ANALYTICA CHIMICA ACTA
·
2025
·
IF 6
Juan Li,Ming-Chun Lai,Ya-Meng Zhong,Ya-Ling Chen,Na Wu,Wei Chen,Hua-Ping Peng
Monitoring D-amino acids concentrations has essential implications for gastric cancer diagnosis and treatment, especially in the non-invasive detection of gastric cancer. However, it remains a challenge to establish a high-performance detection method for D-amino acids. Here, a highly sensitive and selective d -alanine (D-Ala) ECL assay strategy via a turn-off-on split-type electrochemiluminescence (ECL) platform has been proposed. We found that hydrogen peroxide (H 2 O 2 ) could be used as an efficient etching agent to turn on the MnO 2 /gold nanocluster (AuNC)-based ECL nanoswitch platform. Based on abovementioned characteristics, we extended it to the D-amino acids assay since the enzymatic reaction between D-Ala and D-amino acid oxidase (DAAO) generates H 2 O 2 . Based on the abovementioned characteristics, this ECL sensing platform achieved a preferable linear-dependent curve in the detection range of 1.0 × 10 − 10 ∼1.0 × 10 − 3 mol L −1 , and realized the detection of D-Ala as low as 2.2 × 10 − 11 mol L −1 (S/N = 3). Furthermore, the proposed ECL biosensor showed excellent selectivity, stability, and reproducibility. Together with its powerful performance, this strategy could test D-Ala in saliva samples, which suggested that this ECL assay platform shows great prospect in disease diagnosis. We ascribe the high sensitivity and good anti-interference capability of the sensor to the combination of specific enzyme catalysis reaction, high-efficiency split-type AuNC probe-based ECL technique and the highly etching efficiency of H 2 O 2 to MnO 2 nanomaterials on electrode surface. In our perception, this H 2 O 2 mediated split-type ECL sensing platform provides a viable tool in ECL based bioananlysis. Therefore, our proposed approach not only provides a strategy for developing a high-performance platform for D-Ala detection, but also establishes a framework for the detailed design and development of ECL platform for other biological assays.
研究领域:
生物医学
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分析化学
·
癌症诊断
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产品关联
DOI
PubMed
高影响因子
Theranostics
·
2018
·
IF 8.537
Xinglong Yang,Yan Dang,Jinli Lou,Huawu Shao,Xingyu Jiang
Rationale: Rapid and facile detection of pathogenic bacteria is challenging due to the requirement of large-scale instruments and equipment in conventional methods. We utilize D-amino acid as molecules to selectively target bacteria because bacteria can incorporate DADA in its cell wall while mammalian cells or fungi cannot. Methods: We show a broad-spectrum bacterial detection system based on D-amino acid-capped gold nanoparticles (AuNPs). AuNPs serve as the signal output that we can monitor without relying on any complex instruments. Results: In the presence of bacteria, the AuNPs aggregate and the color of AuNPs changes from red to blue. This convenient color change can distinguish between Staphylococcus aureus ( S. aureus ) and methicillin-resistant Staphylococcus aureus (MRSA). This system can be applied for detection of ascites samples from patients. Conclusion: These D-amino acid-modified AuNPs serve as a promising platform for rapid visual identification of pathogens in the clinic.
研究领域:
生物医学工程
·
微生物学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2024
·
IF 6.7
Chang Liu,Yanan Wu,Mengying Li,Fang Liu,Pengfei Kong,Haifeng Yang,Xinling Liu
Salivary d-alanine (d-Ala) and d-proline (d-Pro) are of concern for their potential in the noninvasive diagnosis of gastric cancer (GC). Most reports have succeeded in determining the total concentration of d-Ala and d-Pro. However, for personalized diagnosis and better elucidation of the underlying specific correlation of d-Ala (or d-Pro) with GC, it is desirable to determine the specific concentration of d-Ala or d-Pro. Herein, we propose an enantiomer-specific tandem assay of d-Ala based on the colorimetric reaction between 2,4-dinitrophenylhydrazine and pyruvic acid generated from the deamination of d-Ala catalyzed by d-amino acid oxidase, which is easily distinguished from l-form amino acids, d-Pro, and many other species. A linear concentration range is established from 20 to 400 μmol/L with a limit of detection of 1.01 μmol/L. Real saliva sample tests reveal that the levels of d-Ala in GC cases are remarkably higher than those in healthy individuals, which offers a simple and low-cost strategy for GC diagnosis. Simultaneously, the total concentrations of d-Ala and d-Pro in saliva are determined. Hence, the concentration of d-Pro and the proportion of d-Ala could be calculated, which further provides more molecule- and individual-specific information. This research may offer a convenient method for noninvasive diagnosis of GC and pave a new route to explore the potentials of rare d-form amino acids in disease diagnosis and treatment.
研究领域:
生物化学
·
医学诊断
·
癌症研究
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2019
·
IF 6.393
Han Liu,Juanjuan Shao,Lixia Shi,Wei Ke,Fangjie Zheng,Yuan Zhao
A simple and accurate magnetic electrochemical sensor is developed for the ultrasensitive chiral recognition of D-amino acids (D-AAs). Fe 3 O 4 @Au@Ag@Cu x O nanoparticles (NPs) (x = 1, 2) are controllable prepared and used as the electrochemical beacons. The controllable integration of plasmonic metal NPs, semiconductors and magnetic NPs enables multilayered Fe 3 O 4 @Au@Ag@Cu x O NPs to display a sharp copper stripping peak at -0.16 V, owning to the electron transfer from Cu + to Cu 2+ and the synergistic electron transfer of Au and Ag double layers. D-AAs can be catalyzed by D-amino acid oxidase (DAAO) to produce H 2 O 2 . The autocatalytic oxidation reaction of Cu x O shell with the existence of H 2 O 2 induces the decrease of electrochemical signals of Fe 3 O 4 @Au@Ag@Cu x O NPs. Fe 3 O 4 @Au@Ag@Cu x O NPs served as electrochemical beacons achieve the sensitive and accurate enantioselective recognition of D-alanine (D-Ala) in the range from 100 pM to 10 μM. The limit of detection (LOD) is as low as 52 pM. This developed protocol can be extended to the fabrication of a large set of electroactive labels by the introduction of plasmonic metal NPs with high electron transfer efficiency, for the reliable enantioselective recognition with high sensitivity.
研究领域:
生物化学
·
分析化学
·
材料科学
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产品关联
DOI
高影响因子
NEW JOURNAL OF CHEMISTRY
·
2021
·Vol. 45
·期 43
·20419-20425
·
IF 3.591
Siqi Chen,Tingting Li,Donglian Deng,Xiaoyue Zhang,Yibing Ji,Ruijun Li
The integration of paper-based devices with fluorescent nanomaterials as recognition elements has offered novel opportunities for biosensing with enhanced sensitivity and selectivity, as well as stable signals. This work innovatively designed a fluorescent paper-based device for enantioselective sensing of D-alanine (D-Ala), where carbon dots (CDs) grafted cellulose paper (PCD) was used as the fluorophore and the in situ generated gold nanoparticles (AuNPs) were used as quenchers. D-α-Amino acid oxidase (DAAO) stereo-selectively catalyzed D-Ala to produce hydrogen peroxide (H2O2), which mediated the in situ formation of PCD/AuNPs composites and quenched the fluorescence of PCD based on fluorescence resonance energy transfer (FRET). Under optimal conditions, the designed PCD/AuNPs demonstrated a reliable quenching response to D-Ala in the range of 1–100 μM and the limit of detection was 0.55 μM. Moreover, the platform was applied to detect D-Ala in serum, simulated gastric fluid samples, and was further employed to determine D-Ala in BGC-823 gastric cancer cells with satisfactory results. Ultimately, the PCD/AuNPs platform provided a novel strategy for the development of reliable and sensitive assays based on paper-based devices. It has great potential to be utilized in enantioselective sensing and clinical early diagnosis.
研究领域:
分析化学
·
生物医学
·
材料科学
✓ 已验证
产品关联
DOI
高影响因子
Materials Science & Engineering C-Materials for Biological Applications
·
2019
·
IF 4.959
Yumei Wang,Wei Li,Tiaotiao Liu,Lu Xu,Yingyu Guo,Jia Ke,Sanming Li,Heran Li
In this study, twisted rod-like chiral mesoporous silicas (CMSs) with discriminating chiral characteristics (D/L) were designed and biomimetic synthesized by using L- and d -alanine derivatives as templates, and employed as poorly water-soluble chiral drug ibuprofen (IBU) carriers. The morphology and mesoscopic characteristics of CMSs were determined by transmission electron microscope (TEM) and small-angle X-ray scattering (SAXS). Meanwhile, the physicochemical properties of CMSs before and after drug loading were systematically characterized by infrared spectroscopy (IR), nitrogen adsorption, X-ray diffraction (XRD), differential scanning calorimetry (DSC) and induced circular dichroism (ICD). The results suggested that, the CMSs exhibited local chiral characteristics, which were successfully endowed by the alanine-derivative surfactants templates with a reversal of chirality. The crystalline state of IBU was effectively converted to amorphous state after being incorporated into CMSs, and the drug delivery systems shared the chiral characteristic of carriers. Besides, in vitro drug release experiments were respectively performed in simulated gastric fluid (SGF, pH 1) and simulated intestinal fluid (SIF, pH 6.8) medium, and the results demonstrated that both l -CMS and d -CMS could improve the dissolution of IBU in SGF medium, which could be explained by the amorphization of IBU. Particularly, due to the different pore geometry of these two materials, CMSs with different chirality (D/L) could be applied as carriers to accomplish drug release differentiation.
研究领域:
材料科学
·
药学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF THE ELECTROCHEMICAL SOCIETY
·
2016
·Vol. 163
·期 7
·B373
·
IF 3.014
Qinghong Wang,Xia Lin,Juanjuan Xu,Chunzhi Xuan,Shu Zhu,Yingzi Fu
A new biosensor specified for D-amino acids (DAAs) based on the electrochemiluminescence (ECL) of luminol was proposed by coupling D-amino acid oxidase (DAAO) to in situ generate coreactant with functionalized Multi-walled carbon nanotubes (F-MWCNTs) and Au-Pt alloy nanoparticles as catalysts for the ECL reaction. The stepwise fabrication process of the biosensor was characterized via cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and scanning electron microscopy (SEM). Furthermore, the Au–Pt nanostructures with different molar ratios between Au and Pt precursors were investigated by scanning electron microscopy (SEM), energy-dispersive spectroscopy (EDS) and ECL technique, and the optimum Au/Pt ratio for ECL measurements was 1:1 with an optimized electrodeposition time at 400 s. The proposed biosensor showed excellent performance in the stereoselective recognition of D-alanine with a low detection limit of 1.67 × 10−9 M. Hence, this paper would provide a new platform on the specific detection of D-alanine with high sensitivity, good selectivity and acceptable stability.
研究领域:
生物化学
·
分析化学
·
纳米技术
✓ 已验证
DOI
高影响因子
Analytical Methods
·
2017
·Vol. 10
·期 1
·84-90
·
IF 1.9
Xia Lin,Shu Zhu,Qiao Xia,Jiao Ma,Yingzi Fu
Herein, an ultrasensitive and stereo-selective electrochemiluminescent (ECL) biosensor based on ECL signal amplification of luminol by the synergetic catalysis of a hemin-functionalized composite and gold–platinum nanowires (Au-PtNWs) has been designed for the detection of D-alanine (D-Ala). In the sensor design, D-amino acid oxidase (DAAO) was used to generate the ECL co-reactant (H2O2) for luminol in situ. The reduced graphene oxide-hemin-cysteine (rGO-H-Cys) composite and Au-PtNWs acted not only as amplification labels to amplify signals via synergetic catalysis, but also as an ideal nanocarrier to accelerate electron transfer. A great difference in ECL intensities toward D-Ala and L-Ala was obtained, and an obviously higher intensity was seen for D-Ala. Under optimal conditions, the ECL intensity had a linear relationship with the logarithm of the D-Ala concentration in the range from 1.0 × 10−9 to 5.0 × 10−3 M with a detection limit of 3.3 × 10−10 M (S/N = 3). This study provides a new method for the determination of D-Ala in food with high sensitivity, excellent selectivity, and good stability and reproducibility.
研究领域:
分析化学
·
生物医学工程
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产品关联
DOI
高影响因子
ENGINEERING IN LIFE SCIENCES
·
2019
·Vol. 19
·期 4
·270-278
·
IF 1.936
Jinyuan Hu,Xiao Lu,Haokun Wang,Fuxiang Wang,Yuan Zhao,Wei Shen,Haiquan Yang,Xianzhong Chen
d -Alanyl- d -alanine carboxypeptidase DacC is important for synthesis and stabilization of the peptidoglycan layer of Escherichia coli . In this work, dacC of E . coli BL21 (DE3) was successfully deleted, and the effects of this deletion on extracellular protein production in E . coli were investigated. The extracellular activities and fluorescence value of recombinant amylase, green fluorescent protein, and α-galactosidase of the deletion mutants were increased by 82.3, 29.1, and 37.7%, respectively, compared with that of control cells. The outer membrane permeability and intracellular soluble peptidoglycan accumulation of deletion mutant were also enhanced compared with those of control cells, respectively. Based on fluorescence-assisted cell sorting analyses, we found that the morphology of the E. coli deletion mutant cells was altered compared with that of control cells. Local transparent bulges in the poles of the E . coli mutant with deletion of the dacC gene were found by transmission electron microscopy analysis. These bulges in the poles could explain the improvement in the production of extracellular protein by the E. coli mutant with deletion of the dacC gene. These findings provide important insights into the extracellular production of proteins using E . coli as microbial cell factories.
研究领域:
微生物学
·
生物技术
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
ORGANIC PROCESS RESEARCH & DEVELOPMENT
·
2022
·
IF 3.858
Jianjiong Li,Shanshan Yu,Yingang Wang,Peiyuan Yao,Qiaqing Wu,Dunming Zhu
(S)-2-Aminobutane, d-alanine, and d-homoalanine are important intermediates for the production of various active pharmaceutical ingredients and food additives. The preparation of these small chiral amine or amino acids with high water solubility still demands searching for efficient methods. In this work, we identified an ω-transaminase (ω-TA) from Sinirhodobacter hungdaonensis (ShdTA) that catalyzed the kinetic resolution of racemic 2-aminobutane at a concentration of 800 mM using pyruvate as the amino acceptor, leading to the simultaneous isolation of enantiopure (S)-2-aminobutane and d-alanine in 46% and 90% yield, respectively. In addition, (S)-2-aminobutane (98% ee) and d-homoalanine (99% ee) were isolated in 45% and 93% yield, respectively, in the kinetic resolution of racemic 2-aminobutane at a concentration of 400 mM coupled with deamination of l-threonine by threonine deaminase. We thus developed a biocatalytic process for the practical synthesis of these valuable small chiral amine and d-amino acids.
研究领域:
生物催化
·
有机化学
·
药物化学
✓ 已验证
DOI
高影响因子
Bioactive Materials
·
2023
·
IF 18.9
Shenghao Wang,Wenbo He,Huan Wang,Dachuan Liu,Miao Wang,Huilin Yang,Guoqing Pan,Bin Li
Infected bone fractures remain a major clinical challenge for orthopedic surgeons. From a tissue regeneration perspective, biomaterial scaffolds with antibacterial and osteoinductive activities are highly desired, while advanced materials capable of mimicking the pathological microenvironment during the healing process of infected tissues remain an area deserving more research. Hematoma, the gel-like blood coagulum, plays an essential role in bone fracture repair because of its ability to serve as a dynamic and temporary scaffold with cytokines for both pathogen elimination and tissue healing. In light of this, we designed a dynamic hydrogel with hematoma-like antimicrobial or reparative performance for infected bone fracture repair in this study. The proposed dynamic hydrogel network was based on the reversible recognition of a natural glycopeptide antibiotic vancomycin (Van) and its target dipeptide D-Ala-D-Ala (AA), which could serve as a hematoma-like scaffold for obliterating bacteria in the fracture region and promoting bone repair by introducing an endogenous osteogenic peptide (OGP). In vivo experiments demonstrated that the hydrogel could rapidly eradicate bacteria, improve bone regeneration and restore the local inflammatory microenvironment. Together, findings from this study imply that the use of hematoma-like dynamic hydrogel could lead to a biomimetic revolution in surgical strategies against susceptible bone fractures.
研究领域:
生物材料
·
组织工程
·
骨科
·
再生医学
✓ 已验证
DOI
PubMed
高影响因子
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
·
2021
·Vol. 193
·期 7
·2029-2042
·
IF 2.926
Xu Jian-Miao,Cao Hui-Ting,Wang Ming,Ma Bao-Jian,Wang Liu-Yu,Zhang Kai,Cheng Feng,Xue Ya-Ping,Zheng Yu-Guo
d -amino acid oxidase (DAAO) is widely used in the industrial preparation of l -amino acids, and cultivating Escherichia coli ( E. coli ) expressing DAAO for the biosynthesis of l -phosphinothricin ( l -PPT) is very attractive. At present, the biomass production of DAAO by fermentation is still limited in large-scale industrial applications because the expression of DAAO during the fermentation process inhibits the growth of host cells, which limits higher cell density. In this study, the factors that inhibit the growth of bacterial cells during a 5-L fed-batch fermentation process were explored, and the fermentation process was optimized by co-expressing catalase (CAT), by balancing the biomass and the enzyme activity, and by adding exogenous d -alanine ( d -Ala) to relieve the limitation of DAAO on the cells and optimize fermentation. Under optimal conditions, the DO-STAT feeding mode with DO controlled at 30% ± 5% and the addition of 27.5 g/L lactose mixed with 2 g/L d -Ala during induction at 28 °C resulted in the production of 26.03 g dry cell weight (DCW)/L biomass and 390.0 U/g DCW specific activity of DAAO; an increase of 78% and 84%, respectively, compared with the initial fermentation conditions. The fermentation strategy was successfully scale-up to a 5000-L fermenter.
研究领域:
生物技术
·
微生物学
·
酶学
·
工业发酵
✓ 已验证
DOI
PubMed
高影响因子
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