ANALYTICA CHIMICA ACTA
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2016
·
IF 4.712
Kaijia Xu,Yuzhi Wang,Yixue Li,Yunxuan Lin,Haibao Zhang,Yigang Zhou
Novel poly(deep eutectic solvent) grafted silica-coated magnetic microspheres (Fe 3 O 4 @SiO 2 -MPS@PDES) were prepared by polymerization of choline chloride-itaconic acid (ChCl-IA) and γ-MPS-modified magnetic silica composites, and were characterized by vibrating sample magnetometer (VSM), Fourier transform infrared spectrometry (FT-IR), X-ray photoelectron spectra (XPS), thermal gravimetric analysis (TGA) and transmission electron microscope (TEM). Then the synthetic Fe 3 O 4 @SiO 2 -MPS@PDES microspheres were applied for the magnetic solid-phase extraction (MSPE) of trypsin for the first time. After extraction, the concentration of trypsin in the supernatant was determined by a UV–vis spectrophotometer. Single factor experiments were carried out to investigate the effects of the extraction process, including the concentration of trypsin, the ionic strength , the pH value, the extraction time and the temperature. Experimental results showed the extraction capacity could reach up to 287.5 mg/g under optimized conditions. In comparison with Fe 3 O 4 @SiO 2 -MPS, Fe 3 O 4 @SiO 2 -MPS@PDES displayed higher extraction capacity and selectivity for trypsin. According to the regeneration studies, Fe 3 O 4 @SiO 2 -MPS@PDES microspheres can be recycled six times without significant loss of its extraction capacity, and retained a high extraction capacity of 233 mg/g after eight cycles. Besides, the activity studies also demonstrated that the activity of the extracted trypsin was well retained. Furthermore, the analysis of real sample revealed that the prepared magnetic microspheres can be used to purify trypsin in crude bovine pancreas extract . These results highlight the potential of the proposed Fe 3 O 4 @SiO 2 -MPS@PDES-MSPE method in separation of biomolecules.
研究领域:
生物化学
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分析化学
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DOI
PubMed
高影响因子
ACS Sustainable Chemistry & Engineering
·
2013
·
IF 0
Longhua Zou,Wei Qi,Renliang Huang,Rongxin Su,Mengfan Wang,Zhimin He
We report a green synthesis of novel gold nanoparticle–nanocluster composite nanostructures directly using trypsin as linking and reducing agents. Size exclusion chromatography (SEC) and transmission electron microscopy (TEM) reveals that the as-prepared gold nanocomposite (gold nanoparticles-trypsins-nanoclusters, GNPs-Trys-GNCs) is composed of GNPs (gold nanoparticles) with an average diameter of 5.5 nm and the GNCs (gold nanoclusters, about 1 nm)-embedded trypsins (Trys-GNCs), which are attached to the surface of the GNPs. The specific amino acids in the trypsin molecule, like cysteine, methionine, and tyrosine, combined with the unique spatial structures, enable trypsins to bind and reduce the AuCl4– ions, simultaneously forming GNPs and GNCs in one-pot synthesis. Similar to pure GNPs, the GNPs-Trys-GNCs nanocomposite also exhibits an intense surface plasmon resonance (SPR) absorbance at 520 nm. However, it shows an obvious different optical property in the interaction with Pb2+ ions. In the presence of Pb2+ ions, an increased intensity and a slight red-shift of the SPR peak for the GNPs-Trys-GNCs nanocomposite in the UV–vis spectra were observed, while a decreased intensity and large red-shift for pure GNPs were observed in previous studies. Moreover, we found that the SPR intensity linearly increased with Pb2+ concentration from 1.6 to 32.3 μM (R2 = 0.9731). In addition, high-level Pb2+ ions would induce the aggregation of GNPs-Trys-GNCs nanocomposite accompanied by the formation of precipitate. The unique structure and optical property of the GNPs-Trys-GNCs nanocomposite enable it to be used in heavy metal ions sensing and elimination.
研究领域:
纳米生物学
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环境科学
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分析化学
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DOI
Analytical Methods
·
2023
·Vol. 15
·期 29
·3586-3591
·
IF 3.1
Li Zhang,Jiyan Huang,Mixue Chen,Hongmei Huang,Yi Xiao,Ronghua Yang,Youyu Zhang,Xiaoxiao He,Kemin Wang
Amphiphilic aggregation-induced emission (AIE) molecules show superior potential for fabricating novel ultrasmall nanoprobes. Here, an anionic dipyridyl tetraphenylethene (TPE) derivative is rationally designed and a super-small self-assembled AIEgen nanoprobe (TPE-2Py-SO3NaNPs, ca. 2.48 nm) is thus conveniently constructed for the supersensitive detection of protamine and trypsin. In HEPES/DMSO solution (8 : 2, v/v, pH = 7.4), negatively charged TPE-2Py-SO3NaNPs exhibited an AIE effect in the presence of positively charged protamine, presenting a fluorescence enhancement at 498 nm together with a large Stokes shift of 150 nm and a low detection limit of 8.0 ng mL−1. In addition, the in situ formed TPE-2Py-SO3Na/protamine nanocomposite can be dissociated by trypsin due to the highly selective degradation of protamine via enzymatic hydrolysis, achieving a detection limit for trypsin as low as 5.0 ng mL−1.
研究领域:
生物化学
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分析化学
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医学检测
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2014
·
IF 4.517
Gong Cheng,Ping Chen,Zhi-Gang Wang,Xiao-Jing Sui,Ji-Lin Zhang,Jia-Zuan Ni
A simple, fast, efficient, and reusable microwave-assisted tryptic digestion system which was constructed by immobilization of trypsin onto porous core-shell Fe 3 O 4 @fTiO 2 microspheres has been developed. The nanostructure with magnetic core and titania shell has multiple pore sizes (2.4 and 15.0 nm), high pore volume (0.25 cm 3 g −1 ), and large surface area (50.45 m 2 g −1 ). For the proteins, the system can realize fast and efficient microwave-assisted tryptic digestion. Various standard proteins (e.g., cytochrome c (cyt-c), myoglobin (MYO), β-lactoglobulin (β-LG), and bovine serum albumin (BSA)) used can be digested in 45 s under microwave radiation, and they can be confidently identified by mass spectrometry (MS) analysis; even the concentration of substrate is as low as 5 ng μL −1 . Furthermore, the system for the 45 s microwave-assisted tryptic digestion is still effective after the trypsin-immobilized microspheres have been reused for 5 times. Importantly, 1715 unique proteins from 10 μg mouse brain proteins can be identified with high confidence after treatment of 45 s microwave-assisted tryptic digestion.
研究领域:
生物化学
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分析化学
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蛋白质组学
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DOI
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2026
·
IF 4.8
Xinxin Liu,Tianjiao Li,Jihong Wang,Xian Wang,Liqian Niu,Xinyu Fan,Xin Guan,Shiyue Niu,Xin Liu,Shuyun Bi
Lysozyme and trypsin were separately used as fluorescent probes for the sensitive detection of doxycycline, owing to the stable intrinsic fluorescence, excellent biocompatibility, low toxicity, and low cost of the two proteins. Fluorescence quenching of both lysozyme and trypsin induced by doxycycline was verified to be static by the stern-Volmer equation and time-resolved fluorescence measurements. Synchronous fluorescence and three-dimensional (3D) fluorescence spectra of the proteins exhibited no obvious peak shift upon the addition of doxycycline, indicating that the microenvironment surrounding tyrosine (Tyr) and tryptophan (Trp) residues was barely disturbed. Circular dichroism (CD) and Fourier transform infrared (FT-IR) spectra demonstrated that doxycycline modified the secondary structure of the proteins. Binding constants ( K a ) and binding sites ( n ) were obtained at various temperatures. Molecular docking and thermodynamic analysis revealed that electrostatic forces played a dominant role in the intermolecular interactions. The static quenching process was accompanied by non-radiative energy transfer. Interference experiments demonstrated that the proposed method exhibited good selectivity. For the lysozyme and trypsin systems, the linear calibration equations were F 0 / F = 0.1005 c + 0.9911 and F 0 / F = 0.0884 c + 0.9708 over the concentration ranges of 6.2–1.6 × 10 4 ng mL −1 and 8.2–1.8 × 10 4 ng mL −1 , with limits of detection (LOD) of 0.58 and 0.89 ng mL −1 , respectively. The proposed method was applied to the determination of doxycycline in fish, pork, and chicken samples, and satisfactory recoveries verified its accuracy and feasibility
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DOI
PubMed
高影响因子
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
·
2026
·
IF 2.5
Zhongguang Lu,Xuan Wang,Yanran He,Hua Zeng,Chengwei Zhou,Yexun Liu,Ziyi Huang,Qimeng Fan
Betulinic acid (BA) is a naturally occurring pentacyclic triterpenoid with diverse pharmacological activities. This study systematically investigated the binding mechanisms of BA with pepsin (PEP), trypsin (TRY), and α-chymotrypsin (CHY). Steady-state and synchronous fluorescence, circular dichroism (CD), Fourier transform infrared (FTIR) spectroscopy, molecular docking, and molecular dynamics (MD) simulations were employed to characterize the interactions, fluorescence quenching, and conformational changes. BA quenched the intrinsic fluorescence of all three enzymes in a concentration- and temperature-dependent manner; PEP underwent dynamic quenching, whereas TRY and CHY exhibited static or mixed-mode quenching. Binding analyses revealed a single dominant site on each enzyme, with the highest affinity observed for CHY. Thermodynamic analysis indicated that hydrophobic interactions predominantly drove BA–PEP binding, whereas electrostatic interactions and hydrogen bonding were critical for BA–TRY and BA–CHY complexation. FTIR confirmed that BA bound to all three enzymes via synergistic hydrogen bonding and hydrophobic interactions, inducing secondary structural alterations. Molecular docking revealed that BA stably occupied the active sites of each enzyme, forming enzyme-specific binding modes. MD simulations (RMSD, Rg, SASA, hydrogen bond analysis, and MMPBSA) validated the dynamic stability of the complexes, with binding free energies of −13.90, −20.14, and −23.08 kcal/mol for BA–PEP, BA–TRY, and BA–CHY, respectively, consistent with the experimental affinity trend. These findings elucidate the molecular mechanisms by which BA modulates digestive proteases, providing insights into its gastrointestinal behavior and bioactivity.
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DOI
PubMed
高影响因子
PROCESS BIOCHEMISTRY
·
2024
·
IF 3.7
Xuyang Chen,Hao Liu,Chao Lei,Yirui Lin,Bi Shi,Yunhang Zeng
Bating animal hides with trypsin is an essential step for making leathers. But the mechanism why the calcium in the hide influences the bating performance remains unclear. Here, the enzyme activity of trypsin was determined using N-alpha-benzoyl-L-arginine ethyl ester, bovine albumin (BSA) and collagen fibres as substrates in the presence and absence of calcium salts. The interactions of calcium with trypsin, BSA and collagen were analysed by fluorescence spectroscopy and molecular docking. Data reveal that calcium which binds to BSA and collagen improves the structural stability and enzyme resistance of hide proteins. As a result, a high calcium content of delimed hide is useful to obtain an intact surface of bated hide. However, a low calcium content of hide for pickling is necessary for producing leathers with high physical properties and uniform colour. According to these results, a novel bating system involving deliming, bating and redeliming operations was developed. This system provides a high calcium content of hide for bating and a low calcium content of hide for subsequent pickling and tanning, which effectively reduces the risk of hide damage and improves the physical properties and colour uniformity of leather.
研究领域:
化学
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生物化学
·
材料科学
·
皮革技术
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产品关联
DOI
高影响因子
COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS
·
2024
·
IF 4.9
Chenzhao Wang,Xiujuan Wang,Yuhao Jiang,Zhiyu Wu,Jiawei Yang,Shanshan Wei,Zhibing Wang,Guoying Sun
Trypsin is a kind of digestive enzyme capable of catalyzing the hydrolysis of proteins. Abnormal changes in the levels of trypsin can cause a series of pancreatic-related diseases. Therefore, based on the special interface interaction between green-emitting ionic liquid-based carbon dots (G-CDs) and BSA, a highly sensitive fluorescence "on-off" mechanism was developed for detecting trypsin in urine. The G-CDs were synthesized by a one-step hydrothermal approach using 2,5-dihydroxybenzoic acid (2,5-DHBA) and 1-octyl-3-methylimidazolium tetrafluoroborate ([OMIM]PF 4 ). The interaction between G-CDs and bovine serum albumin (BSA) led to the formation of the G-CDs-BSA fluorescence sensing system, which induced aggregation and fluorescence enhancement of G-CDs. The G-CDs-BSA system was disrupted under optimal incubation conditions after the addition of trypsin, and the fluorescence was quenched. According to the fluorescence quenching value (∆F) of the system, the quantitative analysis of trypsin in urine was realized and demonstrated good recoveries. This study not only developed a novel fluorescence sensing system with excellent performance for trypsin detection, but also provided a theoretical basis for the binding of carbon dot-protein systems.
研究领域:
生物化学
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医学诊断
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DOI
高影响因子
JOURNAL OF SEPARATION SCIENCE
·
2020
·Vol. 43
·期 15
·3136-3145
·
IF 2.878
Hao Zhang,Min Lu,Hui Jiang,Xu Wang,Feng-Qing Yang
In this study, a capillary electrophoresis-based online immobilized enzyme microreactor was developed for evaluating the inhibitory activity of green tea catechins and tea polyphenol extracts on trypsin. The immobilized trypsin activity and other kinetic parameters were evaluated by measuring the peak area of the hydrolyzate of chromogenic substrate S-2765. The results indicated that the activity of the immobilized trypsin remained approximately 90.0% of the initial immobilized enzyme activity after 30 runs. The value of Michaelis–Menten constant ( K m ) was (0.47 ± 0.08) mM, and the half-maximal inhibitory concentration (IC 50 ) and inhibition constant ( K i ) of benzamidine were measured as 3.34 and 3.00 mM, respectively. Then, the inhibitory activity of four main catechins (epicatechin, epigallocatechin, epicatechin gallate, and epigallocatechin gallate) and three tea polyphenol extracts (green tea, white tea, and black tea) on trypsin were investigated. The results showed that four catechins and three tea polyphenol extracts had potential trypsin inhibitory activity. In addition, molecular docking results illustrated that epigallocatechin gallate, epicatechin gallate, epicatechin, and epigallocatechin were all located not only in the catalytic cavity, but also in the substrate-binding pocket of trypsin. These results indicated that the developed method is an effective tool for evaluating inhibitory activity of catechins on trypsin.
研究领域:
分析化学
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生物化学
·
药理学
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食品科学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF BIOMOLECULAR STRUCTURE & DYNAMICS
·
2019
·
IF 3.31
Jialiang Lin,Yang Xu,Yuhan Wang,Songyang Huang,Junwei Li,Manjunath D. Meti,Xu Xu,Zhangli Hu,Johnson Liu,Zhendan He,Hong Xu
The interaction of trypsin with Gensenoside-Rg1 (G-Rg1) was studied using fluorescence, ultraviolet–visible (UV–vis), and circular dichroism (CD) spectroscopies along with enzyme activity assay and molecular docking. The enzyme activity assays showed that G-Rg1 inhibited the activity of trypsin effectively. The fluorescence experiments indicated that a complex of G-Rg1–trypsin was formed and that the fluorescence of trypsin was quenched by G-Rg1 via a mixed-quenching mechanism (both static and dynamic quenching). The thermodynamic analysis suggested that hydrophobic interaction and hydrogen bond were the major forces between G-Rg1 and trypsin. According to the theory of Förster’s non-radiation energy transfer, the binding distance between trypsin and G-Rg1 was calculated to be 2.01 nm, which implies that energy transfer occurred within the complex. The experimental results obtained from UV–vis absorption spectra, synchronous fluorescence spectra, and CD spectra indicated that G-Rg1 was mainly located on tryptophan moiety and that the interaction between G-Rg1 and trypsin led to conformational changes of trypsin with some α-helix and unordered coil structures being transformed into β-sheet structures. In addition, docking results supported the above experimental findings and suggested the possible binding location of G-Rg1 on trypsin along with the possible hydrogen bonds and hydrophobic interactions between G-Rg1 and trypsin. The experimental results from this study should be useful to minimize the antinutritional effects and make full use of Genseng extracts in the food industry and also be helpful to the design of the drugs for the diseases related to overexpression of trypsin.Communicated by Ramaswamy H. Sarma
研究领域:
生物化学
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药物设计
·
食品科学
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DOI
PubMed
高影响因子
ACS Applied Bio Materials
·
2018
·
IF 0
Qingqing Tan,Ruirui Zhang,Weisu Kong,Fengli Qu,Limin Lu
A novel “turn-on” fluorescent trypsin detection platform dependant on carbon dot-MnO2 (CD-MnO2) nanocomposites and ascorbic acid-loaded apoferritin (APOAA) was fabricated. The detection mechanism relied on trypsin-catalyzed enzymolysis of APOAA, which released ascorbic acid (AA) as a reducing agent to disintegrate the MnO2 nanosheets, causing the recovery of the fluorescence of CDs. An excellent performance and high sensitivity of trypsin determination were observed with a detection limit (LOD) of 0.3411 ng/mL. This work provides us with a unique strategy for trypsin detection in human serum samples, which reveals the potential applications in clinical detection.
研究领域:
生物化学
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医学诊断
·
纳米材料科学
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产品关联
DOI
PubMed
JOURNAL OF CHROMATOGRAPHY A
·
2015
·
IF 4.169
Yuan Xiao,Jialiang Guo,Danni Ran,Qianqian Duan,Jacques Crommen,Zhengjin Jiang
A facile and efficient “one-pot” copolymerization strategy was used for the preparation of sulfonamide drug (SA) functionalized monolithic columns. Two novel SA-immobilized methacrylate monolithic columns, i.e. poly(GMA-SMX- co -EDMA) and poly(GMA-SAA- co -EDMA) were prepared by one-pot in situ copolymerization of the drug ligand (sulfamethoxazole (SMX) or sulfanilamide (SAA)), the monomer (glycidyl methacrylate, GMA) and the cross-linker (ethylene dimethacrylate, EDMA) within 100 μm i.d. capillaries under optimized polymerization conditions. The physicochemical properties and column performance of the fabricated monolithic columns were characterized by elemental analysis, scanning electron microscopy and micro-HPLC. Satisfactory column permeability, efficiency and separation performance were obtained on the optimized poly(GMA-SMX- co -EDMA) monolithic column for small molecules, such as a standard test mixture and eight aromatic ketones. Notably, it was found that the poly(GMA-SMX- co -EDMA) monolith showed a selective affinity to trypsin, while the poly(GMA-SAA- co -EDMA) monolith containing sulfanilamide did not exhibit such affinity at all. This research not only provides a novel monolith for the selective isolation and purification of trypsin, but it also offers the possibility to easily prepare novel drug functionalized methacrylate monoliths through a one-pot copolymerization strategy.
研究领域:
分析化学
·
生物化学
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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）