ACS Applied Materials & Interfaces
·
2020
·
IF 8.758
Jiaojiao Su,Haixu Chen,Zhijun Xu,Shengliang Wang,Xiaoman Liu,Lei Wang,Xin Huang
Inspired by the compartmentalized structure of cells, self-regulating responsive hollow microcapsules are highly desirable for the modulation of enzymatic reactions. Here, we report a strategy to fabricate gold nanorod embedded proteinosomes by covalently grafting gold nanorods onto the surface of proteinosomes. The excellent photothermal conversion efficiency of the embedded gold nanorod and the thermal phase transition of the grafted PNIPAAm allow the constructed hybrid proteinosomes to show reversible contraction behaviors triggered by near-infrared light with the molecular weight cutoff of the membrane decreased to ca. 50 kDa, and importantly, the contraction frequency of the constructed proteinosomes could be as fast as 1 min and last for at least 15 cycles. Subsequently, the effective encapsulation of three cascade enzymes into the proteinosomes realizes the construction of a near-infrared responsive microreactor that allows control of the cascade reaction by near-infrared illumination, thereby enabling reversible on and off of the enzymatic reaction. Such microcapsule-based reactors demonstrate the potential to alter the membrane molecular weight cutoff, and it is believed that the development of such responsive microcapsules will have great potential for studying cellular responses and provide a platform for future applications in biosensing and drug delivery.
研究领域:
生物工程
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材料科学
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生物医学工程
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DOI
PubMed
高影响因子
Current Pharmaceutical Analysis
·
2023
·Vol. 19
·期 3
·216-221
·
IF 0.6
Zhang Liang,Yang Dehui,Liu Ye
Background: A reversed-phase high-performance liquid chromatographic method for determining thrombin activity is described. The improved method can provide a high resolution between substrate Nα-P-toluenesulfonyl-L-arginine methyl ester (TAME) and the product of enzyme reaction,
Nα-P-toluenesulfonyl-L-Arginine (TA). Methods: When the substrate concentration was 5 mg/mL, the Michaelis equation approached the zero-order reaction, and the thrombin enzyme activity must have a good linear relationship with the consumption of TAME or the yield of TA. Results:
The method is suitable for quantitatively analyzing thrombin activity in formulation or serum. Conclusion: Three batches of thrombin lyophilized preparations were determined for activity.
研究领域:
生物化学
·
医学检测
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DOI
高影响因子
APPLIED OPTICS
·
2019
·Vol. 58
·期 17
·4806-4811
·
IF 1.973
Jianyang Hu,Dongying Fu,Chunli Xia,Sibang Long,Chunlian Lu,Weimin Sun,Yongjun Liu
A novel, to the best of our knowledge, liquid crystal (LC) biosensor, based on an optical fiber Mach–Zehnder interferometer (MZI), is proposed. The proposed optical fiber MZI consists of two single-mode fibers and a tapered photonic crystal fiber (PCF). The PCF is coated with 4′-pentyl-biphenyl-4-carboxylic acid (PBA)-doped 4-cyano-4′-pentylbiphenyl (5CB). Being a pH-sensitive material, PBA can manipulate LC molecules to different orientations according to their pH values. When the orientation of LC molecules changes with varying pH, the effective refractive index of the cladding modes also is accordingly affected. Enzymatic reactions of penicillinase can release H+, which causes the decrease of the pH. Therefore, the enzymatic reactions of penicillinase can be sensed by monitoring the peak shift in the interference spectrum. The effects of the tapered diameter on the sensitivity of the sensor were experimentally investigated as well.
研究领域:
生物传感
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光学工程
·
生物化学
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PubMed
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
·
2019
·
IF 3.371
Yang Hu,Yan Sun
Autonomous motion propelled by the reactions of enzymes asymmetrically immobilized on Janus particles have attracted great attention due to its possible auspicious application as active biocompatible platforms. However, no research has focused on the effect of the autonomous motion on enzyme reactions. Hence, this work was designed to fabricate different enzyme-immobilized particles with asymmetrical (Janus) and symmetrical enzyme distributions and to investigate the interplay between enzymatic reactions and their motion behavior with lipase and urease as model enzymes. It was found that all the EIPs displayed substrate-concentration-dependent enhanced diffusions, but Janus enzyme-immobilized particles exhibited maximum enhancement (60%) of autonomous motion represented by diffusion coefficient as well as superior specific activity increments (>2 times). Namely, the enzymatic reactions were more significantly facilitated by the autonomous motions of the Janus enzyme-immobilized particles. The results indicated that the asymmetrical enzyme distribution on Janus particles enhanced the favorable mutual promotion between the self-propelled movement and enzymatic reactions, leading to improved diffusional and enzymatic performance. The results have thus provided insights into the design of new materials for enzyme immobilization and novel immobilized enzyme systems for various applications.
研究领域:
生物化学
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材料科学
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DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2017
·
IF 5.401
Yan Wang,Liyuan Zhao,Aojie Xu,Lei Wang,Lingli Zhang,Shuangqiang Liu,Yongjun Liu,Hanyang Li
We report a liquid crystal (LC) microdroplet-based pH sensor and explore its application for detecting enzymatic reactions in penicillinase. Nematic LC 4-cyano-4'-pentylbiphenyl (5CB) microdroplet, doped with 4'-pentyl-biphenyl-4-carboxylic acid (PBA), is produced in phosphate buffer solution (PBS) with a tapered capillary microtube connected to a syringe pump. The PBA-doped 5CB microdroplets exhibit configuration transitions from bipolar to radial as PBS pH values increase from 5.7 to 6.0. These pH-induced changes in optical appearance are experimentally verified and attributed to the deprotonation of PBA at the aqueous/LC interface. We also examine the feasibility of the PBA-doped 5CB microdroplets as optical microcavities of whispering gallery modes (WGMs). Typical WGM lasing emission is observed in micrometer-sized microdroplets under a pulse laser pump. The WGM spectrum from the microdroplet responds to pH variations, reflecting as a spectral redshift as the pH decreases. The PBA-doped 5CB microdroplet can be successfully exploited accordingly for detecting enzymatic reactions in penicillinase. We demonstrate that the detectable shifts in WGM spectra associated with the microdroplet configuration transitions can serve as an indicator of the H + released from the enzymatic reaction.
研究领域:
生物化学
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分析化学
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材料科学
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DOI
高影响因子
TALANTA
·
2019
·
IF 4.916
Rui Duan,Yanzeng Li,Bojian Shi,Hanyang Li,Jun Yang
In this study, we demonstrate a novel nematic liquid crystal (LC) 4-cyano-4′-pentylbiphenyl (5CB) microdroplet-based biosensor for real-time, quantitative and sensitive detection of urea by whispering gallery mode (WGM) lasing technology. The single stearic acid-doped 5CB microdroplet is taken as both optical resonator and sensing reactor. To the best of our knowledge, this work is the first report of urea detection with WGM lasing. The enzymatic reaction between urea and urease produces hydroxide ions. Deprotonation and self-assembly of stearic acid occur at the aqueous/LC interface with increased pH value, promoting the reorientation of LC molecules. The detectable shift of WGM lasing spectra related to the configuration transition of LC microdroplets can be used as an indicator of enzymatic reaction, and this allows detection of urea molecules by the sensor at a concentration of only 0.1 mM. The proposed sensor also demonstrated the ability to detect urea in real urine samples. Compared with conventional POM observations, monitoring and identification of lasing spectra from LC microdroplets as a versatile method allow for more quantitative and sensitive detection of analyte reactions and can be expected to replace POM imaging technology for current LC-based biosensor systems.
研究领域:
生物传感
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光学检测
·
分析化学
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DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY A
·
2015
·
IF 4.169
Lulu Shangguan,Lingyi Zhang,Zhichao Xiong,Jun Ren,Runsheng Zhang,Fangyuan Gao,Weibing Zhang
The bottom-up strategy of proteomic profiling study based on mass spectrometer (MS) has drawn high attention. However, conventional solution-based digestion could not satisfy the demands of highly efficient and complete high throughput proteolysis of complex samples. We proposed a novel bi-enzymatic reactor by immobilizing two different enzymes (trypsin/chymotrypsin) onto a mixed support of hybrid organic–inorganic monolith with SBA-15 nanoparticles embedded. Typsin and chymotrypsin were crossly immobilized onto the mixed support by covalent bonding onto the monolith with glutaraldehyde as bridge reagent and chelation via copper ion onto the nanoparticles, respectively. Compared with single enzymatic reactors, the bi-enzymatic reactor improved the overall functional analysis of membrane proteins of rat liver by doubling the number of identified peptides (from 1184/1010 with trypsin/chymotrypsin enzymatic reactors to 2891 with bi-enzymatic reactor), which led to more proteins identified with deep coverage (from 452/336 to 620); the efficiency of the bi-enzymatic reactor is also better than that of solution-based tandem digestion, greatly shorting the digestion time from 24 h to 50 s. Moreover, more transmembrane proteins were identified by bi-enzymatic reactor (106) compared with solution-based tandem digestion (95) with the same two enzymes and enzymatic reactors with single enzyme immobilized (75 with trypsin and 66 with chymotrypsin). The proteolytic characteristics of the bi-enzymatic reactors were evaluated by applying them to digestion of rat liver proteins. The reactors showed good digestion capability for proteins with different hydrophobicity and molecular weight.
研究领域:
生物化学
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蛋白质组学
·
分析化学
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DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY A
·
2016
·
IF 3.926
Bingbing Wang,Lulu Shangguan,Shulei Wang,Lingyi Zhang,Weibing Zhang,Fan Liu
The bottom up strategy has drawn much attention due to the high accuracy, reliability, and reproducibility in protein identification in which proteins are digested into peptides. However, conventional solution-based digestion and enzymatic reactor with one protease immobilized cannot satisfy high throughput proteolysis of complex samples. Application of consecutive hydrolysis by enzymatic reactor can be a new strategy for high throughput proteolysis of complex samples by adjusting immobilization amount of the enzymes, enzyme ratio, as well as hydrolysis order of two enzymes. In this work, we propose immobilized enzymatic reactor for consecutive digestion with two enzymes by combining two enzyme reactors with trypsin and chymotrypsin immobilized, respectively. Each reactor was prepared individually by immobilizing only one protease (trypsin or chymotrypsin) to hybrid monolith with SBA-15 particles embedded. Proteolysis conditions including hydrolysis order and trypsin to chymotrypsin ratio etc. were studied using standard proteins. Best digestion performance was obtained when the proteins were digested by trypsin first with trypsin to chymotrypsin ratio of 1:1. When applying them to digestion of rat liver proteins, total 1651 proteins and 11011 peptides were identified by combining four enzymolysis strategies with two enzymes including proteolytic digestion in two consecutive enzymatic reactors, synergy enzymolysis with two enzymes in one immobilized enzymatic reactor and consecutive hydrolysis with two enzymes in-solution digestion respectively, in which consecutive enzymolysis in enzymatic reactors gave the best results with 1091 proteins and 5071 peptides identified. The reactors showed good digestion capability for proteins with different hydrophobicity and molecular weights, and will play an important role in high efficient and high throughput proteomics research.
研究领域:
生物化学
·
蛋白质组学
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DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2019
·
IF 8.456
Juan Qiao,Junfang Jiang,Lili Liu,Ji Shen,Li Qi
The challenge for polymeric enzyme reactors at present is to selectively control the enzymolysis rate in complex conditions. Additionally, the fabrication methodology is hindered by complex processes, especially for achieving diverse stimuli responsiveness and functions. Here, we reported a kind of pH-sensitive polymer, poly(styrene-co-maleic anhydride-acrylic acid) (PS-MAn-AA)-based hybrid enzyme reactor. It comprised magnetic nanoparticles and a pH-sensitive PS-MAn-AA porous polymer membrane made by breath figure method. The enzyme l-asparaginase (l-ASNase) could covalently bond on the surface of the pH-sensitive porous polymer membrane (pH-PPM), and the resultant enzyme reactor was characterized by Fourier transform infrared spectroscopy and vibrating sample magnetometer. The apparent Michaelis–Menten constants (Km and Vmax) of the l-ASNase enzyme reactor at different pH values were determined by a chiral ligand-exchange capillary electrophoresis method with l-asparagine as the substrate. The Vmax value of the l-ASNase enzyme reactor (0.67 mM/min) was almost 3-fold of that of the free l-ASNase (0.23 mM/min) at pH 8.2. Its ability to precisely control the enzymolysis rate in complex conditions is triggered primarily by the pH of the buffer solution, allowing controlled enzymatic reactions and displaying excellent stability and reusability of the proposed pH-PPM. This strategy for porous polymer membrane enzyme reactor fabrication has established a platform for enzyme efficiency adjusting. These valve-like distinguished features highlight the outstanding potential of stimuli-responsive enzyme reactor applied for enzyme immobilization and enzyme-related disease treatment.
研究领域:
生物化学
·
材料科学
·
酶工程
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DOI
PubMed
高影响因子
JOURNAL OF SEPARATION SCIENCE
·
2014
·Vol. 37
·期 22
·3377-3383
·
IF 2.594
Tingting Hong,Cuijie Chi,Yibing Ji
Pepsin-modified affinity monolithic capillary electrochromatography, a novel microanalysis system, was developed by the covalent bonding of pepsin on silica monolith. The column was successfully applied in the chiral separation of (±)-nefopam. Furthermore, the electrochromatographic performance of the pepsin-functionalized monolith for enantiomeric analysis was evaluated in terms of protein content, pH of running buffer, sample volume, buffer concentration, applied voltage, and capillary temperature. The relative standard deviation (%RSD) values of retention time (intraday <0.53, n = 10; interday <0.53, n = 10; column-to-column <0.70, n = 20; and batch-to-batch <0.80, n = 20) indicated satisfactory stability of these columns. No appreciable change was observed in retention and resolution for chiral recognition of (±)-nefopam in 50 days with 100 injections. The proteolytic activity of this stationary phase was further characterized with bovine serum albumin as substrate for online protein digestion. As for monolithic immobilized enzyme reactor, successive protein injections confirmed both the operational stability and ability to reuse the bioreactor for at least 20 digestions. It implied that the affinity monolith used in this research opens a new path of exploring particularly versatile class of enzymes to develop enzyme-modified affinity capillary monolith for enantioseparation.
研究领域:
分析化学
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生物化学
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DOI
PubMed
高影响因子
Food & Function
·
2020
·Vol. 11
·期 5
·4356-4362
·
IF 4.171
Yunci Zhang,Dongyu Gu,Shuai He,Jing Meng,Jifeng Wang,Yunxiao Wang,Yi Wang,Jing Tian,Yi Yang
The flower of S. japonica is a favorite food and used as traditional medicine. In the present study, a facile and effective method based on the changes in the composition before and after the enzyme reaction was established to screen the active compounds from complex natural products. The separation of an active compound from the ethanolic extracts of Sophora japonica var. violacea, which exhibited the α-amylase inhibitory activity is presented as an example. The analysis of HPLC showed that one component was reduced by 25% after the enzyme reaction. The potential active compound was isolated via LH-20 gel permeation chromatography and identified as kaempferol 3-O-α-L-rhamnopyranosyl-(1 → 6)-β-D-galactopyranosyl-7-O-α-L-rhamnopyranoside by 1H and 13C NMR. The in vitro test indicated that the compound had the α-amylase inhibitory activity, and the IC50 was 88.56 ± 0.60 μg mL−1. The molecular docking study of this compound showed that the compound enfolded in the active sites of α-amylase completely and interacted with the amino acid residues through hydrogen bonds, van der Waals force and hydrophobic interactions.
研究领域:
药理学
·
植物化学
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DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2025
·
IF 9.8
Shuo Liu,Hongli Chao,Meishan Guo,Ruifang Wang,Dongyu Gu,Yi Wang,Dajun He,Yi Yang
To achieve the biotransformation of cellulose into gluconic acid, a multi-enzyme reactor comprising cellulase, glucose oxidase, and catalase was constructed through artificial antibody-antigen-directed immobilization. Catechol was used as a template to prepare artificial antibodies, which were then used to modify the enzymes through a Schiff base reaction to form artificial antigens. The artificial antibody and antigen were subsequently assembled via affinity recognition to immobilize the enzymes. The immobilization yields for all three enzymes exceeded 85 %, with specific activities of 43.48 ± 1.54 U/mg, 0.88 ± 0.06 U/mg, and 18.47 ± 1.43 U/mg, respectively. The immobilized enzymes demonstrated excellent adaptability, storage stability, reusability, and regenerability. Through the modular assembly of artificial antigen-antibody, the multi-enzyme reactor facilitated the cascade conversion of cellulose to gluconic acid.
研究领域:
生物化学
·
合成生物学
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产品关联
DOI
PubMed
高影响因子
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