COMBINATORIAL CHEMISTRY & HIGH THROUGHPUT SCREENING
·
2019
·Vol. 22
·期 2
·113-122
·
IF 1.503
Zhang Xin-guo,Ma Guo-Yan,Kou Fei,Liu Wen-Jie,Sun Qiao-Yun,Guo Guang-Jun,Ma Xiao-Di,Guo Si-Jia,Jian-Ning Zhu
Background: Adenosine deaminase (ADA) is an important enzyme in purine metabolism and is known as a potential therapeutic target for the treatment of lymphoproliferative disorders and cancer. Traditional Chinese Herbal Medicine (TCHM) is widely used alone or in combination with chemotherapy
to treat cancer, due to its ability to deliver a broad variety of bioactive secondary metabolites as promising sources of novel organic natural agents. Objective: In the present study, 29 varieties of medicinal plants were screened for the presence of ADA inhibitors. Results: Extracts
from Reynoutria japonica, Glycyrrhiza uralensis, Lithospermum erythrorhizon, Magnolia officinalis, Gardenia jasminoides, Stephania tetrandra, Commiphora myrrha, Raphanus sativus and Corydalis yanhusuo demonstrated strong ADA inhibition with rates greater than 50%. However, Reynoutria japonica
possessed the highest ADA inhibitory activity at 95.26% and so was used in our study for isolating the ADA inhibitor to be further studied. Eight compounds were obtained and their structures were identified. The compound H1 had strong ADA inhibitory activity and was deduced to be emodin by
1H and 13C-NMR spectroscopic analysis with an IC50 of 0.629 mM. The molecular docking data showed that emodin could bind tightly to the active site of ADA. Our results demonstrated that emodin displayed a new biological activity which is ADA inhibitory activity with high cytotoxic activity
against K562 leukemia cells. The bioactivity of cordycepin was significantly increased when used in combination with emodin. Conclusion: Emodin may represent a good candidate anti-cancer therapy and adenosine protective agent.
研究领域:
药理学
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肿瘤学
·
植物化学
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DOI
PubMed
高影响因子
ANALYST
·
2015
·Vol. 140
·期 5
·1572-1577
·
IF 4.107
Fen Cheng,Yue He,Xiao-Jing Xing,Dai-Di Tan,Yi Lin,Dai-Wen Pang,Hong-Wu Tang
A novel strategy for the fabrication of a colorimetric aptasensor using label free gold nanoparticles (AuNPs) is proposed in this work, and the strategy has been employed for the assay of adenosine deaminase (ADA) activity. The aptasensor consists of adenosine (AD) aptamer, AD and AuNPs. The design of the biosensor takes advantage of the special optical properties of AuNPs and the interaction between AuNPs and single-strand DNA. In the absence of ADA, the AuNPs are aggregated and are blue in color under appropriate salt concentration because of the grid structure of an AD aptamer when binding to AD, while in the presence of the analyte, AuNPs remain dispersed with red color under the same concentration of salt owing to ADA converting AD into inosine which has no affinity with the AD aptamer, thus allowing quantitative investigation of ADA activity. The present strategy is simple, cost-effective, selective and sensitive for ADA with a detection limit of 1.526 U L−1, which is about one order of magnitude lower than that previously reported. In addition, a very low concentration of the inhibitor erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA) could generate a distinguishable response. Therefore, the AuNP-based colorimetric biosensor has great potential in the diagnosis of ADA-relevant diseases and drug screening.
研究领域:
生物化学
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纳米技术
·
分析化学
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DOI
PubMed
高影响因子
Cell Chemical Biology
·
2017
·
IF 0
Yongliang Xia,Feifei Luo,Yanfang Shang,Peilin Chen,Yuzhen Lu,Chengshu Wang
Summary Cordycepin (COR) and pentostatin (PTN) are adenosine analogs with related bioactivity profiles as both mimic adenosine and can inhibit some of the processes that are adenosine dependent. Both COR and PTN are also natural products and were originally isolated from the fungus Cordyceps militaris and the bacterium Streptomyces antibioticus , respectively. Here, we report that not only is PTN produced by C. militaris but that biosynthesis of COR is coupled with PTN production by a single gene cluster. We also demonstrate that this coupling is an important point of metabolic regulation where PTN safeguards COR from deamination by inhibiting adenosine deaminase (ADA) activity. ADA is not inhibited until COR reaches self-toxic levels, at which point ADA derepression occurs allowing for detoxification of COR to 3′-deoxyinosine. Finally, we show that using our biosynthetic insights, we can engineer C. militaris to produce higher levels of COR and PTN.
研究领域:
真菌学
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生物化学
·
药物开发
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DOI
BIOSENSORS & BIOELECTRONICS
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2016
·
IF 7.476
Jiaona Xu,Chunying Wei
Two general and reliable fluorescence sensors were proposed in this work utilizing aptamer DNA-templated silver nanoclusters (Ag NCs). Both DNA-AgNCs could be used for label-free detecting of ATP with the limits of detection of 0.44 and 0.65 mM. One of them was further applied to monitor the activity of adenosine deaminase (ADA). In our effort to elucidate the light-up mechanism, we studied a total of six Ag NCs prepared by different DNA sequences , and found that they showed different sensitivity to ATP. Both BT3T3- and BT3T3(R)-templated Ag NCs were chose to make particular studies by UV–vis, TEM, fluorescence, and TCSPC methods. The results showed that when DNA-Ag NCs was kept for 1.5 h and presented a strong fluorescence, the addition of ATP failed to cause a large change of fluorescence intensity ; on the contrary, after Ag NCs was kept for 24 h and emitted a weak fluorescence, adding ATP was able to result in the large fluorescence enhanced of 43 and 33 times for BT3T3- and BT3T3(R)-templated Ag NCs, respectively. The possible mechanism was also suggested that ATP binding to aptamer segment of template induced the change of the DNA secondary structure , which made the aggregated Ag nanoparticles disperse into Ag NCs with an average diameter of about 2 nm that were responsible for the large fluorescence increase. Moreover, ATP could protect the fluorescence intensity of BT3T3(R)-templated Ag NCs from quenching for at least 9 h.
研究领域:
生物化学
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Ping Qiu,Longjian Huang,Jiayi Zhang,Wenbo Li,Xudong Xin,Luchang Chen,Qianli Tang,Kai Zhang,Xianjiu Liao,Jihua Wei
We report the development of a novel split aptamer-driven electrochemical biosensor integrating entropy-driven Signal amplification (EDSA) and CRISPR-Cas12a technology for the highly sensitive and specific detection of adenosine deaminase (ADA). ADA is an important biomarker involved in immune regulation and purine metabolism, and its accurate quantification is critical for diagnosing various diseases. In our system, ADA catalyzes the deamination of adenosine to inosine, resulting in the release of Aptamer1 from a split aptamer structure. The liberated Aptamer1 initiates a strand displacement reaction within a Blocker/Bandage/Scaffold complex, triggering a cascade of entropy-driven amplification cycles that generate a DNA duplex capable of activating CRISPR-Cas12a. Activated Cas12a exhibits trans -cleavage activity, specifically cleaving a ferrocene-labeled single-stranded DNA (ssDNA) probe immobilized on a gold electrode, leading to a measurable decrease in electrochemical signal. The biosensor operates under isothermal and enzyme-free conditions, providing simplicity, rapid detection, and low cost. It achieves a low detection limit of 0.688 U·L −1 with a wide linear range from 1 to 20,000 U·L −1 . The system demonstrates excellent specificity against ADA analogues and common interfering proteins and performs reliably in diluted human serum samples. This strategy represents a versatile and modular platform for point-of-care diagnostics. By altering the aptamer sequence, the biosensor can be adapted for the detection of other clinically relevant proteins and enzymes, contributing to advances in personalized medicine.
研究领域:
生物传感技术
·
分子诊断
·
医学检测
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DOI
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
·
2023
·
IF 3.4
Lu Zhao,Wenhe Zhang,Qi Wang,Huibin Wang,Xiao Gao,Bin Qin,Xian Jia,Song You
L -Phosphinothricin ( L -PPT) is the effective constituent in racemic PPT (a high-efficiency and broad-spectrum herbicide), and the exploitation of green and sustainable synthesis route for L -PPT has always been the focus in pesticide industry. In recent years, “one-pot, two-step” enzyme-mediated cascade strategy is a mainstream pathway to obtain L -PPT. Herein, Rg DAAO and Bs LeuDH were applied to expand “one-pot, two-step” process. Notably, a NADH-dependent leucine dehydrogenase from Bacillus subtilis ( Bs LeuDH) was firstly characterized and attempted to generate L -PPT, achieving an excellent enantioselectivity (99.9% ee ). Meanwhile, a formate dehydrogenase from Pichia pastoris ( Pp FDH) was utilized to implement NADH cofactor regeneration and only CO 2 was by-product. Sufficient amount of the corresponding keto acid precursor PPO was obtained by oxidation of D -PPT relying on a D -amino acid oxidase from Rhodotorula gracilis ( Rg DAAO) with content conversion (46.1%). L -PPT was ultimately prepared from racemized PPT via oxidative deamination catalyzed by Rg DAAO and reductive amination catalyzed by Bs LeuDH, achieving 80.3% overall yield and > 99.9% ee value.
研究领域:
生物化学
·
农药合成
·
酶工程
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DOI
PubMed
高影响因子
NUCLEIC ACIDS RESEARCH
·
2024
·
IF 16.6
Wu Yaokang,Li Yang,Liu Yanfeng,Xiu Xiang,Liu Jiaheng,Zhang Linpei,Li Jianghua,Du Guocheng,Lv Xueqin,Chen Jian,Ledesma-Amaro Rodrigo,Liu Long
Mutagenesis driving genetic diversity is vital for understanding and engineering biological systems. However, the lack of effective methods to generate in-situ mutagenesis in multiple genomic loci combinatorially limits the study of complex biological functions. Here, we design and construct MultiduBE, a dCas12a-based multiplexed dual-function base editor, in an all-in-one plasmid for performing combinatorial in-situ mutagenesis. Two synthetic effectors, duBE-1a and duBE-2b, are created by amalgamating the functionalities of cytosine deaminase (from hAPOBEC3A or hAID*Δ ), adenine deaminase (from TadA9), and crRNA array processing (from dCas12a). Furthermore, introducing the synthetic separator Sp4 minimizes interference in the crRNA array, thereby facilitating multiplexed in-situ mutagenesis in both Escherichia coli and Bacillus subtilis. Guided by the corresponding crRNA arrays, MultiduBE is successfully employed for cell physiology reprogramming and metabolic regulation. A novel mutation conferring streptomycin resistance has been identified in B. subtilis and incorporated into the mutant strains with multiple antibiotic resistance. Moreover, surfactin and riboflavin titers of the combinatorially mutant strains improved by 42% and 15-fold, respectively, compared with the control strains with single gene mutation. Overall, MultiduBE provides a convenient and efficient way to perform multiplexed in-situ mutagenesis.
研究领域:
分子生物学
·
基因工程
·
微生物学
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产品关联
DOI
PubMed
高影响因子
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
·
2024
·
IF 2.5
He Wang,Haidu Yuan,Jianhao Zhang,Wenjing Yan
Bacterial infections are becoming a significant threat to global human health due to the growing prevalence of biofilm-related infections and the rise in antibiotic resistance. D/ l -cysteine functionalized chiral gold nanoparticles (D/P-Au NPs or L/P-Au NPs) have demonstrated a potent antibacterial effect against E. coli , while the mechanism remains to be elucidated through additional research. Threonine deaminase (TD) is a crucial enzyme involved in branched-chain amino acid (BCAA) biosynthesis in E. coli and is involved in cysteine's antimicrobial effects. This study investigated the interaction between chiral Au NPs (D/P-Au NPs or L/P-Au NPs) and TD as well as its effect on enzyme activity. It demonstrates that chiral Au NPs interact with TD through hydrophobic forces, forming a ground state complex that induces changes in the secondary structure of TD and reduces enzyme activity in a concentration-dependent manner. We found that the exogenous supplementation of isoleucine and valine (2 mg/mL) significantly reduced the antibacterial activity of chiral Au NPs, especially for L/P-Au NPs. The proteomics results indicate that the expression of ilvA and ilvB was down-regulated after L/P-Au NPs treatment, which would interfere with the synthesis of BCAAs. These results demonstrate that chiral Au NPs cause cell death of E. coli partly due to inhibition of TD enzyme activity and the synthesis of branched-chain amino acids.
研究领域:
微生物学
·
纳米生物学
·
生物化学
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DOI
PubMed
高影响因子
CARBOHYDRATE POLYMERS
·
2022
·
IF 10.723
Qingxia Yuan,Hong Li,Qian Wang,Shujing Sun,Ziyu Fang,Hao Tang,Xiaohuo Shi,Jing Wen,Lianghua Huang,Meng Bai,Shengping Zhong,Yonghong Liu,Chenghai Gao,Longyan Zhao
Stichopus monotuberculatus is a tropical sea cucumber species and used as a folk medicine and tonic food . In this study, a fucosylated glycosaminoglycan (SmFG), the depolymerized SmFG (dSmFG) and its oligosaccharide fractions were prepared. The SmFG and its depolymerized products were comprised of a chondroitin-sulfate-E backbone, and various sulfated fucose side chains, including an unusual disaccharide side chain connected to the C-3 position of D-glucuronic acid (GlcA) or GlcA-ol. A peeling reaction occurred during the deaminative depolymerization process. The dSmFG and its fractions showed strong anticoagulant activity by selectively inhibiting intrinsic tenase complex, and had no anti-factor IIa, Xa and VIIa activity. The anticoagulant activity reduced with the decrease of molecular weight , and the unusual branch and novel reducing end may enhance the anticoagulant activity. These findings can provide significant information for development and utilization of depolymerized products from SmFG in food and pharmaceutical industries .
研究领域:
海洋生物学
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药理学
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食品科学
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产品关联
DOI
PubMed
高影响因子
BIOTECHNOLOGY LETTERS
·
2022
·Vol. 44
·期 5
·635-642
·
IF 2.716
Liu Jia,Liu Jianmin,Yang Bin,Gao Cong,Song Wei,Hu Guipeng,Liu Liming,Wu Jing
Objectives This study aimed to develop an efficient enzymatic strategy for the industrial production of phenylpyruvate (PPA) from l -phenylpyruvic acid ( l -Phe). Results l -amino acid deaminase from Proteus mirabilis was expressed in Escherichia coli BL21 (DE3) and modified to release product inhibition by employing conformational dynamics engineering. Based on structural analysis, two residues (E145/L341) were identified for reducing interactions between the product and enzyme and increasing flexibility of the protein, thereby facilitating the product release. The mutant M2 E145A/E341A exhibited a 3.84-fold reduction in product inhibition and a 1.35-fold increase in catalytic efficiency in comparison to the wild type. Finally, 81.2 g/L PPA production with a conversion of 99.6% was obtained in a 5-L bioreactor. Conclusions The engineered catalyst can significantly reduce product inhibition and facilitate the effective industrial synthesis of PPA.
研究领域:
生物化学
·
工业微生物学
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DOI
PubMed
高影响因子
Plant Signaling & Behavior
·
2022
·
IF 2.734
Xiaomin Wang,Zengyuan Tian,Yu Xi,Yuqi Guo
Seashore mallow (Kosteletzkya virginica), as a noninvasive perennial halophytic oilseed-producing dicot, is native from the Gulf to the Atlantic coasts of the U.S. The purpose of our research was to investigate 1-aminocyclopropane-1carboxylic acid deaminase (ACCD) producing endophytic fungi from K.virginica. A total of 59 endophytic fungal strains, isolated from roots in K.virginica of seedlings, were grouped into 12 genera including in Penicillium, Aspergillus, Fusarium, Trichoderma, Rhizopycnis sp., Ceriporia Donk, Trametes sp., Schizophyllum commune sp., Alternaria, Cladosporium, Cylindrocarpon, and Scytalidium according to sequences of ITS. The ACD activity of 10 endophytic fungi isolated was detected. T.asperellum had the highest ACC deaminase activity among all 10 isolated genera of fungal strains, followed by T. viride. Dry weight and fresh weight of plant, plant height, root length, SOD activity, and chlorophyll content of wheat and soybean inoculated with T.asperellum or T. viride was increased compared with non-inoculated control plants under non salt or salt stress. Further analysis showed that T.asperellum or T.viride strains induced downregulation of the expression of ethylene synthesis-related genes such as ACC oxidase (ACO) and ACC synthase (ACS), thereby reducing ethylene synthesis and damage to plants under salt stress. These endophytic fungi can be used as alternative bioinoculants to increase crop yield in saline soil.
研究领域:
植物科学
·
微生物学
·
农业生物技术
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产品关联
DOI
PubMed
高影响因子
Marine Drugs
·
2020
·Vol. 18
·期 12
·610
·
IF 4.073
Yanzuo Le,Xie Zhou,Jiawen Zheng,Fangmiao Yu,Yunping Tang,Zuisu Yang,Guofang Ding,Yan Chen
This study was designed to investigate the effects and underlying mechanisms of Astaxanthin (AST) on high-fructose-induced hyperuricemia (HUA) from the perspectives of the uric acid (UA) synthesis and excretion in rat models. Following six weeks of a 10% fructose diet, the level of serum UA effectively decreased in the AST groups as compared to the model group. The enzymatic activities of xanthine oxidase (XOD) and adenosine deaminase (ADA) were significantly inhibited, and the mRNA expression levels of XOD and ADA significantly decreased after the AST administration. These results suggested that the AST reduced UA synthesis by inhibiting the mRNA expressions and enzyme activities of XOD and ADA, thereby contributing to HUA improvement. On the hand, the relative expressions of the mRNA and protein of kidney reabsorption transport proteins (GLUT9 and URAT1) were significantly down-regulated by AST, while that of the kidney secretion proteins (OAT1, OAT3 and ABCG2) were significantly up-regulated by AST. These results indicated that the AST promoted UA excretion by regulating the urate transport proteins, and thus alleviated HUA. This study suggested that the AST could serve as an effective alternative to traditional medicinal drugs for the prevention of fructose-induced HUA.Keywords:astaxanthin;fructose;hyperuricemia;adenosine deaminase;xanthine oxidase;urate transporters
研究领域:
药理学
·
生物化学
·
营养学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）