BIOSENSORS & BIOELECTRONICS
·
2019
·
IF 9.518
Tingting Tian,Mingxia Liu,Lixia Chen,Fengjiao Zhang,Xin Yao,Hong Zhao,Xiangjun Li
For D-amino acid (DAA) electrochemical biosensors , it is necessary to achieve chiral recognition in racemic solutions or mixtures. However, common chiral recognition is only performed in a single isomer solution. Here, D-amino acid oxidase (DAAO) was used as a chiral selector, and carbon nanotubes (CNTs) as a signal amplifier to construct a non-mediator-style DAA biosensor. The biosensor showed high performance against enantiomer interference: in alanine (Ala) enantiomer mixtures, accurate quantification of D-Ala was achieved when the concentration ratio of L-Ala to D-Ala was 100. In Ala racemic solutions, the linear equation slope was almost consistent with that of standard D-Ala. This high performance was due to the combination of stereoselectivity (enzyme protein) and a catalytic reaction (redox center). The mechanism for the electrical signal change of the biosensor was explored and verified by cyclic voltammetry (CV). The results showed that (i) flavin adenine dinucleotide (FAD, redox center of DAAO) was a direct electroactive substance that produced a reduction peak current; in the presence of O 2 , the amount of FAD increased leading to an increase of the reduction peak current. (ii) In the presence of DAA, the chemical reaction FAD + DAA → imino acids + FADH 2 occurred and consumed FAD, which resulted in its decrease; thus, the reduction peak current also decreased. Under the same oxygen concentration, the linear decrease of the reduction peak current in the presence of DAA was due to FAD consumption. The biosensor was used for practical analyses in milk and urine samples with satisfactory results.
研究领域:
生物化学
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分析化学
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DOI
高影响因子
EUROPEAN FOOD RESEARCH AND TECHNOLOGY
·
2023
·Vol. 249
·期 4
·1137-1146
·
IF 3.3
Zhu Yan,Di Lei,Cheng Shengyu,Jin Yueying,Zhang Minghui,Zhang Lingli,Wang Xin,Li Xi-Ling,Han Yu,Xu Chun-Yan,Min Jun Zhe
A novel UHPLC–MS/MS method with high sensitivity and selectivity for simultaneous detection and chiral resolution of DL-amino acids in beer was established using the newly developed mass spectrometry probe (S)-(3-(4-carboxythiazolidin-3-yl)-3-oxopropyl) diphenylsulfonium (CTOD). The gradient analysis of five chiral amino acids (DL-Asp, DL-Glu, DL-Ala, DL-Pro, and DL-Phe) was carried out on a BEH C18 (2.1 × 150 mm, 1.7 μm) for 30 min with a mobile phase of 0.1% formic acid aqueous solution and 0.1% formic acid methanol/acetonitrile (7:3) solution. The resolutions of DL-amino acids ranged from 1.71 to 5.42. The correlation coefficients were ≥ 0.9994 in the range of 0.25 to 1200 pmol. Meanwhile, limits of detection (LOD) and limits of quantitative (LOQ) analysis were 0.05–0.25 pmol and 0.13–0.50 pmol, respectively. Furthermore, the intra-day and inter-day precision ranged from 0.34% to 6.31%, and the average recoveries ranged from 97.50% to 107.50%. The method was applied to the quantitative analysis and comparison of DL-amino acids in beer samples from 10 different manufacturers, and it was found that the beers were rich in D-amino acids. Four D-AAs including D-Ala, D-Pro, D-Glu, D-Phe (1.9–29.8 μmol L −1 ) and five L-AAs including L-Pro, L-Ala, L-Phe, L-Glu, L-Asp (5.3–2315.9 μmol L −1 ) were detected in the beer samples. The average content of L-AAs in the 10 beer samples was significantly higher than that of D-AAs. This study provides a new method for detecting chiral amino acids in beer and reliable data for evaluating the nutritional value of beer.
Graphical abstract
研究领域:
分析化学
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食品科学
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营养学
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产品关联
DOI
高影响因子
NEW JOURNAL OF CHEMISTRY
·
2022
·Vol. 46
·期 17
·8029-8036
·
IF 3.925
Qingju Liu,Saixue Tang,Hui Wang,Ping Han
The current luminescent metal–organic framework (MOF) based fluorescence detection mostly focuses on achiral molecules. The stereospecific recognition and determination of MOF-based detection remain challenging. A novel luminescent terbium-based MOF (Tb-MOF) based biosensing system with excellent performance for stereoselective detection of D-amino acids (D-AAs) was introduced in this paper. D-Amino acid oxidase catalyzes stereospecific oxidative deamination of D-AAs, producing H2O2. The generated H2O2 effectively quenches the fluorescence of Tb-MOF. Under optimum conditions, the D-AAs were detected with excellent efficiency. The linear working range was 0.5–100 μM for D-Alanine (D-Ala) and 0. 5–5 μM for D-Proline (D-Pro), respectively. The limit of detection (Ksv detection) for D-Ala and D-Pro was 0.09 μM and 0.14 μM, respectively. The detection can be performed within 30 min with high selectivity and long-term stability. The fluorescent sensing platform required simple fabrication and operation and was free from complicated reaction processes and expensive reagents. The present sensing system was successfully applied in D-AA detection in human serum with good recovery, indicating its potential application in detection in biological samples.
研究领域:
分析化学
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生物技术
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材料科学
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产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2024
·
IF 8
Qianxi Zhou,Hao Zhang,Tianyu Zeng,Jiahua Yang,Qizhi Liang,Hai Shi,Yongmei Yin,Genxi Li
The dynamic variation of D-amino acids (D-AAs) in human saliva has great potential as the indicator for individual physiological states and disease progression; however, enantiospecific profiling of D-AAs is a great challenge. In this work, a nanoreactor based on mesoporous hydrogen-bonded organic framework (MHOF) has been fabricated for the enantiospecific profiling of D-AAs. The nanoreactor is prepared by using MHOFs as the scaffold to organize the biocatalytic cascade of D-AA oxidase (DAAO) and horseradish peroxidase (HRP). In the meanwhile, the large, stable, and robust transport channels of MHOFs allow the unimpeded transport of the involved biocatalytic substrates. Once D-AA substrates efficiently transport into MHOFs, the chromogenic reactions of 3,3′,5,5′-tetramethylbenzidine (TMB) and H 2 O 2 can be activated. So, using this MHOF nanoreactor, enantiospecific and visual detection of D-AAs can be achieved. Moreover, the stable nanoreactor can be operated in complex solution and utilized for the profiling of D-AA levels in saliva samples collected from gastric cancer patients, suggesting that D-AAs may be a powerful fingerprint for the diagnosis of gastric cancer. Therefore, the fabricated MHOF-based nanoreactor may provide an emerging tool for the enantiospecific profiling of D-AAs and supports the D-AA fingerprint as the indicator of gastric cancer diagnosis.
研究领域:
医学
·
生物化学
·
材料科学
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产品关联
DOI
高影响因子
Analytical Methods
·
2017
·Vol. 9
·期 43
·6131-6138
·
IF 1.9
Xiangyu Wang,Huihui Wu,Rongying Luo,Donghai Xia,Zhengjin Jiang,Hai Han
D-amino acids are currently paid attention to as new physiologically active substances. Foodstuffs and beverages containing D-amino acids are a matter of interest. Until now, the profiles of D-amino acids have not been reported in natural and fermented teas except theanine. In this study, an off-line 2D-HPLC method combining a Gemini C18 column and a CHIRALPAK® IC-3 column or a self-prepared poly(MQD-co-HEMA-co-EDMA) monolithic capillary column was employed for the separation and detection of D-amino acids in tea samples with the co-existence of a large amount of L-amino acids. The free amino acid fractions in longjing, black, oolong, and pu-erh tea samples were separated and collected after pre-column derivatization using 9-fluorenylmethoxycarbonyl (FMOC) chloride in the reversed-phase mode, and then were concentrated and separated to D- and L-forms on a chiral column. Among them, the D-form of isoleucine (Ile) (1.0–1.6%), alanine (Ala) (0.1–0.8%), phenylalanine (Phe) (0.1–0.4%), valine (Val) (0.2–0.3%), serine (Ser) (0.1–0.2%), aspartic acid (Asp) (0.2%), and proline (Pro) (0.1%) were detected in longjing and oolong teas. Differences between natural tea and fermented tea in the profiles of D-amino acids as well as the total amino acids were also observed. The results provided useful information for the bio-function research of D-amino acids in tea.
研究领域:
食品科学
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分析化学
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生物化学
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产品关联
DOI
高影响因子
BIOORGANIC CHEMISTRY
·
2020
·
IF 4.831
Liang Yan,Yongqi Ke,Yuhe Kan,Dao Lin,Jingkui Yang,Yujian He,Li Wu
The isomerization of l -amino acids in peptides and proteins into d -configuration under physiological conditions would affect the physiological dysfunction and caused protein conformational diseases. The presence of d -amino acids might change the higher-order structure of proteins and triggered abnormal aggregation. In order to better understand this phenomenon and promote degradation, we systematically studied the enzymatic hydrolysis of a series of peptides obtained by replacing l -amino acids in different positions of template peptide KYNETWRSED with d -amino acids under the action of Protease K. The results showed that, compared with normal peptide, isomerization of different amino acids had different effects on the anti-enzymatic hydrolysis of the peptides, especially d -tryptophan at position 6, which significantly inhibited enzymatic hydrolysis. The analysis of the peptide cleavage site revealed that the efficiency of enzymatic hydrolysis mainly depended on the isomerization of the amino acids at a specific site of the peptide cleavage. Further studies showed that the enzymatic hydrolysis of substrates could be facilitated by optimized reaction conditions such as temperature, pH, addition of metal ions, and change of buffer. In this way the accumulation of disease-associated d -amino acid containing polypeptides/proteins could be prevented.
研究领域:
生物化学
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分子生物学
·
蛋白质化学
✓ 已验证
DOI
PubMed
高影响因子
BIOORGANIC CHEMISTRY
·
2025
·
IF 4.7
Shuaishuai Cui,Cunxin Guo,Liang Yan,Yujian He,Li Wu
The incorporation of D-amino acids can influence the structure and enzymatic stability of proteins or peptides, especially when they are introduced at enzyme cleavage sites, significantly inhibiting the enzymatic hydrolysis of peptides. The abnormal accumulation of these peptides has been linked to age-related conditions, including cataracts and Alzheimer's disease. N-terminal acetylation, an essential post-translational modification, significantly enhances the physicochemical properties of peptides and plays an essential role in regulating their performance within biological systems. This research examined the impact of N-terminal acetylation on the enzymatic hydrolysis of peptides incorporating D-amino acids. Enzymatic activity assessments showed that N-terminal acetylation greatly promoted the enzymatic breakdown of these peptides by Proteinase K (PROK), with the substrate decay rate constant for the acetylated peptide Ac-6-w increasing by 17.5 times. This enhancement was specific to serine-type proteases, which exhibited a comparable cleavage pattern. Molecular docking further demonstrated that N-terminal acetylation improved interactions within the catalytic triad of serine proteases, leading to faster enzymatic degradation. The results provide novel insights into the impact of N-terminal acetylation on the enzymatic behavior of peptides incorporating D-amino acids, and they propose a potential approach for targeting these peptides to preserve normal physiological functions.
研究领域:
生物化学
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医学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
MOLECULES
·
2025
·Vol. 30
·期 3
·441
·
IF 4.6
Shuaishuai Cui,Zhaoyang Jin,Tonglin Yu,Cunxin Guo,Yujian He,Yuhe Kan,Liang Yan,Li Wu
The accumulation of D-amino acid-containing peptides is associated with age-related diseases such as Alzheimer’s disease and cataracts, while glycosylation is an important modification of proteins and plays a key role in improving the physicochemical properties of peptides and facilitating their regulation in biological systems. This study investigates the effects of glycosylation position, glycan number, and monosaccharide structure on the conformation and enzymatic degradation of D-amino acid-containing peptides, using KYNEtWRSED (5-t) as a model peptide and six monosaccharides as model glycans. The results demonstrated that glycosylation inhibited the enzymatic degradation of 5-t in the presence of most serine-like proteases. However, in the presence of chymotrypsin, glycosylation with modified monosaccharides (except for β-D-GalNAc) promoted the degradation of 5-t. Furthermore, glycosylation had no effect on the cleavage site of 5-t. Molecular docking analysis revealed that the hydrogen bonding and electrostatic interactions between the glycopeptide and chymotrypsin were markedly strengthened, likely serving as a key determinant of the enzymatic effects. Collectively, these findings highlight the potential of glycosylation to enhance the therapeutic and biomedical applications of D-amino acid-containing peptides in disease treatment and drug design.
研究领域:
生物化学
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分子生物学
·
医学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
·
2024
·
IF 2.8
Wenchan Deng,Chundan Ye,Wei Wang,Rongrong Huang,Cheng Guo,Yuanjiang Pan,Cuirong Sun
Colorectal cancer (CRC) is a common malignant tumor in the gastrointestinal tract. Changes in amino acid metabolites have been implicated in tumorigenesis and disease progression. Biomarkers on the basis of chiral amino acids, especially D-amino acids, have not been established for early diagnosis of CRC. Quantification of chiral amino acids, especially very low concentrations of endogenous D-amino acids, is technically challenging. We report here the quantification of L- and D-amino acids in urine samples collected from 115 CRC patients and 155 healthy volunteers, using an improved method. The method of chiral labeling, liquid chromatography, and tandem mass spectrometry enabled separation and detection of 28 amino acids (14 L-amino acids, 13 D-amino acids and Gly). Orthogonal partial least squares discriminant analysis identified 14 targeted variables among these chiral amino acids that distinguished the CRC from the healthy controls. Binary logistic regression analysis revealed that D-α-aminobutyric acid (D-AABA), L-alanine (L-Ala), D-alanine (D-Ala), D-glutamine (D-Gln) and D-serine (D-Ser) could be potential biomarkers for CRC. A receiver operating characteristic curve analysis of combined multi-variables contributed to an area under the curve (AUC) of 0.995 with 98.3 % sensitivity and 96.8 % specificity. A model constructed with D-AABA, D-Ala, D-Gln, and D-Ser achieved an AUC of 0.988, indicating important contributions of D-amino acids to the association with CRC. Further analysis also demonstrated that the metabolic aberration was associated with age and the development of CRC, D-methionine (D-Met) was decreased in CRC patients with age over 50, and D/L-Gln in patients at stage IV was higher than patients at stage I. This study provides the signature of D-amino acids in urine samples and offers a promising strategy for developing non-invasive diagnosis of CRC.
研究领域:
肿瘤学
·
代谢组学
·
医学诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOELECTROCHEMISTRY
·
2025
·
IF 4.5
Tingting Tian,Lixia Chen,Taiguang Li,Xiang Wang,Sen Yang,Huiqun Wang,Yujie Jiang,Xin Yao,Hong Zhao,Dengchao Wang,Xiangjun Li
Electrochemiluminescence (ECL) is highly recommended in chiral recognition. ECL-based chiral sensors highly desire a sensitive sensing interface for signal conversion and absolute chiral discrimination. The ECL emission based on a luminol-dissolved O 2 system received much attention due to its nontoxicity and stability. However, the drawback of weak ECL emission hinders the fast signal conversion from chiral discrimination to ECL response. Herein, the amplification strategy of ECL emission is proposed based on the electrochemical reduction enhanced O 2 reduction reaction (ORR). Cadmium sulfide decorated on carbon-nanotubes (CdS/CNTs) with easy synthesis, wide-pH operation, and suitable valence-conduction band position is employed. Upon cathodic scan, the electrons transfer from electrochemically reduced-CdS/CNTs to O 2 and H 2 O 2 , thus accelerating the generation of reactive oxide species (ROS) and furthering ECL emission. Furthermore, the chiral ECL sensing interface is well-designed by combining the chiral recognition of D-amino acid oxidase (DAAO) with the signal transduction and amplification of CdS/CNTs-enhanced ECL emission. During DAAO-catalyzed enantioselective-oxidations of alanine, the O 2 is converted to H 2 O 2 , which tunes the ROS generation. With synergetic regulations of ROS generation by nano-derived CdS/CNTs and bio-derived DAAO, alanine enantiomers are highly discriminated and the L-alanine is quantitatively detected with the most competitive detection limit so far (0.014 fM).
研究领域:
分析化学
·
生物化学
✓ 已验证
产品关联
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
✓ 已验证
产品关联
DOI
高影响因子
MICROCHEMICAL JOURNAL
·
2024
·
IF 4.9
Shuo Qin,Zongze Li,Boran Zhou,Bin Liu,Yuting Xue,Ruixue Zhao,Lirong Zheng,Zhengbo Chen,Xia Zuo
Overuse of benzoate as a preservative is bad for human health. Therefore, we need to achieve quantitative detection of benzoate. Colorimetric nanozyme sensors have garnered significant interest owing to their exceptional efficacy and affordability. In this article, Fe-N-C dodecahedral nanozymes with superior peroxidase-like catalytic activity are prepared using ZIF-8 as a precursor. Therefore, we construct a colorimetric sensor for benzoate detection with a detection limit of 0.33 μM through integrating the inhibitory impact of benzoate on D-amino acid oxidase activity with the Fe-N-C-catalyzed 3,3′,5,5′-tetramethylbenzidine (TMB)–H 2 O 2 system, and this reaction is completed within 10 min. It is noteworthy that a recovery rate of 96.8–109 % has been attained for the detection of benzoate in beverages.
研究领域:
食品安全
·
生物化学
·
纳米技术
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）