Food Science and Human Wellness
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2023
·
IF 7
Jing Zhang,Di Wu,Lan Tang,Xia Hu,Zhen Zeng,Wen Wu,Fang Geng,Hui Li
Phlorizin (PHL) is a natural compound with strong antioxidant properties mainly found in apples. In this paper, the interaction mechanism of PHL with pepsin and trypsin was comparatively evaluated by computer simulation, fluorescence spectra, circular dichroism (CD), and fourier transform infrared (FT-IR) spectra at a molecular level. Fluorescence spectra showed that PHL quenches the pepsin/trypsin by static quenching. Thermodynamic parameters indicated that PHL binds to pepsin mainly through hydrogen bonds and van der Waals forces, and that of trypsin was electrostatic forces. The ground state complexes PHL and protease have a moderate affinity of 10 5 L/mol PHL binds more strongly to trypsin than to pepsin. CD and FT-IR spectra results showed that pepsin/trypsin decreased the β -sheet content and slightly changed its secondary structure upon PHL. These experimental results are mutually verified with the predicted computer-aid simulation results. Upon PHL and trypsin binding, the antioxidant capacity of PHL was elevated. Nevertheless, the antioxidant capacity of PHL was decreased after binding to pepsin. This work elucidates the binding of PHL binding mechanisms to pepsin/trypsin and provides useful information for the digestion of PHL to improve the application of PHL in food processing.
研究领域:
生物化学
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分子生物学
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食品科学
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高影响因子
ACS Applied Nano Materials
·
2025
·
IF 5.5
Junyao Li,Yujie Wang,Chen Jin,Zheng-Zhi Yin,Hongyu Zhang,Peiming Liu,Wenrong Cai,Datong Wu,Fengxiang Yin,Yong Kong
Trypsin (TRY) is an essential digestive enzyme, and its concentration level is regarded as a reliable indicator of pancreatic diseases. In this work, a highly sensitive and stable TRY electrochemical biosensor has been developed by taking advantage of the regulatable chiral recognition capability of gold nanoparticles/bovine serum albumin (AuNPs/BSA). Bovine serum albumin (BSA) is a protein with intrinsic chirality, which can be utilized to coat gold nanoparticles (AuNPs) through Au–S bonds and form AuNPs/BSA. Notably, the chiral recognition capability of AuNPs induced by d-Cysteine toward tyrosine (Tyr) enantiomers is opposite to that of BSA. For AuNPs/BSA, the chiral recognition capability is dominated by BSA adsorbed on the surface of AuNPs. By introducing varying concentrations of TRY, the structural integrity of BSA on the surface of AuNPs/BSA is gradually disrupted, resulting in a reversal of its chiral discrimination capability toward Tyr enantiomers, thereby enabling precise regulation of the chiral recognition capability of AuNPs/BSA relying on the specific hydrolysis of TRY. Therefore, the concentration of TRY can be sensitively detected by measuring the different recognition efficiencies of AuNPs/BSA/TRY toward Tyr enantiomers, namely, the peak current ratio of l-tyrosine to d-tyrosine (IL/ID) in differential pulse voltammetry. Additionally, the integration of dual electrochemical signals is conducive to enhancing the stability of the TRY detection results, manifesting the considerable application potential of the TRY electrochemical biosensor in biological analysis and medical diagnostics.
研究领域:
生物分析化学
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电化学传感器
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纳米生物技术
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医学诊断
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蛋白质工程
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COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS
·
2026
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IF 5.4
Yuqing Li,Hui Ye,Luping Su,Chuan Gao,Yuzhong Zhang,Hong Li
ZIF-8 was uniformly anchored onto the membrane pores, and then trypsin was immobilized via chelation. • Asymmetric interconnected pores enhance trypsin-substrate contact and reduce fouling. • The trypsin loading per unit mass of ZIF‑8 embedded in membranes was 1.58-fold that of ZIF‑8 crystals. • EVAL/ZIF-8-TR membrane exhibits superior affinity and reaction rate. • BSA degradation efficiency was significantly improved during filtration. Proteolysis plays an important role in various fields but suffers from several limitations, including autolysis, time consumption, low efficiency, and difficulty in enzyme recovery. Biocatalytic membranes integrate enzyme immobilization and membrane separation to enhance stability and reusability, but their application in proteolysis remains challenging due to unstable enzyme loading and high mass transfer resistance. Herein, biocatalytic membranes were fabricated by embedding zeolitic imidazolate framework‑8 (ZIF‑8) into ethylene vinyl alcohol copolymer (EVAL) via the non-solvent induced phase separation (NIPS) method, followed by trypsin incorporation. The ZIF‑8 nanocrystals were anchored on membrane pores, enhancing the stability of ZIF‑8 within the porous structure, preventing aggregation, and providing abundant adsorptive sites. The asymmetric structures of the membranes were obtained by NIPS, which was beneficial for reducing fouling and provided a large surface area for enzyme loading and enhanced substrate-enzyme contact. Trypsin was loaded onto ZIF‑8 through chelation, and the high porosity and connectivity of the membrane were retained. The trypsin loading reached 0.34 mg/cm², and the loading per unit mass of ZIF‑8 embedded in membranes was 1.58-fold that of ZIF‑8 crystals. Higher affinity and reaction rate of biocatalytic membranes during dynamic filtration were obtained when compared to free trypsin from Michaelis-Menten kinetic parameters. Moreover, pressure‑driven convective transport overcame diffusion limitations, significantly improving bovine serum albumin hydrolysis efficiency. This work holds great promise for enzyme-immobilized biocatalytic membranes, and offers a green and sustainable solution for high-efficiency proteolysis. Download: Download high-res image (178KB) Download: Download full-size image
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ANALYTICAL CHEMISTRY
·
2026
·
IF 7.3
Tongtong Zhai,Jing Li,Erkang Wang
Traditional molecular chemiluminescence (CL) often suffers from low photon yields because each event typically produces limited emission. Self-amplification using nanomaterial probes with high-density ligands can increase photon output per luminophore; however, widely used CdTe nanocrystals and perovskites remain constrained by poor stability and biocompatibility. Herein, we report a protein-engineered bovine serum albumin-stabilized gold nanocluster (BSA-Au NCs) CL probe that delivers high-intensity emission while addressing these limitations. The CL intensity depends on BSA concentration and the triggering oxidant, and structural analyses indicate that a more loosely folded secondary structure favors stronger CL. Notably, NaClO oxidizes the BSA ligand shell and provides sufficient energy to excite Au NCs, producing an ∼800-fold higher CL signal than H2O2 at the same concentration. The red-shifted CL spectrum relative to photoluminescence, together with X-ray photoelectron spectroscopy, supports a ligand-involved emission mechanism, which is further validated by BSA-specific recognition experiments. Leveraging ligand-shell disruption, we construct a signal-off CL assay for ultrasensitive trypsin detection with a linear range of 0.01–500 μg/mL and a detection limit of 3.3 ng/mL (S/N = 3). This work provides a robust trypsin sensing platform and mechanistic insight into ligand-involved CL in BSA-Au NCs, enabling the rational design of functional Au nanocluster-based luminescent materials.
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高影响因子
JOURNAL OF ENVIRONMENTAL SCIENCE AND HEALTH PART A-TOXIC/HAZARDOUS SUBSTANCES & ENVIRONMENTAL ENGINEERING
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2025
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IF 2.1
Zijie Sun,Yaoyao Wang,Rui Guan,Hongbin Liu,Jing Lan,Hui Xue,Wansong Zong
Triclocarban (TCC), a widely used antimicrobial agent, may threaten ecosystems and human health via bioaccumulation, necessitating study of its protein interactions to understand molecular toxicity. In this paper, trypsin (TRY) was utilized as a model protein to explore its binding to TRY. The results revealed that the binding could result in a reduction of the enzymatic activity of TRY. Spectra analysis showed that TCC could heighten the quenching effect on the intrinsic fluorescence of TRY. The fluorescence quenching of TRY encompassed dynamic and static quenching mechanisms. The association constants (Ka) exhibited a high magnitude (∼106) at both 293 and 313 K, indicating a robust affinity between the two entities. Molecular docking studies and thermodynamic parameters (ΔH < 0, ΔS < 0) suggested hydrogen bonds and van der Waals forces are necessary for TCC’s binding to TRY. The formation of the TRY-TCC complex induced alterations in the secondary structure and local microenvironment of TRY, leading to a more relaxed skeletal structure. This paper will provide a fundamental basis for further studying the molecular toxicity of TCC in living organisms. Future in vivo studies will be essential to establish the physiological consequences of TCC-TRY binding in biological systems.
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高影响因子
JOURNAL OF COLLOID AND INTERFACE SCIENCE
·
2025
·
IF 9.7
Qizhen Huang,Yulin Yang,Shufang Hong,Yuanyuan Wang,Yuanyuan Liu,Huiling Lan,Yun Wang,Juan Han,Lei Wang
A novel intelligent polymeric bioreactor was designed through the switch of core-shell structure, which exhibited the controllable shell permeability and enrichment effect, leading to an improvement in the hydrolysis activity. During the preparation process, the polymeric micelles generated a large amount of accessible binding sites on the soluble shell for trypsin immobilization. Subsequently, a core-shell exchange created a rigid protective cavity to stabilize the immobilized trypsin. The immobilization capacity reached 352 mg/g within 60 min (81 % immobilization efficiency), which was 7.6 times that of UCST-type polymers (PMAAc). 86 %, 82 %, 66 % of its original activity was maintained under strong acid (pH = 3), alkali (pH = 8), high temperature (55 °C) for 120 min, respectively. Furthermore, the controllable shell permeability and enrichment effect of the polymeric bioreactors facilitated the diffusion and enrichment of large molecule substrates into active sites for catalysis, giving 16 % increase in the degree of hydrolysis and 10 % improvement in antioxidant activity of almond protein.
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高影响因子
TALANTA
·
2024
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IF 5.6
Ya-Qi Zhao,Zhi-Peng Li,Shi-Chao Dong,Hao Wang,Yi-Mei Zhao,Lin-Yi Dong,Zhen-Yu Zhao,Xian-Hua Wang
The effective method for trypsin purification should be established because trypsin has important economic value. In this work, a novel and simple strategy was proposed for fabricating micron-sized magnetic Fe 3 O 4 @agarose-benzamidine beads (MABB) with benzamidine as a ligand, which can efficiently and selectively capture trypsin. The micro-sized MABB, with clear spherical core-shell structure and average particle size of 6.6 μm, showed excellent suspension ability and magnetic responsiveness in aqueous solution. The adsorption capacity and selectivity of MABB towards target trypsin were significantly better than those of non-target lysozyme. According to the Langmuir equation, the maximum adsorption capacity of MABB for trypsin was 1946 mg g −1 at 25 °C, and the adsorption should be a physical sorption process. Furthermore, the initial adsorption rate and half equilibrium time of MABB toward trypsin were 787.4 mg g −1 min −1 and 0.71 min, respectively. To prove the practicability, MABB-based magnetic solid-phase extraction (MSPE) was proposed, and the related parameters were optimized in detail to improve the purification efficiency. With Tris-HCl buffer (50 mM, 10 mM CaCl 2 , pH 8.0) as extraction buffer, Tris-HCl buffer (50 mM, 100 mM CaCl 2 , pH 8.0) as rinsing buffer, acidic eluent (0.01 M HCl, 0.5 M NaCl, pH 2.0) as eluent buffer and alkaline buffer (1 M Tris-HCl buffer, pH 10.0) as neutralization solution, the MABB-based MSPE was successfully used for trypsin purification from the viscera of grass carp ( Ctenopharyngodon idella ). The molecular weight of purified trypsin was determined as approximate 23 kDa through sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The purified trypsin was highly active from 30 °C to 60 °C, with an optimum temperature of 50 °C, and was tolerant to pH variation, exhibiting 85 % of maximum enzyme activity from pH 7.0 to 10.0. The results demonstrated that the proposed MABB-based MSPE could effectively purify trypsin and ensure the biological activity of purified trypsin. Therefore, we believe that the novel MABB could be applicable for efficient purification of trypsin from complex biological systems.
研究领域:
生物化学
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生物技术
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分析化学
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蛋白质纯化
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酶学
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PubMed
高影响因子
MACROMOLECULAR RAPID COMMUNICATIONS
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2014
·Vol. 35
·期 18
·1606-1610
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IF 4.608
Yan Lv,Jianbin Zhang,Yizhe Song,Bing Wang,Shujun Wang,Shan Zhao,Guojun Lv,Xiaojun Ma
Sodium alginate (SA), acting as a trypsin inhibitor by means of electrostatic interaction, is studied. The half-maximal inhibitory concentration (IC 50 = 0.05 μg mL −1 ) of this natural anionic polymer is about 400 times lower than that of commercial soybean trypsin inhibitor (STI). Unlike the Ca 2+ -deprivation mechanisms, its inhibition may be attributed to preventing the trypsin active site (TAS) from accessing the macromolecular substrates instead of denaturing it. SA is an efficient, innocuous, and cost-effective inhibitory excipient that can be conveniently used in many peptide and protein dosage formulations.
研究领域:
生物化学
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药剂学
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高分子科学
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酶抑制
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高影响因子
Journal of Chemistry
·
2020
·
IF 1.79
Di An,Liang Li
Nanofibril system constructed by protein self-assembly is widely used in the food industry because of purposive functional properties. Soy protein isolate nanofibrils (SPINs) were reported to form via heating at pH 2.0. In this research, the soy protein isolate (SPI) hydrolysate prepared by trypsin was used as a raw material for the formation of nanofibrils called soy protein isolate hydrolysate nanofibrils (SPIHNs). Microscopic images demonstrated the formation of two nanofibrils. Based on circular dichroism spectroscopy and Thioflavin T (ThT) fluorescence spectral, we concluded that β-sheet played an important role in SPIN and SPIHN’s structural composition. At the same time, the α-helix in SPI had not been destroyed, thereby favoring the formation of SPIHN. The surface hydrophobicity of SPIHN continued to increase during the heating process and reached the highest value when heating 8 h. SDS-PAGE analysis showed that peptides produced by enzyme-modified SPI affected the formation of SPIHN. These results proposed that enzymatic hydrolysis prior to acidic during fibrillation process affected the fibrillation of SPI, and the peptides formed by enzymatic hydrolysis were more efficient for the self-assembly process. This study will provide a theoretical basis for the future research of SPI nanofibril functionality.
研究领域:
食品科学
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蛋白质化学
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纳米技术
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DOI
高影响因子
Materials Science and Engineering B-Advanced Functional Solid-State Materials
·
2021
·
IF 4.051
Xianbao Shi,Ping Zhang,Lingqiong Wu,Xiangyu Zhao,Jinzhu Jiang,Jing Li,Bo Yang
Mesoporous silica synthesized using biomimetic method is superior for its mild synthesis process and source saving advantages. Herein, trypsin was used as template to synthesize biomimetic mesoporous silica, and we named it biomimetic large mesoporous silica (BL-MS) due to its large mesopores. For comparison, commonly used mesoporous silica nanoparticles (MSNs) were studied along with BL-MS. The study focused on their different characteristics and abilities in delivering poorly water-soluble drug indomethacin (IMC). It turned out that BL-MS were nanoparticles with large mesopores while MSNs were nanospheres with small mesopores. As for their application, BL-MS had stronger energy to adsorb drug molecules, which can be one of its advantages as drug carrier. BL-MS and MSNs improved IMC dissolution for almost 3.5 times and 2.7 times respectively. Furthermore, BL-MS performed superior oral bioavailability and anti-inflammatory effect, which confirmed the outstanding advantages of BL-MS in delivering poorly water-soluble drugs due to its large mesopores.
研究领域:
材料科学
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药学
·
生物医学工程
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高影响因子
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
·
2020
·
IF 3.391
Jia-Yu Shi,Hai-Yan Pan,Kun Liu,Min Pan,Guo-Jun Si
Background Our previous research revealed that trypsin is abundantly expressed in atherosclerotic plaques and its distribution overlaps with that of matrix metalloproteinase-9 (MMP-9). This study was performed to explore the possible roles of trypsin in vulnerable atherosclerotic plaque formation. Methods and results Twenty-four rabbits were randomly assigned to a normal (control) group, an atherosclerosis (experimental) group and a trypsin inhibitor (aprotinin) group. In the 13th feeding week, the aprotinin group was treated with 5 mg/kg/day aprotinin via ear vein for 4 weeks. At the end of the 16th week, coronary arterial and aortic expression of trypsin, proteinase-activated receptor-2 (PAR-2), activated MMP-9, and pro-inflammatory cytokines were significantly greater in the experimental group than in the control group. Aprotinin decreased trypsin expression and activation in plaques, blocked PAR-2 and MMP-9 activation, and decreased cytokine expression; it also increased fibrous cap thickness, decreased the intima-media thickness and intimal/medial ratio, thus significantly ameliorating plaque vulnerability. Upregulated trypsin, MMP-9 and PAR-2 were also found in coronary intimal atherosclerotic plaques of patients undergoing coronary artery bypass grafting. Conclusions Ectopic trypsin was significantly upregulated in atherosclerotic plaques, which increased pro-inflammatory cytokine levels by activating PAR-2 and promoted plaque instability by activating proMMP-9, thereby promoting atherosclerosis and plaque vulnerability. In addition, the high trypsin expression in human coronary intimal atherosclerotic plaques suggests that targeting trypsin may be a new strategy for acute coronary syndrome prevention.
研究领域:
心脏病学
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药理学
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分子生物学
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DOI
PubMed
高影响因子
Innovative Food Science & Emerging Technologies
·
2012
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IF 3.03
Wei Liu,Zhao-Qin Zhang,Cheng-Mei Liu,Ming-Yong Xie,Rui-Hong Liang,Jun-Ping Liu,Li-Qiang Zou,Jie Wan
Previous research indicated that the unfolding of trypsin induced by dynamic high–pressure microfluidization (DHPM) treatment had enhanced its hydrolytic stability. However, the unfolding trypsin is in a metastable state and will subsequently aggregate to be in the lowest energy state. In this study, monomethoxy polyethylene glycol-succinimidyl carbonates (mPEG–SC) was chosen to modify trypsin acted as a molecular patch. The results indicated mPEG–SC showed a significant effect on storage stability and thermal stability of unfolded trypsin. Compared to trypsin DHPM–treated under pressure of 100 MPa, further mPEG–SC modification increased the activity from 50% to 74% and from 70% to 87% after 4 °C for 8 days and 55 °C for 10 min, respectively. Fluorescence quenching demonstrated the conjugation of mPEG–SC and trypsin, and indicated molecular patch modification could inhibit the refolding of unfolded trypsin. Industrial relevance Dynamic high–pressure microfluidization (DHPM) is a promising technology that has been widely applied for innovative foods with modified functional properties or structure. Trypsin has been widely used in modern food industry and biological field. Either DHPM treatment or mPEG–SC modification enhanced the thermal stability of trypsin, while, the combination of the two enhanced the thermal and storage stability of trypsin more significantly. It is expected that this study can provide a new viewpoint and technical scheme for basic research on theory and application of enzyme modification .
研究领域:
食品科学
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生物化学
·
酶工程
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高影响因子
危险品化学品经营许可证（带存储）