiological *** this study,we introduced a pyridinyl zwitterionic ligand with light/pH dual response into magnetic MOF composite(Fe_(3)O_(4)@ZW-MOF)for enrichment of phosphorylated peptides for the first *** introduction of the developed ligand gives MOF material dual response *** stimulation affects the generation/disappearance of free radicals of the pyridine derivative,resulting in a change in the charge gradient of the zwitterion,and zwitterion can also regulate the p H of the solution by adding acid or ***,the reversible capture and release of phosphorylated peptides can be easily achieved by adjusting light and *** established phosphorylated peptide enrichment platform exhibits high sensitivity(detection limit of 1 fmol),high selectivity(β-casein:BSA,1:1000),and good reusability(7 cycles).In addition,the method was applied to the enrichment of phosphorylated peptides in complex systems(non-fat milk and human serum),demonstrating the feasibility of this method for phosphoproteom *** conclusion,the synthesized Fe_(3)O_(4)@ZW-MOF is a promising MOF material,which provides the possibility to advance the application of responsive MOFs materials in proteomics.

关键词： Light pH Dual-response Metal-organic frameworks Phosphorylated peptide

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Self-assembled multiepitope nanovaccine based on NoV P particles induces effective and lasting protection against H3N2 influenza virus

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Nano Research 2023年第5期16卷 7337-7346页

作者： Jiaojiao Nie Qingyu Wang Shenghui Jin Xin Yao Lipeng Xu Yaotian Chang Fan Ding Zeyu Li Lulu Sun Yuhua Shi Yaming Shan National Engineering Laboratory for AIDS Vaccine School of Life SciencesJilin UniversityJilin 130012China Key Laboratory for Molecular Enzymology and Engineering The Ministry of EducationSchool of Life SciencesJilin UniversityJilin 130012China

Current seasonal influenza vaccines confer only limited coverage of virus strains due to the frequent genetic and antigenic variability of influenza virus(IV).Epitope vaccines that accurately target conserved domains provide a promising approach to increase the breadth of protection;however,poor immunogenicity greatly hinders their *** protruding(P)domain of the norovirus(NoV),which can self-assemble into a 24-mer particle called the NoV P particle,offers an ideal antigen presentation *** this study,a multiepitope nanovaccine displaying influenza epitopes(HMN-PP)was constructed based on the NoV P particle *** amounts of HMN-PP were easily expressed in Escherichia coli in soluble *** experiments showed that the adjuvanted HMN-PP nanovaccine induced epitope-specific antibodies and haemagglutinin(HA)-specific neutralizing antibodies,the antibodies could persist for at least three months after the last ***,HMN-PP induced matrix protein 2 extracellular domain(M2e)-specific antibody-dependent cell-mediated cytotoxicity,CD4^(+)and CD8^(+)T-cell responses,a nucleoprotein(NP)-specific cytotoxic T lymphocyte(CTL)*** results indicated that the combination of a multiepitope vaccine and self-assembled NoV P particles may be an ideal and effective vaccine strategy for highly variable viruses such as IV and SARS-CoV-2.

关键词： self-assemble norovirus protruding(NoV P)particle influenza virus multiepitope nanovaccine

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Degradation of amyloid β-peptides catalyzed by nattokinase in vivo and in vitro

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Food Science and Human Wellness 2023年第5期12卷 1905-1916页

作者： Aixin Ni He Li Ruya Wang Rentong Sun Yingjiu Zhang Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education Jilin UniversityChangchun 130012China School of Life Science Jilin UniversityChangchun 130012China

Amyloid-β 1-42(Aβ42)plays a pivotal role in Alzheimer disease(AD)pathogenesis. Peripheral clearance of Aβ42 largely affects its level in the brain and affects AD progression. Although nattokinase(NK)degrades Aβ40, the details of NK's capture of various Aβ species and reduction of plasma Aβ42/Aβ40 are uncharacterized. In this study, the Aβ42/Aβ40-degrading ability of NK was investigated using five Aβs and AD model mice. The C-terminal region of Aβ42/Aβ40(Gly29 to Val40)was primarily required for NK capture, and the integrated conformation in Aβ42/Aβ40 aggregates was a more efficient target for NK catalysis. Further, suspended Aβ42/Aβ40 oligomers were more easily captured by NK than suspended Aβ42/Aβ40 fibrils, while deposited Aβ42/Aβ40 fibrils recruited more NK than deposited Aβ42/Aβ40 oligomers. Although most NK was likely lost during NK uptake and/or entry into the blood, a small fraction of NK showed good plasma Aβ42/Aβ40-degrading efficacy after entering the blood due to NK's stability in the plasma of AD mice for at least 9 days. It was concluded that oral administration of NK is a feasible approach for peripheral Aβ42/Aβ40 clearance. This implies that NK might serve as an anti-Aβ42 agent for the treatment of Aβ42/Aβ40-related diseases such as AD.

关键词： Nattokinase Amyloidβ-peptide Degradation Catalyze Alzheimer disease

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Permeability and selectivity synergistically enhanced nanofluidic membrane for osmotic energy harvesting

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Carbon Energy 2024年第8期6卷 14-24页

作者： Jundong Zhong Tingting Xu Hongyan Qi Weibo Sun Shuang Zhao Zhe Zhao Yirong Sun Youliang Zhu Jianxin Mu Haibo Zhang Xuanbo Zhu Zhenhua Jiang Lei Jiang National and Local Joint Engineering Laboratory for Synthetic Technology of High‐Performance Polymer Jilin UniversityChangchunJilinChina State Key Laboratory of Supramolecular Structure and Materials Jilin UniversityChangchunJilinChina Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education Jilin UniversityChangchunJilinChina CAS Key Laboratory of Bio‐inspired Materials and Interfacial Science Technical Institute of Physics and ChemistryChinese Academy of SciencesBeijingChina

For the porous‐membrane‐based osmotic energy generator,the potential synergistic enhancement mechanism of various key parameters is still controversial,especially because optimizing the trade‐off between permeability and selectivity is still a ***,to construct a permeability and selectivity synergistically enhanced osmotic energy generator,the twodimensional porous membranes with tunable charge density are prepared by inserting sulfonated polyether sulfone into graphene *** of charge density and pore size on the ion transport are explored,and the ionic behaviors in the channel are calculated by numerical *** mechanism of ion transport in the process is studied in depth,and the fundamental principles of energy conversion are *** results demonstrate that charge density and pore size should be matched to construct the optimal ion *** collaborative enhancement strategy of permeability and selectivity has significantly improved the output power in osmotic energy generation;compared to the pure graphene oxide membrane,the composite membrane presents almost 20 times improvement.

关键词： blue energy generator charge tunable graphene oxide ion transport layered nanocomposites

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Engineered built-in electric fields in Cu^(0)/CuO_(x)nanozyme-decorated silicon nanodisks for the degradation of phenols and dyes

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Nano Research 2025年第3期18卷 142-152页

作者： Sijun Huang Tianqi Zhang Shiyin Hong Dechen Zhang Yanping Zhao Miao Li Ying Wang Xuewu Liu Yi Guo Li Xu Key Laboratory for Molecular Enzymology and Engineering Ministry of EducationSchool of Life SciencesJilin UniversityChangchun 130012China Department of Nanomedicine Houston Methodist Academic Institute6670 Bertner AvenueHoustonTX 77030USA State Key Laboratory of Rare Earth Resource Utilization Changchun Institute of Applied ChemistryChinese Academy of SciencesChangchun 130022China

Laccase has demonstrated potential for the treatment of hazardous pollutants;however,its widespread application is hindered by stability *** contrast,nanozymes,with their remarkable stability,present a promising *** this study,we developed silicon-anchored Cu^(0)/CuO_(x)nanozymes exhibiting laccase-like activity for the oxidation removal of phenols and *** valence states of the copper species,which emerged through spontaneous oxidation,played a crucial role in creating a heterogeneous interface,exerting a significant impact on the catalytic efficacy of the copper *** employing density functional theory(DFT)calculations,we revealed that the existence of a local built-in electric field(BIEF)among the heterogeneous components facilitated the cyclic consumption of Cu^(0)and the migration of lattice *** dynamic interplay modulated the levels of Cu^(+)and oxygen vacancies(OVs),thereby allowing for sustained catalytic performance within a defined *** findings underscore the importance of valence engineering in the rational design of nanozymes and highlight their potential as efficient catalysts for advancing environmental sustainability.

关键词： nanozyme laccase mimic built-in electric field oxidation removal

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Design and fabrication of highly hydrophilic magnetic material by anchoring L-cysteine onto chitosan for efficient enrichment of glycopeptides

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Chinese Chemical Letters 2023年第3期34卷 148-152页

作者： Binfen Zhao Wenhui Xu Jiutong Ma Qiong Jia College of Chemistry Jilin UniversityChangchun 130012China Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education School of Life SciencesJilin UniversityChangchun 130012China

Hydrophilic interaction liquid chromatography(HILIC) has been recognized as an effective strategy for glycopeptide enrichment. Hydrophilic materials pave the way to solve the limit of low enrichment capacity and poor selectivity. The present study is the first attempt to combine chitosan(CS) and L-cysteine(L-Cys) to design a novel hydrophilic material focusing on glycopeptide enrichment. CS containing a large number of hydrophilic amino and hydroxyl groups has unique chemical properties, which makes it a very attractive biomaterial for glycopeptide enrichment. The excellent hydrophilicity of zwitterionic molecule L-Cys inspires the idea of anchoring L-Cys onto CS to design a novel hydrophilic material(named as Fe_(3)O_(4)@CS@Au-L-Cys) for the capture of low abundance glycopeptides. To be specific, Au nanoparticles(Au NPs) was introduced into CS-coated Fe_(3)O_(4)via electrostatic interaction and served as bridges to anchor L-Cys onto the surface of CS through strong Au-S bond interaction. The prepared Fe_(3)O_(4)@CS@AuL-Cys exhibited strong affinity, low detection limit(0.5 fmol/μL HRP), high selectivity(HRP/BSA with a molar ratio of 1:1000) for glycopeptides. Moreover, successful application of glycopeptide enrichment in human serum and saliva by Fe_(3)O_(4)@CS@Au-L-Cys was achieved. A satisfactory data set indicates that Fe_(3)O_(4)@CS@Au-L-Cys has promising potential in the application of glycopeptide enrichment in real complex bio-samples and for related glycoproteome research.

关键词： Chitosan L-cysteine Hydrophilic material Glycopeptide Enrichment

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Hydrophilic adamantane derivatives engineered β-cyclodextrin-based self-assembly materials for highly efficient enrichment of glycopeptides

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Chinese Chemical Letters 2023年第3期34卷 283-287页

作者： Ping Li Zheng Li Dawei Zhang Qiong Jia College of Chemistry Jilin UniversityChangchun 130012China Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education College of Life SciencesJilin UniversityChangchun 130012China

β-Cyclodextrin(β-CD) based materials have attracted great attention in the separation of hydrophilic glycopeptides due to the abundant hydroxyl groups in its exterior. However, the current materials based on β-CD generally has complex synthesis process and harsh experimental conditions, on the other hand,the interior cavity of β-CD is hydrophobic and is harmful to capture glycopeptides. Herein, a novel hydrophilic material based on β-CD was engineered via a self-assembly process utilizing L-cysteine(L-Cys)or glutathione(GSH) derived adamantane for highly efficient glycopeptide enrichment. It is the first attempt to make use of the hydrophobic interior cavity of β-CD for hydrophilic glycopeptide capture. Taking advantages of strong hydrophilicity and superparamagnetism, the as-prepared materials possess low detection limit, high selectively, and excellent reusability when employed to glycopeptide enrichment. In addition, the feasibility of the hydrophilic material based on β-CD was verified by enriching glycopeptides from human serum and saliva samples. This study provides a heuristic strategy for the application of β-CD-based self-assembly materials in the enrichment of glycopeptides. Importantly, this strategy certified a possible that the change of glycopeptide enrichment sites through host-guest interaction between β-CD and adamantane derivatives with different functional groups.

关键词： β-Cyclodextrin Hydrophilic Self-assembly Enrichment Glycopeptides

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In vivo Toxicity Evaluation of a Nano-drug Delivery System Using a Caenorhabditis elegans Model System

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Chemical Research in Chinese Universities 2022年第4期38卷 1018-1024页

作者： HAN Wenzhao LI Hui YU Xiaoxuan KE Junfeng GUO Fng WANG Liping Key Laboratory for Molecular Enzymology and Engineering Ministry of EducationSchool of Life SciencesEngineering Laboratory for AIDS VaccineJilin UniverstiyChangchun130012P.R.China

Drug carrier materials need to possess good biological ***,most biosafety evaluation studies use rodent animal models,including rats and ***,the cost of raising these animals is relatively high and the experimental period is *** elegans(***)presents an ideal toxicological evaluation model due to its simple structure,easy cultivation,short life cycle,and evolutionary *** this paper,we used *** to test the biological safety of our pH-responsive carrier system(FFPFF self-assembling into a nanosphere structure,FFPFF Nps),which was designed for anti-tumor drug *** results showed that exposure to high doses of FFPFF Nps did not have a significant impact on the survival rate,growth,development,movement,and reproduction of *** preliminary evaluation of the overall biological model of *** shows that FFPFF Nps has good biological safety and warrants further study.

关键词： Toxicity Caenorhabditis elegans Nano drug delivery system

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Confining carbon dots in amino-functionalized mesoporous silica:n→π^(*) interaction triggered deep-red solid-state fluorescence

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Nano Research 2023年第3期16卷 4170-4177页

作者： Hongyue Zhang Qingyi Li Shuo Wang Xiaowei Yu Bolun Wang Guangrui Chen Li Ren Jiyang Li Mingxing Jin Jihong Yu State Key Laboratory of Inorganic Synthesis and Preparative Chemistry College of ChemistryJilin UniversityChangchun 130012China Institute of Atomic and Molecular Physics Jilin UniversityChangchun 130012China Key Laboratory for Molecular Enzymology and Engineering The Ministry of EducationJilin UniversityChangchun 130012China International Center of Future Science Jilin UniversityChangchun 130012China

Deep-red and near-infrared emissive carbon dots(CDs)are highly desired for bioimaging,especially in deep tissue imaging,but they are extremely rare and the known ones usually suffer from low-efficient fluorescence in water and aggregation-induced fluorescence quenching in solid *** this work,CDs with intriguing solvent-dependent and two-photon fluorescence emissions have been prepared by a facile solvothermal *** characterizations reveal that there is an n→π*interaction between the carboxyl functional groups on CDs and the electron donor groups in solvent,which leads to the increase of energy density of CDs and the decrease of energy level,resulting in the red shift of luminescence with enhanced electron donating ability of *** by this finding,mesoporous silica nanoparticles(MSNs)with suitable pore size and low biological toxicity are modified by amino groups to confine CDs,thus the deep-red fluorescence emission is achieved both in solid state and in water facilitated by the n→π*interaction of *** as-prepared CDs@EDA-MSN composite exhibits high-efficient fluorescence with 650 nm wavelength,low toxicity,and good biocompatibility,which endow them a promising application in bio-imaging.

关键词： carbon dots mesoporous silica deep-red fluorescence donor-acceptor modulation bioimaging

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