JOURNAL OF MOLECULAR LIQUIDS
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2022
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IF 6.633
Yuanyuan Yue,Qimin Tu,Yiying Guo,Yunting Wang,Yue Xu,Yilin Zhang,Jianming Liu
In this work, the binding affinity of fanetizole with proteases pepsin and trypsin were investigated by spectroscopy and molecular docking. As a thiazole derivative, the interactions of fanetizole with either pepsin or trypsin resulted in a fluorescence quenching. As such, the calculated values of K manifested that the fanetizole-pepsin system has a stronger binding affinity than fanetizole-trypsin at 296 K. Moreover, steady-state, time-resolved fluorescence and thermodynamic studies of the interaction of fanetizole with pepsin or trypsin suggested that a static fluorescence quenching mechanism and the interactions were primarily attributed to hydrogen-bonding interaction. The observed changes in synchronous fluorescence, circular dichroism spectroscopy, and FTIR results were related to the secondary construction alternations of both proteases. In line with fluorescence spectroscopy, molecular docking demonstrated similar binding characteristics and domains. The range of r values showed energy transfer from pepsin/trypsin to fanetizole.
研究领域:
生物化学
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药理学
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分子生物学
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DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2014
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IF 3.107
Yaping Wang,Guowen Zhang,Langhong Wang
Dimethyl phthalate (DMP) is widely used as a plasticizer in industrial processes and has been reported to possess potential toxicity to the human body. In this study, the interaction between DMP and trypsin in vitro was investigated. The results of fluorescence, UV–vis, circular dichroism, and Fourier transform infrared spectra along with cyclic voltammetric measurements indicated that the remarkable fluorescence quenching and conformational changes of trypsin resulted from the formation of a DMP–trypsin complex, which was driven mainly by hydrophobic interactions. The molecular docking and trypsin activity assay showed that DMP primarily interacted with the catalytic triad of trypsin and led to the inhibition of trypsin activity. The dimensions of the individual trypsin molecules were found to become larger after binding with DMP by atomic force microscopy imaging. This study offers a comprehensive picture of DMP–trypsin interaction, which is expected to provide insights into the toxicological effect of DMP.
研究领域:
生物化学
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毒理学
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环境科学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL BIOCHEMISTRY
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2015
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IF 2.219
Xiuxia Sun,Xiangdong Cai,Ren-Qi Wang,Jianxi Xiao
The preparation of biocatalysts based on immobilized trypsin is of great importance for both proteomic research and industrial applications. Here, we have developed a facile method to immobilize trypsin on hydrophobic cellulose-coated silica nanoparticles by surface adsorption. The immobilization conditions for the trypsin enzyme were optimized. The as-prepared biocatalyst was characterized by Fourier transform infrared spectroscopy , transmission electron microscopy, and elemental analysis . In comparison with free enzyme, the immobilized trypsin exhibited greater resistances against thermal inactivation and denaturants. In addition, the immobilized trypsin showed good durability for multiple recycling. The general applicability of the immobilized trypsin for proteomic studies was confirmed by enzymatic digestion of two widely used protein substrates: bovine serum albumin (BSA) and cytochrome c. The surface adsorption protocols for trypsin immobilization may provide a promising strategy for enzyme immobilization in general, with great potential for a range of applications in proteomic studies.
研究领域:
生物化学
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生物技术
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纳米技术
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蛋白质组学
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2021
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IF 10.618
Jiantao Ping,Wenli Wu,Lubin Qi,Jie Liu,Jinpeng Liu,Binglu Zhao,Quanbo Wang,Li Yu,Jin-Ming Lin,Qiongzheng Hu
The detection of trypsin and its inhibitor is significantly important for both clinical diagnosis and disease treatment. Herein, we demonstrate a hydrogel-assisted paper-based lateral flow sensor for the detection of trypsin and its inhibitor for the first time. The gelatin hydrogel is hydrolyzed based on the gel-to-sol transition in the presence of trypsin, which results in the release of the trapped water molecules in the gelatin hydrogel. By placing one end of a pH indicator strip onto the hydrolyzed gelatin hydrogel, water is flowing along the pH indicator strip. However, in the absence of trypsin, water cannot flow along the pH indicator strip as the water molecules are trapped in the gelatin hydrogel. The detection limit of the system reaches as low as 1.0 × 10 −6 mg/mL, and it is also applied to the quantitative detection of trypsin in human serum. In addition, the detection of a clinical drug aprotinin that is an inhibitor of trypsin is also successfully achieved. Noteworthy, only the gelatin hydrogel, pH indicator strip, and PS substrate are needed to fulfill the detection of trypsin without the need of other chemicals or reagents. Overall, we develop a particularly simple, elegant, robust, competitive, high-throughput, and low-cost approach for the rapid and label-free detection of trypsin and its inhibitor, which is very promising in the development of commercial products for sensing, diagnostic, and pharmaceutical applications . Besides, the hydrogel-assisted paper-based lateral flow sensor can also be employed to detect other analytes of interest by use of different stimuli-responsive hydrogel systems.
研究领域:
生物传感器
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临床诊断
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药物科学
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分析化学
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产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
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2022
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IF 9.231
Rui Cui,Shengnan Ji,Minquan Xia,Xing Fu,Xi Huang
Ovomucoid (OVM) is a critical anti-nutritional factor in egg, which may reduce nutrient utilization. In this study, the effects of polyphenols on the trypsin inhibitory activity (TIA) of OVM were investigated by exploring the structural changes and interaction mechanisms. The results found that TIA decreased to 62.34% and 90.41% as epigallocatechin gallate (EGCG) and gallic acid (GA) were added individually. EGCG and GA interacted with OVM via static quenching and hydrophobic interaction. They induced a transition of OVM conformation from disorder to order. Infrared and fluorescence quenching analysis showed that the interaction between EGCG or GA and OVM was spontaneous, and hydrophobic interaction was the predominant force. The mechanism suggested that polyphenols affect the protein conformation by spontaneously binding to OVM in hydrophobic interactions, and lowering the TIA through reduced hydrophobicity. In summary, EGCG may be a promising OVM trypsin activity inactivator, which could also guarantee safety of egg products.
研究领域:
食品科学
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生物化学
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营养学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
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2022
·Vol. 23
·期 19
·11853
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IF 6.208
Zichao Wang,Pei Liu,Ziyi Fang,He Jiang
Immobilized enzymes are a significant technological approach to retain enzyme activity and reduce enzyme catalytic cost. In this work, trypsin-incorporated Zn3(PO4)2hybrid nanoflowers were prepared via mild precipitation and coordination reactions. The controllable preparation of hybrid nanoflowers was achieved by systematically investigating the effects of the raw-material ratio, material concentration and reaction temperature on product morphology and physicochemical properties. The enzyme content of hybrid nanoflowers was about 6.5%, and the maximum specific surface area reached 68.35 m2/g. The hybrid nanoflowers exhibit excellent catalytic activity and environmental tolerance compared to free trypsin, which was attributed to the orderly accumulation of nanosheets and proper anchoring formation. Further, the enzyme activity retention rate was still higher than 80% after 12 repeated uses. Therefore, trypsin/Zn3(PO4)2hybrid nanoflowers—which combine functionalities of excellent heat resistance, storage stability and reusability—exhibit potential industrial application prospects.Keywords:hybrid materials;trypsin;nanoflowers;controlled synthesis;catalytic performance
研究领域:
生物化学
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材料科学
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产品关联
DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
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2020
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IF 3.232
Yanqing Wang,Qianqian Han,Gencheng Zhang,Hongmei Zhang
In this work, the effects of I 2 on the activities and conformational structures of digestive enzymes, trypsin and pepsin were studied. The results indicated that the enzyme activities were decreased to some extent in the presence of I 2 , especially trypsin. Upon gradual addition of I 2 , the intrinsic fluorescence quenching of trypsin and pepsin were observed by mainly static collision and hydrophobic forces . I 2 is more likely to cause the fluorescence quenching of trypsin than that of pepsin. Compared with pepsin, trypsin has a greater ability to bind with I 2 . The synchronous fluorescence spectral results indicated that I 2 induced the quaternary structure changes of trypsin/pepsin and changed the hydrophobicity of Tyr and Trp residues . In addition, molecular docking was used to obtain the binding mode and the various amino acid residues of trypsin and pepsin with I 2 . These investigations may constitute a solid work to further explain the process of migration and transformation of I 2 in digestive system.
研究领域:
生物化学
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分子生物学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2026
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IF 8.7
Xiao-Jing Wang,Yan Liang,Chen-Ying Wang,Ying-Qiu Li,Xiang-Zhong Zhao,Ting Wang
Synergistic hydrolysis of Ela-Try yields EIPs with 54.17% elastase inhibition. • Hydrolysis kinetics follow either pseudo-first-order or pseudo-second-order. • Elastase acts as the pioneer and key participant, with trypsin as collaborator. • Cytochrome C is identified as a key protein in the formation of highly active EIPs. • Mechanistic analysis showed hydrolysis follows stepwise synergistic mechanism. Due to the incomplete understanding of enzymatic degradation pathways of complex marine proteins, the production of highly active elastase inhibitory peptides (EIPs) remains limited by low yield, unstable activity, and poor controllability. Therefore, in this study, skipjack tuna cardiac arterial bulbs was used as the substrate, and kinetic analysis of pancreatic enzyme-mediated hydrolysis was performed to clarify the hydrolysis mechanism and obtain highly active EIPs. LC-MS/MS analysis identified 17 parent proteins and 430 unique peptides, with myoglobin, cytochrome C and elastin as the main bioactive precursors. The synergistic action of elastase and trypsin (Ela-Try) enhanced EIPs release, achieving 54.17% elastase inhibitory activity at the optimal enzyme ratio (elastase:trypsin = 1.2:1). Enzyme kinetics, molecular docking, and molecular dynamics simulations show that Ela-Try has lower K m (2.3751 mg/mL) and higher V max (0.0199 mg/mL·min) than elastase or trypsin alone, demonstrating synergistic hydrolysis. Ela-Try performs stepwise hydrolysis of proteins (especially cytochrome C), with elastase first cleaving hydrophobic residues (Gly, Ala, Val) according to a pseudo-first-order kinetic model, causing conformational changes and exposing new active sites (Lys, Arg); subsequently, trypsin cooperates with elastase following a pseudo-second-order kinetic model to further cleave the newly exposed sites and remaining hydrophobic residues. This study revealed the hydrolysis mechanism of the cooperative formation of highly active EIPs by Ela-Try, improving the release efficiency, activity and controllability of EIPs, and providing a theoretical basis for the high-value utilization of marine protein resources and the development of functional peptides. Download: Download high-res image (150KB) Download: Download full-size image
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DOI
PubMed
高影响因子
INDUSTRIAL CROPS AND PRODUCTS
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2021
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IF 5.645
Huan Wang,Chao Lei,Yunhang Zeng,Ying Song,Qixian Zhang,Bi Shi
Trypsin is used to remove non-collagenous proteins from cattle hide in leather processing but prone to damage hide surface and reduce leather quality due to its rapid protein hydrolysis action on the hide surface and slow penetration through the whole hide. A novel trypsin bating method was developed using a reversible inhibition of trypsin with high-yield, low-cost and environment-friendly soybean flour (SF). When the mass ratio of trypsin to SF was 1:20, the trypsin activity was decreased by 77 % at a reaction time of 5 min and completely returned in 100 min because of the competitive Kunitz trypsin inhibitor in SF. Bating with trypsin-SF mixture avoided the damage to hide surface caused by trypsin, improved the dispersion uniformity of hide collagen fibers and increased the softness and the mechanical strength of leather. This work exhibits huge potential for improving the leather quality and provides new insights into the soybean application.
研究领域:
生物化学
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皮革加工
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环境科学
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产品关联
DOI
高影响因子
RSC Advances
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2020
·Vol. 10
·期 45
·26765-26770
·
IF 3.119
Yiping Chen,Zuan Lin,Chenfang Miao,Qianqian Cai,Fenglan Li,Zongfu Zheng,Xinhua Lin,Yanjie Zheng,Shaohuang Weng
The development of a simple detection strategy for trypsin (Try) is urgent, and is ascribed to the diagnostic value of Try in several diseases. Herein, a facile but effective fluorescence strategy for Try was developed based on the protamine (Pro)-induced aggregation of carbon quantum dots (CQDs). The fluorescence of negatively charged CQDs was quenched with Pro due to the assembly of CQDs and Pro (CQDs/Pro) through electrostatic interaction. However, the highly positively charged Pro, which is rich in basic arginine residues, was preferred to be hydrolyzed by Try. Try can induce the deaggregation of CQDs/Pro, thereby enabling the release of CQDs to restore the fluorescence intensity. Thus, the use of CQDs/Pro as a testing platform will be employed as a “turn-on” method for Try. In addition, the fluorescence-resuming response was proportional to Try, ranging from 25 ng mL−1 to 500 ng mL−1 with a limit of detection (LOD) of 8.08 ng mL−1. This “turn-on” fluorescence assay for Try was label-free, convenient, and relatively free of interference from coexisting substances. Actual applications for Try monitoring and trypsin inhibitor screening also illustrated the considerable prospect of CQDs in the clinical field, combined with the superiority of the simple mixing operation.
研究领域:
生物化学
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医学诊断
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纳米材料
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产品关联
DOI
PubMed
高影响因子
RSC Advances
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2020
·Vol. 10
·期 35
·20862-20871
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IF 3.119
Guoyan Ren,He Sun,Gen Li,Jinling Fan,Lin Du,Guoting Cui
The molecular mechanism of interaction between aloe-emodin (AE) and trypsin was investigated, exhibiting remarkable outcomes. To detect the interaction mechanism, the binding of AE with trypsin was examined by a multi-spectroscopy and molecular docking method. Results showed that the binding of AE and trypsin would lead to static quenching and their binding forces were van der Waals forces and hydrogen bonding. The results of simultaneous and three-dimensional fluorescence spectroscopy showed that the combination of AE and trypsin caused changes in the microenvironment around the trypsin fluorophore, which might change the spatial structure of trypsin. FT-IR spectroscopy showed that the contents of α-helix and β-turn in trypsin were decreased and the contents of β-sheet, random coil and antiparallel β-sheet were increased. Moreover, all these experimental results were verified and reasonably explained by molecular docking results. We also investigated the enzyme activity of trypsin and the antioxidant activity of AE. The results showed that both the enzyme activity of trypsin and the antioxidant activity of AE were decreased after interaction between AE and trypsin. The findings outlined in this study should elucidate the molecular mechanisms of interaction between AE and trypsin and contribute to making full use of AE in the food industry.
研究领域:
生物化学
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食品科学
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药理学
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产品关联
DOI
PubMed
高影响因子
SOLID STATE SCIENCES
·
2018
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IF 1.861
Jianyi Liu,Yan Liu,Dianxin Jin,Minjia Meng,Yinhua Jiang,Liang Ni,Zhanchao Liu
Tannic-acid-templated large pore mesoporous silica nanoparticle (LMSN) was synthesized using one-pot procedure and the external surface was further modified with APTES (LMSN-N) to immobilize trypsin. After modification, trypsin was difficult to be immobilized on the outer surface of the LMSN due to the electrostatic repulsion. Naturally, study of the trypsin immobilized in the interior pores could be implemented solely. Meanwhile, the activity of immobilized trypsin was optimized by response surface method (RSM), and the ANOVA was employed to explore the interaction between independent variables. The maximum enzyme activity (12.28 U/mg) was obtained under a certain condition that the quality of T-LMSN-N-272 was 153 μg, the pH was 7.9, and the temperature was 40.4 °C. Simultaneously, the immobilized enzyme was proved to possess better thermal stability, storage stability and recycling performance than free enzyme.
研究领域:
生物化学
·
材料科学
·
酶工程
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）