TALANTA
·
2017
·
IF 4.162
Mingmin Wu,Xiaoyong Wang,Kun Wang,Zijian Guo
Trypsin and its inhibitors are relevant to many physiological processes and diseases. In this study, a nanosensor capable of detecting trypsin and its inhibitors was designed based on the fluorescence resonance energy transfer (FRET) between upconversion nanoparticle (UCNP) and gold nanoparticle (AuNP). UCNP and AuNP were linked by a trypsin-sensitive peptide DDDDARC, forming the non-fluorescent UCNP-peptide-AuNP nanosensor. In the presence of trypsin, the peptide was cleaved and the quenched fluorescence was restored; in the presence of trypsin inhibitors , the recovery of the fluorescence was decreased. The nanosensor showed a superb sensitivity and selectivity for trypsin and its inhibitors, with a detection limit of 4.15 ng mL −1 for trypsin. UCNP-peptide-AuNP could eliminate the interference of background fluorescence and avoid the light toxicity, and potentially be used to diagnose trypsin-related diseases or screen trypsin inhibitors.
研究领域:
生物化学
·
纳米医学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD RESEARCH INTERNATIONAL
·
2015
·
IF 2.818
Huizhi Xiao,Benguo Liu,Haizhen Mo,Guizhao Liang
In this work, the inhibitory effects of tannic acid on the α-glucosidase and trypsin were systematically evaluated by comparing with the clinical diabetes healer acarbose and the soybean-derived trypsin inhibitor using fluorescence spectroscopy and enzymatic kinetics methods. We showed that the anti-α-glucosidase activity of tannic acid (IC 50 = 0.44 μg/mL) was higher than that of acarbose (IC 50 > 0.60 μg/mL), while its anti-trypsin activity (IC 50 = 0.79 mg/mL) was significantly lower than that of the trypsin inhibitor from soybean (IC 50 < 0.20 mg/mL). Enzymatic kinetics measurements confirmed that the inhibitory pattern of tannic acid toward two tested enzymes was a mixed competitive and noncompetitive inhibition. Tannic acid could bind the enzymes to form new complexes, presenting a strong static fluorescence quenching. The presence of tannic acid led to the hypsochromic shift of the maximum fluorescence in trypsin, but not in α-glucosidase. The thermodynamic parameters indicated that the main driving force between tannic acid and both the enzymes was the hydrophobic interaction followed by the electrostatic interaction. Our work suggests that tannic acid is a strong anti-α-glucosidase natural inhibitor with a low inhibitory activity for trypsin, thus its roles in functional food and medicinal plants should be re-recognized.
研究领域:
食品科学
·
生物化学
·
药理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF MOLECULAR STRUCTURE
·
2015
·
IF 1.602
Yan-Qing Wang,Hong-Mei Zhang,Yi-Jun Kang,Yun-Lan Gu,Jian Cao
In the present study, the experimental and theoretical methods were used to analyze the binding interaction of food dye , curcumin with trypsin. The results of fluorescence spectroscopic measurements indicated that curcumin binding resulted in the obviously intrinsic fluorescence quenching with the increase concentration of curcumin. This binding interaction is a spontaneous process with the estimated enthalpy and entropy changes being −15.70 kJ mol −1 and 40.25 J mol −1 K −1 , respectively. Hydrogen bonds and hydrophobic forces played an important role in the complex formation between curcumin and trypsin. Moreover, curcumin could enter into the primary substrate-binding pocket and makes the activity of trypsin decrease remarkably with the increasing concentration of curcumin.
研究领域:
生物化学
·
分子生物学
·
药物化学
✓ 已验证
产品关联
DOI
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2013
·
IF 1.977
Wu He,Huanjing Dou,Lu Zhang,Lvjing Wang,Ruiyong Wang,Junbiao Chang
The interactions between Trypsin and Bicyclol or analogs (Bifendate, I, II and III) were investigated by spectrophotometric methods. It was found that Bicyclol or analogs had strong ability to quench the intrinsic fluorescence of Trypsin by a static quenching procedure. The binding constants were obtained at three temperatures. The thermodynamics parameters reveal that the hydrophobic and electrostatic interactions play an important role in the interaction. Results showed that the microenvironments of tryptophan residue of Trypsin were disturbed by the analogs. Results indicated that Bifendate was the strongest quencher among five compounds.
研究领域:
生物化学
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药物化学
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分子生物学
·
光谱学
✓ 已验证
DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2015
·
IF 4.513
Juan Qiao,Jin Yong Kim,Yuan Yuan Wang,Li Qi,Fu Yi Wang,Myeong Hee Moon
Fast and effective protein digestion is a vital process for mass spectrometry (MS) based protein analysis . This study introduces a porous polymer membrane enzyme reactor (PPMER) coupled to nanoflow liquid chromatography-tandem MS (nLC-ESI-MS/MS) for on-line digestion and analysis of proteins. Poly (styrene- co -maleic anhydride) (PS- co -MAn) was fabricated by the breath figure method to make a porous polymer membrane in which the MAn group was covalently bound to enzyme. Based on this strategy, microscale PPMER (μPPMER) was constructed for on-line connection with the nLC-ESI-MS/MS system. Its capability for enzymatic digestion with bovine serum albumin (BSA) was evaluated with varied digestion periods. The on-line proteolysis of BSA and subsequent analysis with μPPMER-nLC-ESI-MS/MS revealed that peptide sequence coverage increased from 10.3% (digestion time 10 min) to 89.1% (digestion time 30 min). μPPMER can efficiently digest proteins due to the microscopic confinement effect, showing its potential application in fast protein identification and protease immobilization. Applications of on-line digestion using μPPMER with human plasma and urinary proteome samples showed that the developed on-line method yielded equivalent or better performance in protein coverage and identified more membrane proteins than the in-solution method. This may be due to easy accommodation of hydrophobic membrane proteins within membrane pores .
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
FOOD CHEMISTRY
·
2017
·
IF 4.529
Benguo Liu,Huizhi Xiao,Jiaqi Li,Sheng Geng,Hanjun Ma,Guizhao Liang
Trypsin is a kind of protease for digestion and food processing, whose activity can be inhibited by phenolic acids in plant foods. However, most reports explained the inhibitory difference of phenolic acids based on the number and position of substituent groups, which failed to reveal the comprehensive inhibitory mechanism. In this work, the inhibitory effects of 11 common phenolic acids on trypsin were investigated. Amongst the tested cinnamic and benzoic acid derivatives, caffeic acid and gallic acid showed the strongest anti-trypsin activity with a noncompetitive inhibition pattern, respectively. The fluorescence analysis displayed that both the quenching rate constant (K q ) and binding constant (K A ) of caffeic acid were higher than those of gallic acid. Molecular docking illustrated their different binding modes with trypsin. The ONIOM calculations revealed that the binding capacity of caffeic acid was higher than that of gallic acid, which could explain their difference in their inhibitory behaviors.
研究领域:
食品科学
·
生物化学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2017
·
IF 4.529
Junguang Li,Qisen Xiang,Xiufang Liu,Tian Ding,Xiangsheng Zhang,Yafei Zhai,Yanhong Bai
Soybean trypsin inhibitor (STI) is considered as one of the most important anti-nutritional factors in soybeans. The objective of this study was to investigate the impacts and underling mechanisms of dielectric-barrier discharge (DBD) plasma on STI activities. The results shown that DBD plasma treatment significantly induced the inactivation of STI in soymilk and Kunitz-type trypsin inhibitor from soybean (SKTI) in a model system. After exposure to DBD plasma at 51.4 W for 21 min, the STI activities of soymilk were reduced by 86.1%. Affter being treated by DBD plasma, the intrinsic fluorescence and surface hydrophobicity of SKTI were significantly decreased, while the sulfhydryl contents were increased. It is assumed that DBD plasma-induced conformational changes and oxidative modification might contribute to the inactivation of SKTI. In summary, DBD plasma technology is a potential alternative to heat treatment for the inactivation of anti-nutritional substances in food legumes.
研究领域:
食品科学
·
生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
PLANT GROWTH REGULATION
·
2023
·Vol. 100
·期 3
·633-646
·
IF 4.2
Wang Jie,Jia Jingyu,Sun Jiaju,Pang Xinyue,Li Bairu,Yuan Jiangfeng,Chen Enyan,Li Xin
Trypsin can significantly improve the storage quality of Cucumis sativus ( C. sativus ) due to its novel superoxide scavenging activity. Based on the results of physiological indices, an integrated analysis of widely targeted metabolomics and transcriptomics was carried out, gene function was further confirmed by VIGS, a potential molecular mechanism of resistance improvement of C. sativus due to flavonoid biosynthesis induced by trypsin was proposed and discussed. Transcriptomic results showed that 1,068 genes highly related to flavonoid biosynthesis gathered in the brown module were obtained by the advanced method WGCNA. An isomeric compound of dihydrochalcone, trilobatin, was likely to be the key metabolite regulated by trypsin. Results of O2PLS and co-expression network illustrated that CsUGT91C1 acted as the hub gene which regulated the biosynthesis of flavonoids, especially trilobatin, in C. sativus . CsUGT91C1 also played as a hub node in the Protein-Protein Interactions network of WGCNA screened genes (WSGs). GO and KEGG analyses of WSGs enriched the transferase activities, especially UDP-glycosyltransferase activity and KEGG pathways of “Phenylpropanoid biosynthesis (map00940)” and “Flavonoid biosynthesis (map00941)”. Results of webtools of the PlantCARE database identified the G-box region of CsUGT91C1 , which can bind to the transcription factor CsMYC2. The expression and biological effects of CsUGT91C1 were significantly impeded by the silencing of CsMYC2 through VIGS. The results of this study might give new insight into the exploration of the postharvest resistance mechanism of C. sativus induced by trypsin and provide useful information for the subsequent mining of resistance genes in C. sativus . Graphical abstract
研究领域:
植物科学
·
分子生物学
·
生物化学
·
基因组学
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF THE CHINESE CHEMICAL SOCIETY
·
2026
·
IF 2
Jing Chen,Xiyan He,Yuzhi Wang,Xueqin Ding
Five deep eutectic solvent (DES)-functionalized magnetic polydopamine (MPDA) composites were designed and synthesized via an ultrasound-assisted method, and subsequently integrated with magnetic solid-phase adsorption (MSPA) for the efficient adsorption of trypsin. Among them, MPDA@[ChCl][EG], functionalized with a DES composed of choline chloride and ethylene glycol, exhibited the highest adsorption efficiency. The morphology, magnetic responsiveness, and functional group distribution of MPDA@[ChCl][EG] were characterized by transmission electron microscopy, vibrating sample magnetometry, and fourier-transform infrared spectroscopy. Single-factor optimization experiments were conducted to investigate the key parameters affecting trypsin adsorption. Under optimal conditions, MPDA@[ChCl][EG] achieved an adsorption capacity of 338.99 mg·g −1 for trypsin, with an equilibrium time of 60 min. The adsorption behavior was well described by the Langmuir isotherm and pseudo-second-order kinetic models, indicating monolayer chemisorption. Furthermore, MPDA@[ChCl][EG] was successfully applied to the adsorption and separation of trypsin from a complex biological sample (bovine pancreas crude extract). This strategy of constructing magnetic adsorbents using green DESs and MPDA offers new possibilities for protein adsorption under mild conditions.
✓ 已验证
DOI
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
·
2026
·
IF 3.6
Chen Li,Yue Yu,Xin Liu,Xiaodong Cui,Yuru Guo,Jiao Li,Fang Li
● Recombinant BTI was immobilized on PDA-coated Fe₃O₄ nanoparticles. ● Fe₃O₄@PDA@BTI enabled one-step trypsin purification from porcine pancreatic homogenate. ● The maximum trypsin adsorption capacity was 13.78 ± 0.34 mg/g. A magnetic affinity adsorbent, Fe₃O₄@PDA@BTI, was prepared by immobilizing recombinant buckwheat trypsin inhibitor (BTI) on polydopamine (PDA)-coated Fe₃O₄ nanoparticles. TEM, FTIR, XRD, VSM, BTI-loading analysis, and retained inhibitory activity collectively supported the successful construction of the material. The adsorbent recovered trypsin from porcine pancreatic extract in a single batch purification step, with a maximum adsorption capacity of 13.78 ± 0.34 mg/g. Fe₃O₄@PDA@BTI retained more than 90% of its inhibitory activity at pH 4–10, more than 85% of its inhibitory activity after 3 h at 30–80 ℃, and approximately 94% after 30 d at 4 ℃. It also exhibited good stability against ethanol, dimethyl sulfoxide (DMSO), urea, and sodium dodecyl sulfate (SDS). However, the relative adsorption capacity decreased to 45.35 ± 0.19% after 10 reuse cycles, indicating that ligand coupling and regeneration require further optimization. These results demonstrate the feasibility of combining recombinant inhibitor recognition with magnetic recovery for trypsin purification.
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产品关联
DOI
高影响因子
EUROPEAN JOURNAL OF PHARMACOLOGY
·
2025
·
IF 4.7
Yi-ni Lu,Xiao-han Wang,Bo-cheng Xu,Yufen Zheng,Xiong Zhang,Yi Liu,Tian-yin Wang,Feng Yu
Septic cardiomyopathy (SCM) represents one of the life-threatening manifestations of sepsis, for which specific therapies are currently lacking. The mechanism of action of urinary trypsin inhibitor (UTI), widely employed in treating acute inflammatory disorders, on SCM has remained elusive. Here, we observed that UTI alleviated cardiac function, reduced myocardial injury, and improved cardiac permeability in a cecum ligation and puncture (CLP)-induced sepsis mouse model. In addition, UTI significantly decreased serum and myocardial angiopoietin 2 (ANG2) levels in CLP mice, which is a biomarker associated with sepsis prognosis. In vitro experiments utilizing mouse myocardial microvascular endothelial cells (MMVECs) were conducted to explore the molecular mechanisms underlying UTI-mediated regulation of ANG2 expression, secretion, and its impacts on endothelial permeability and neutrophil adhesion. UTI mitigated endothelial barrier dysfunction and reduced neutrophil adherence following thrombin stimulation and ANG2 overexpression. The experiments further demonstrated that UTI-induced inhibition of ANG2 expression was related to the blockade of forkhead box O1 (FOXO1) and phosphorylated signal transducer and activator of transcription 3 (p-STAT3) nuclear transcription and that UTI disrupted ANG2 exocytosis by reducing intracellular Ca 2+ concentration, a mechanism distinct from other serine protease inhibitors. In addition, UTI ameliorated barrier impairment and neutrophil adhesion in ANG2 overexpression cells by restoring tyrosine kinase receptor 2 Tie2)/protein kinase B (AKT) pathway phosphorylation. Docking predictions and surface plasmon resonance (SPR) further corroborated that UTI is strongly bound to ANG2. Altogether, our findings revealed a novel perspective wherein UTI functions as a multifaceted ANG2 inhibitor in SCM. Thus, ANG2 constitutes a promising therapeutic target for septic cardiomyopathy.
研究领域:
生物学
·
医学
·
药理学
·
细胞生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF MOLECULAR LIQUIDS
·
2018
·
IF 4.513
Yunchang Fan,Xin Wang,Jing Li,Lina Zhang,Lei Yang,Pengfu Gao,Zhulin Zhou
The kinetics of the inhibitory effects of ionic liquids (ILs) on the trypsin activity was useful for the practical applications, herein, kinetic information of the IL-trypsin interaction was systematically studied by infrared (IR) and ultraviolet-visible (UV–Vis) spectroscopies. Experimental results indicate that the presence of ILs makes the Michaelis constant ( K m ) of the enzyme reaction increase, meanwhile, the maximum rate ( V max ) keeps constant, suggesting that the inhibition of ILs on the trypsin activity belongs to the reversible-competitive type. IR and UV–Vis spectra show that the presence of ILs does not change the secondary structures of trypsin. After removing the ILs, the trypsin activity can be recovered; the activation energy keeps constant without and with the presence of ILs.
研究领域:
生物化学
·
酶学
✓ 已验证
DOI
高影响因子
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