INTERNATIONAL JOURNAL OF FOOD PROPERTIES
·
2018
·
IF 1.845
Zhibing Wu,Fengwen Huang,Yutao Chen,Hong Xu,Manjunath D. Meti,Yu Fan,Qingguo G. Han,Haifeng Tang,Zhendan He,Zhangli Hu
The interaction of trypsin with acteoside was studied using ultraviolet visible absorption, fluorescence, synchronous fluorescence, circular dichroism techniques, along with molecular docking method. The fluorescence experiments indicated that acteoside quenched the intrinsic fluorescence of trypsin via a combined quenching process (static and dynamic quenching). The binding constant of acteoside to trypsin obtained was 2.50 × 105 L mol−1 at 298 K and the number of binding site was about one under the same experimental condition. The thermodynamic functions ΔH° and ΔS° of the binding process were 8.79 kJ mol−1 and 132.58 J mol−1 K−1, respectively, which indicated that the hydrophobic force was the main acting force between them. Ultraviolet–visible, synchronous fluorescence together with circular dichroism spectra studies demonstrated that the interaction of acteoside with trypsin lead to a loosening and unfolding of the protein backbone with partial β-sheet structures being transformed into α-helix structures. All these experimental results were validated and explained reasonably by docking studies. And the molecular docking results further illustrated that besides hydrophobic forces, hydrogen bonds also played an important role in the stabilization of the acteoside–trypsin complex. Results from this study should be helpful to make full use of acteoside in the food industry and be useful to the design of the drugs for the diseases related to trypsin.
研究领域:
生物化学
·
药物设计
·
食品科学
✓ 已验证
产品关联
DOI
高影响因子
ANALYST
·
2025
·
IF 3.3
Jiahui Lai,Danni Wang,Xiuqin Sheng,Chuan-Fan Ding,Yinghua Yan
In large-scale glycoproteomics studies, reducing the experimental duration and enhancing digestion efficiency are crucial. In this work, a magnetic nanomaterial featuring a surface rich in hydrophilic groups and uniformly distributed trypsin (Fe3O4@DOPA/PEI@Au@DNA TET-trypsin) was developed for efficient digestion of proteins and specific enrichment of glycopeptides. The material surface exhibits a regular distribution of trypsin coupled with abundant hydrophilic groups. Thus, the material can demonstrate excellent performance in experimental studies related to glycoproteomics. Fe3O4@DOPA/PEI@Au@DNA TET-trypsin showed an excellent limit of detection (0.001 amol·μL−1) and high selectivity for glycopeptides (BSA : HRP = 5000 : 1) in a short reaction time, and it showed good performance in the processing of practical samples. Gene Ontology (GO) analysis suggests that complement activation, extracellular space, antioxidant activity and complement binding may be associated with tuberculosis. These analytical results provide new insights for determining and clarifying the pathogenesis and pathology related to tuberculosis.
研究领域:
蛋白质组学
·
生物化学
·
医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TRANSACTIONS OF TIANJIN UNIVERSITY
·
2015
·Vol. 21
·期 5
·433-439
·
IF 0
Zhu Hongji,Wang Shipeng,Tian Li,Zhang Hua
Antioxidant peptides of flatfish skin protein hydrolyzed by four enzymes(Papain, Pepsin, Trypsin and Neutrase, respectively)were investigated. The Trypsin hydrolysate obtained by hydrolysis exhibited the highest 1,1-dipheny-l-2-picrylhydrazyl(DPPH)radical scavenging activity(DRSA)compared with other hydrolysates. Response surface method ology(RSM), based on Box-Behnken design, was used to study the influence of hydrolysis conditions on the DRSA. The optimal hydrolysis conditions were as follows: pH 7.38, temperature 48.2°C and enzyme/substrate(E/S)ratio 2 840 U/g. Under these conditions, the maximum DRSA was(22.85 ± 0.57)%,. The experimental values agreed with the value (23.09%,)predicted by the model within a 95%, confidence interval. By using gel filtration chromatography and reversed-phase high performance liquid chromatography(RP-HPLC), antioxidant peptide(D2-P)was isolated from flatfish skin protein hydrolysates(FSPH)and could exhibit a(54.28 ± 1.37)%, scavenging activity on DPPH radical at the concentration of 5 mg/mL. This is the first report of a scientific basis for the preparation of antioxidant peptides from flatfish skin. The results suggested that the antioxidant peptides can be exploited into functional foods or used as a novel source of nutraceuticals.
研究领域:
食品科学
·
生物化学
✓ 已验证
产品关联
DOI
ANALYTICAL BIOCHEMISTRY
·
2013
·
IF 2.582
Wenao Min,Shumin Cui,Weiping Wang,Jianrong Chen,Zhide Hu
An efficient and durable online capillary immobilized trypsin microreactor was successfully established to study the enzyme kinetics of trypsin and screen its inhibitors from natural extracts through capillary electrophoresis (CE). In this procedure, trypsin was immobilized on the inner wall at the inlet of the capillary treated with 3-aminopropyltrimethoxy silane (3-APTES), producing a trypsin microreactor via cross-linking of glutaraldehyde with 3-APTES and trypsin. The rest of the capillary was selected as a channel for separating the generated product and unreacted substrate of the trypsin enzymatic reaction . The parameters affecting the separation efficiency and activity of immobilized trypsin were evaluated systematically. The optimized conditions were as follows: 50 mM Tris–HCl (pH 8.0), 15 kV, 37 °C, 10 mM substrate, incubation for 2 min. Under optimal conditions, separation of the product and substrate was achieved through CE within 3.5 min. The obtained results of Michaelis constant , inhibition kinetics constant, and half-maximal inhibitory concentration for the immobilized trypsin using benzamidine hydrochloride hydrate as a model inhibitor were 1.56, 1.79 and 3.98 mM, respectively. The proposed method was successfully applied for screening of trypsin inhibitors from 19 kinds of natural extracts.
研究领域:
生物化学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2022
·
IF 8.008
Yuanyuan Cai,Tao Dong,Xin Zhang,Aihua Liu
To regulate nanostructure synthesis is of crucial importance for developing various applications, including catalysis, bioanalysis, and optical devices. Herein, the morphology and peroxidase (POD)-mimicking activity of peptide-templated copper nanoassemblies (Cu NAs) are regulable with peptide types. The Cu NAs templated with peptide containing single cysteine are uniform nanoclusters with strong POD-like activity. However, the Cu NAs templated with peptide containing two cysteines are fusiform-like with very weak POD-like activity. Unexpectedly, the POD-like activity of Cu NAs templated with peptide containing two cysteines with lysine between the cysteines is significantly enhanced when trypsin is incubated, which is unchanged for the Cu NAs templated with peptide containing two cysteines without lysine between the cysteines. The remarkably enhanced POD-mimicking activity originates from trypsin specifically shearing the peptide bond on the lysine, thereby allowing the aggregated Cu NAs to unravel into individual nanoclusters. Therefore, a robust colorimetric sensing platform was constructed for sensitive and selective detection of trypsin, which showed a linear concentration range of 3–1000 nM and a detection limit of 0.82 nM (S/N = 3). More interestingly, featured by trypsin inhibitor restraining trypsin activity, it enabled us to screen trypsin inhibitors as well. Subsequently, the developed assay was applied to detect trypsin in serum samples with good accuracy and reproducibility. Thus, this strategy shows great potential application in the clinic for diagnosis of trypsin-indicating diseases as well as the screening of trypsin inhibitor-based anti-cancer drugs.
研究领域:
纳米技术
·
生物化学
·
分析化学
·
生物医学工程
·
药物发现
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH
·
2020
·Vol. 27
·期 12
·13409-13416
·
IF 3.056
Wang Xiaofang,Hui Hongjie,Yu An,Jiang Ziyang,Yu Hongyan,Zou Luyi,Teng Yue
Triclosan (TCS), a broad-spectrum antibacterial agent, exhibits a high exposure in the environment. However, the residual TCS in the environment poses a potential risk to human health. In this study, spectroscopic methods, molecular docking and animal experiment were conducted to completely understand the interaction between trypsin and TCS. The formation of the TCS-trypsin complex was spontaneously achieved through hydrogen bonds and Van der Waals forces with a binding constant ( K a ) between 10 3 and 10 4 L mol −1 . In addition, the trypsin activity in fish intestine was inhibited by TCS exposure, revealing the potentially negative effects of TCS on metabolism. The results might be explained by changes in the conformation of the trypsin, inducing the content of unordered coil increasing significantly (from 36.2% to over 80%). This work provides useful information for assessing the toxicity of TCS at the molecular level.
研究领域:
环境科学
·
生物化学
·
毒理学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF ANALYTICAL CHEMISTRY
·
2026
·
IF 1.5
Zhu Yong,Jiang Cuifeng,Wei Zijie,Han Wanting,Huo Min
Detection of trypsin is crucial for human health. In this work, bovine serum albumin-protected osmium-silver nanoclusters were prepared. The silver content in the nanoclusters could be regulated by controlling the reagent concentration. Ultraviolet-visible absorption spectra of 3,3',5,5'-tetramethylbenzidine and its oxidized form were used to investigate the catalytic activity of the nanoclusters. All nanoclusters displayed peroxidase-like activity. Especially, the bovine serum albumin-protected osmium-silver nanoclusters with 20% silver content showed the highest catalytic activity. In addition, trypsin effectively increased the catalytic activity of bovine serum albumin-protected osmium-silver nanoclusters due to the interaction between trypsin and bovine serum albumin. Based on this phenomenon, a novel method for selective detection of trypsin was developed with a limit of detection of 5 ng/mL. This method shows promise for the detection of trypsin in fetal bovine serum with an acceptable recovery rate.
✓ 已验证
DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2011
·
IF 4.311
Chunhe Yao,Li Qi,Wenbin Hu,Fuyi Wang,Gengliang Yang
A new kind of immobilized trypsin reactor based on sub-micron skeletal polymer monolith has been developed. Covalent immobilization of trypsin on this support was performed using the epoxide functional groups in either a one- or a multi-step reaction. The proteolytic activity of the immobilized trypsin was measured by monitoring the formation of N -α-benzoyl- l -arginine (BA) which is the digestion product of a substrate N -α-benzoyl- l -arginine ethyl ester (BAEE). Results showed that the digestion speed was about 300 times faster than that performed in free solution. The performance of such an enzyme reactor was further demonstrated by digesting protein myoglobin . It has been found that the protein digestion could be achieved in 88 s at 30 °C, which is comparable to 24 h digestion in solution at 37 °C. Furthermore, the immobilized trypsin exhibits increased stability even after continuous use compared to that in free solution. The present monolithic enzyme-reactor provides a promising platform for the proteomic research.
研究领域:
生物化学
·
蛋白质组学
·
酶工程
·
分析化学
✓ 已验证
DOI
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2021
·Vol. 413
·期 20
·4939-4945
·
IF 4.157
Guan Shulin,Yue Jing,Sun Weihan,Xu Weiqing,Liang Chongyang,Xu Shuping
A highly sensitive trypsin sensing system in serum was developed by using an anodic alumina oxide (AAO)-based, trypsin substrate-decorated hybrid ion permeation membrane. Owing to the trypsin-triggered peptide hydrolyzation reaction, the surface electrical feature of the peptide-decorated hybrid ion membrane changed. The electric double layer effect reduces the effective ion current diameter in the AAO nano unit, so that the ion current rectification ratio will be enhanced, realizing the quantitative detection of trypsin. The lowest detection concentration can be achieved as low as 0.1 pM. This method is no need for sample pre-preparation, easy to operate, highly sensitive, and also applicable to other enzyme evaluation systems by changing corresponding substrates. This study provides a new idea for selective measurements of proteases in complex biological samples. Graphical abstract
研究领域:
生物化学
·
医学检测
·
纳米技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Hazardous Materials Advances
·
2021
·
IF 0
Luo Li,Yi Li,Chunhu Tan,Tingting Zhang,Xiaoli Xin,Weihua Li,Jiansheng Li,Rui Lu
The mechanism of GO/rGO interaction with trypsin is investigated using multiple methods. UV-Vis reveals that trypsin has a maximum absorption at 278 nm, the absorption intensity increased demonstrating GO/rGO alters the solvent environment of trypsin and exposes the aromatic chromogenic groups. ITC experiments with GO and trypsin provide a negative ΔH of -25.98 KJ mol −1 and the association constant Ka of 4.829 × 10 −4 . The negative ΔS of -23.64 J mol −1 ∙K −1 is further given by the Gibbs–Helmholtz equation. Combining with the ζ potential result, the driving forces for binding include hydrogen bonding, electrostatic interaction forces and van der Waals forces. Fluorescence spectroscopy shows that GO/rGO can significantly quench the fluorescence intensities of trypsin. The equilibrium constant Kd of GO/ rGO-24h / rGO-48h is 4.41±0.07, 5.82±0.53, 7.24±0.25, respectively, which means the reduced content of oxygen-containing groups can weaken the binding forces between GO/rGO and trypsin. CD show that even low concentrations of GO/rGO affect the secondary structure of trypsin, mainly decreasing the α -helix and β -fold content while increasing the β -turn and random coil content of trypsin. Overall, GO/rGO can effectively interact with trypsin, the oxidation degree of GO/rGO can affects the combination of trypsin and further influence on protein conformation and function.
研究领域:
生物化学
·
分子生物学
·
蛋白质科学
✓ 已验证
DOI
ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Junpeng Lv,Jingjing Xu,Yiting Chen,Wenqi Tang,Wenhui Xu,Mengge Wang,Xiyao Zhang,Ziluan Zhu,Songmin Chen,Qiong Hu,Li Niu
Although polysaccharides are ubiquitous in nature, biodegradable, and cheap, there is no report illustrating their use as functional materials for signal amplification in the biosensing of enzyme activities. Herein, we report the development of a polysaccharide-amplified electrochemical biosensor for the highly sensitive detection of trypsin activity. The substrate peptides without carboxyl sites are attached to the electrode surface via the N-terminus. Such that, the tryptic cleavage of the peptide layer gives rise to a free carboxyl site at the C-terminus of each truncated fragment, and the carboxyl-carrying polysaccharide chains are then recruited via the carboxylate-Zr(IV)-carboxylate linkage, followed by the decoration of redox labels via the cross-linking between the cis-diol sites of polysaccharide chains and the phenylboronic acid group. Through boronate cross-linking, each polysaccharide chain can be decorated with hundreds of redox labels, leading to the significantly amplified electrochemical detection of trypsin activity. The polysaccharide-based signal amplification is advantageous due to its simple operation, low cost, and mild reaction conditions. With alginic acid as the proof-of-concept polysaccharide, a detection limit of 103.5 μU/mL (∼0.41 ng/mL) has been obtained for trypsin detection. The cleavage-based recognition enabled the selective discrimination of trypsin against other proteases and proteins. In addition, the potential use of electrochemical biosensors in inhibitor screening, together with the interrogation of trypsin activity in urine samples, has been validated. Collectively, the results clearly show that the polysaccharide-amplified electrochemical biosensor is highly applicable to the highly sensitive detection of trypsin activity.
研究领域:
生物传感器
·
电化学
·
酶活性检测
·
分析化学
·
纳米生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL BIOCHEMISTRY
·
2018
·
IF 2.275
Lide Gu,Wanli Yan,Shu Liu,Wei Ren,Mingsheng Lyu,Shujun Wang
A novel screening method for protein aptamer selection was developed in this study. Aptamers with high affinity and specificity to the surface recombinant antigen of Helicobacter pylori (HP-Ag) and to tumor markers carcinoembryonic antigen (CEA), cancer antigen 125 (CA125) and cancer antigen 19-9(CA19-9) were screened using trypsin enhanced screening method. Briefly, the target proteins above were immobilized onto 96-well polystyrene plates and incubated with a single-stranded DNA (ssDNA) library for aptamer selection. Then, trypsin was introduced to digest the proteins and obtain ssDNA that bound to the target proteins with high specificity. The concentration of ssDNA that shed from protein-ssDNA complexes was detected. After sequencing, the enrichment of target-specific aptamers was monitored and the affinity of each aptamer was analyzed. Urea, which has been reported in other article, was used to compare with trypsin. The results revealed that trypsin was more effective than urea for protein aptamer selection. The protocol used in this study provided a novel method for generating aptamers.
研究领域:
分子生物学
·
生物技术
·
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）