MICROCHEMICAL JOURNAL
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2021
·
IF 4.821
Qiaozhen Bao,Dan Lin,Yaoran Gao,Lina Wu,Jinhua Fu,Khuslen Galaa,Xinhua Lin,Liqing Lin
Herein we develop a novel “off–on-off” fluorescent nanosensor for the ultra-sensitive detection of protamine and trypsin based on the as-prepared nitrogen and sulphur doped carbon dots (N,S-CDs) and gold nanoparticles (AuNPs). N,S-CDs were treated as fluorophore, while AuNPs were treated as fluorescence absorber. By experimental studies, the fluorescence quenching mechanism of N,S-CDs by AuNPs was inner-filter effect (IFE). Upon addition of protamine to the mixed aqueous of N,S-CDs/AuNPs, the electrostatic interaction and Au-N bond between protamine and AuNPs resulted in AuNPs aggregation, and the quenched fluorescence of N,S-CDs could be recovered. Protamine is rich in arginine and can be specifically hydrolyzed by trypsin, resulting in the deaggregation of AuNPs and further quenching the fluorescence of N,S-CDs. Thus, we can determine the concentration of protamine and trypsin by measuring fluorescence intensity of N,S-CDs. Under the optimized conditions, the fluorescence intensity showed a good linear relationship with protamine concentration of 20–260 ng/mL with the detection limit of 4.7 ng/mL. Meanwhile, quantitative determination of trypsin in a range from 10 to 140 ng/mL with the detection limit of 4.3 ng/mL can be achieved in this fluorescent nanosensor. This fluorescent nanosensor also showed excellent selectivity, and we successfully applied it for the detection of protamine and trypsin in human urine samples. Based on the above findings, we conclude that this novel “off–on-off” fluorescent nanosensor has great potential in clinical diagnosis.
研究领域:
分析化学
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纳米技术
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生物医学工程
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DOI
高影响因子
Food Chemistry-X
·
2023
·
IF 6.1
Xin Qi,He Liu,Yongfang Ren,Yihao Zhu,Qiulu Wang,Yanqing Zhang,Yushu Wu,Lixia Yuan,Hui Yan,Min Liu
The combined application of multiple natural polyphenols in functional foods may provide better health benefits. The binding of polyphenols with different structures to proteins will affect their respective functions. Spectroscopy and molecular docking were used to investigate the competitive binding of chlorogenic acid (CGA)/caffeic acid (CA) and gallic acid (GA) to trypsin. The effects of different molecular structures and the order of adding the three phenolic acids on the binding were assessed. The stability of trypsin and its docked complexes with CGA/CA/GA was evaluated by molecular dynamics simulation. The effects of the binding process on the activity and thermal stability of trypsin, as well as on the antioxidant activity and stability of CGA/CA/GA were explored. The competitive binding of CGA/CA and GA to trypsin affected their synergistic antioxidant effects. The results may provide a reference for the combined application of CGA/CA and GA in food and pharmaceutical fields.
研究领域:
食品科学
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药理学
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生物化学
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分子生物学
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DOI
PubMed
高影响因子
Journal of Functional Foods
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2024
·
IF 3.8
Xueping Wang,Jiaxing Wang,Yurou Chu,Jipeng Sun,Ru Song,Bin Zhang
This study used skipjack by-products to produce skipjack trypsin hydrolysate (STH), which was then processed through the Maillard reaction to create its Maillard product, STHMS3 (200–1000 Da). We evaluated STHMS3's protective effects against hyperuricemia-induced renal injury in mice. Utilizing in vivo experiments, network pharmacology, and molecular docking, we explored its protective mechanisms. STHMS3 significantly decreased serum uric acid, creatinine, blood urea nitrogen, and xanthine oxidase (XOD) levels, reduced oxidative stress, and enhanced antioxidant protein levels in kidneys. Network pharmacology analysis showed STHMS3's interaction with multiple targets and pathways, including apoptosis, NF-κB, and TNF signaling pathways, suggesting a multi-level interactive network mechanism. Molecular docking confirmed STHMS3's ability to directly inhibit XOD through hydrogen bond formation. This study highlights the therapeutic potential of food-derived peptides in managing hyperuricemia and protecting renal function.
研究领域:
食品科学
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医学
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分子生物学
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DOI
高影响因子
Nanoscale
·
2013
·Vol. 5
·期 13
·5905-5911
·
IF 6.233
Lingxin Chen,Xiuli Fu,Jinhua Li
In this work, a simple and sensitive surface-enhanced Raman scattering (SERS) strategy was developed for recognition and detection of trypsin, by using anti-aggregation of 4-mercaptopyridine (4-MPY)-functionalized silver nanoparticles (AgNPs) based on the interaction between protamine and trypsin. The polycationic protamine not only served as a substrate for enzyme hydrolysis but also worked as a medium for SERS enhancement, which could bind negatively charged 4-MPY-functionalized AgNPs and induce their aggregation. The hydrolysis catalyzed with trypsin in sample solution decreased the concentration of free protamine, resulting in the dispersion of AgNPs and thus decreasing the Raman intensity of 4-MPY, by which the trypsin could be sensed optically. A detection level down to 0.1 ng mL−1 for trypsin was obtained. The induced accumulation of AgNPs modified with Raman reporter 4-MPY largely enhanced the SERS responses. A good linearity was found within the wide range over five orders of magnitude and reasonable relative standard deviations (between 2.4 and 11.6%) were attained. By using trypsin as a model, the new concept can provide an excellent platform for ultrasensitive SERS measurements of various proteases/enzymes which can lead to nanoparticles stability change through catalyzed hydrolysis toward substrate.
研究领域:
生物化学
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分析化学
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纳米技术
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DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
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2023
·
IF 15.1
Jin-Zheng Wang,Cheng-Kun Wu,Xin-Ying Wang,Yuan-Zhi Xu,Richard Ansah Herman,Sheng Sheng,Shuai You,Jun Wang,Fu-An Wu
Edible insects are rich in oil and protein resources, and has it become increasingly important to develop them reasonably when resources are increasingly scarce. However, due to the challenges in recovering enzymes in the enzyme-assisted aqueous extraction technique and the remaining organic solvents in the extraction process based on organic solvent, it is difficult to achieve simultaneous utilization of oil and protein from edible insects. Hence, an improved enzyme-assisted aqueous extraction process based on the assistance of chitosan cross-linked Fe 3 O 4 nanoparticles (CNPs) immobilized enzymes and divalent copper ions chelation, [email protected] aqueous extraction technique, was first proposed. The hydrolysis activity was significantly increased by 6.16 times, and the value of K cat / K m was increased by 1.7 times for the results of the formation of the hydrophilic microenvironment. Furthermore, the extraction ratio of silkworm pupae oil extracted via [email protected] aqueous extraction technique could be increased to 93%, and the oil extraction rate could be maintained above 65% after 10 times reused. Improvement of enzyme-assisted aqueous extraction under the addition of divalent copper ions could achieve an ideal effect of oil extracted from silkworm pupae with less time. Additionally, the quality of silkworm pupae oil was significantly improved and the silkworm pupae protein hydrolysate also showed improved biological activity. Therefore, the enzyme-assisted aqueous extraction enhancement by trypsin immobilization and divalent copper ion chelation is a promising strategy for the green and sustainable exploration of edible insect oil and protein without the use of organic solvents.
研究领域:
食品科学
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生物化学
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纳米技术
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酶学
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可持续资源利用
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DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2024
·
IF 8
Yiyue Tang,Xiaomei Mo,Lili Yuan,Xueer Huang,Yongkang Yan,Wenying Jin,Lang Zhang,Xinqing Jiang,Jinfang Nie,Yun Zhang
The classical colorimetric assays with Au nanoparticles (AuNPs) are not suitable for analytical systems where the analyte could not induce a red-to-blue color change related to their surface plasmon resonance (SPR). To respond this challenge, herein this study utilizes Tyndall effect (TE) of AuNPs as amplified colorimetric signaling to design a universal visual assay for sensitive detection of trypsin (model analyte). The AuNPs are modified with biotinylated peptides. In the absence of trypsin, streptavidin binds biotin on these probes to form a network-like complex consisting of repeated units of “AuNP-biotin-streptavidin-biotin-AuNP” resulting in a mixture that shows an intense TE response as well a SPR-related red color. Upon trypsin incubation, however, the analyte can cleave the peptide chain to inhibit the streptavidin-mediated production of network-like complex; the resultant red reaction solution containing free AuNP probes would display a weak TE signal. The reduction degree in the TE positively relies on the trypsin level in sample. The results demonstrate that this new equipment-free method only requires a laser pointer pen (for the TE creation) and a smartphone (for portable TE imaging readout) to linearly detect trypsin in a concentration range of 3.9–1000 nM, with an estimated limit of ∼0.63 nM (3σ). The satisfactory recovery of trypsin in actual human serum samples is between 94.93–102.21 %. Such TE-enhanced strategy may significantly expand theoretical scope of widely used AuNP-based visual assays and their applications in various resources-limited settings like point-of-care testing of trypsin for medical diagnosis.
研究领域:
分析化学
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生物医学
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诊断学
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高影响因子
JOURNAL OF SEPARATION SCIENCE
·
2019
·Vol. 42
·期 11
·1980-1989
·
IF 2.516
Hang Lin,Changfa Zhang,Yuanjing Lin,Yiqun Chang,Jacques Crommen,Qiqin Wang,Zhengjin Jiang,Jialiang Guo
A novel strategy was successfully developed for screening trypsin inhibitors in traditional Chinese medicines based on monolithic capillary immobilized enzyme reactors combined with liquid chromatography-tandem mass spectrometry. Organic polymer based monolithic enzyme reactors were firstly prepared by covalently bonding trypsin to a poly(glycidyl methacrylate- co -poly (ethylene glycol) diacrylate) monolith by the ring-opening reaction of epoxy groups. The activity and kinetic parameters of the obtained monolithic trypsin reactors were systematically evaluated using micro-liquid chromatography. Fourier transform infrared spectroscopy and scanning electron microscopy were also used to characterize the monolithic trypsin reactors. The resulting functional and denatured monolithic trypsin reactors were applied as affinity solid-phase extraction columns, and offline coupled with a liquid chromatography-tandem mass spectrometry system to construct a binding affinity screening platform. Subsequently, the proposed platform was applied for screening trypsin binders in a Scutellaria baicalensis Georgi extract. Three compounds, namely scutellarin, baicalin, and wogonoside were identified, and their inhibitory activities were further confirmed via an in vitro enzymatic inhibition assay. Additionally, molecular docking was also performed to study the interactions between trypsin and these three compounds.
研究领域:
药物分析
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天然产物化学
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酶工程
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分析化学
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中药研究
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DOI
高影响因子
CHEMOSPHERE
·
2023
·
IF 8.8
Qinyuan Lu,Siyuan Wang,Qian Ping,Yongmei Li
A novel approach with a combination of trypsin, nano-zero-valent iron (NZVI) and activated carbon (AC) was conducted to promote the methane production of waste activated sludge (WAS) during the anaerobic digestion (AD) processes. Results showed that the combined addition of trypsin-NZVI-AC exhibited the synergistic effect during different AD stages. Trypsin mainly facilitated the hydrolysis process and the acetic acid conversion, while NZVI-AC enhanced the substrate metabolism and the electronic transfer to subsequently produce methane. A dose of 1000 mg/L trypsin was optimal to maximize this synergistic effect. Metagenomic analysis showed that trypsin-NZVI-AC addition effectively improved the relative abundance of acetyl-CoA carboxylase, and then strengthened both acetoclastic methanogenesis (M00357) and hydrogenotrophic methanogenesis (M00567). Hydrogenotrophic methanogens such as Methanobacterium, Methanoculleus, and Methanosarcina were greatly enriched with trypsin-NZVI-AC compared with trypsin or NZVI-AC addition. Moreover, electroactive bacteria G. sulfurreducens and G. metallireducens were also enriched by this method to conduct direct interspecies electron transfer among methanogens , leading to the better improvement of methane production. These findings supply a promising way to optimize the enzyme pretreatment technology and elevate the methanogenic efficiency of WAS.
研究领域:
环境科学
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微生物学
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DOI
PubMed
高影响因子
Food Chemistry-X
·
2026
·
IF 9.2
Hongyu Chen,Yafang Wang,Xinrui Sun,Nannan Wang,Wenping Zhu,Manman Sun,Qingfeng Li,Yahong Chen
Combining the excellent optical properties, spatial structure specificity and 30-fold cascade signal amplification strategy of the trypsin-stabilized silver‑gold nanoflowers (Try-AgAuNFs), a novel nanosensing system was constructed for improving the sensing of hydrogen sulfide (H 2 S). It is worth noting that the high loading capacity of Ag can be used both for the design of the cascade signal amplification strategy, and to effectively increase the active sites of the probe, making it suitable for real-time detection of H 2 S. Excitingly, the Try on the surface of Try-AgAuNFs increases the spatial steric hindrance of the probe, effectively preventing biothiols and other amino acids from interacting with Try-AgAuNFs, thereby enabling reliable H 2 S detection and food-freshness evaluation across diverse samples. This work not only presents a novel and highly stable method for synthesising Try-AgAuNFs, but also reveals that surface-bound protein ligands can enhance probe selectivity through steric hindrance control.
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PubMed
高影响因子
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