BIOSENSORS & BIOELECTRONICS
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2018
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IF 8.173
Zihua Wang,Yuan Chen,Wenhao Dong,Jiexin Zhou,Bingkai Han,Jun Jiao,Lintao Lan,Peng Miao,Qiang Chen
In this work, carboxylated multi walled carbon nanotubes (CMWCNTs) were firstly prepared and functionalized with poly- L -histidine (PLH), which were then chelated with copper (II) ions to from the nanocomposites of Cu(II)-PLH-CMWCNTs. The nanocomposites could be exploited as an efficient mimic enzyme for sensitive electrochemical detection of salvianic acid A (SAA). Cu(II)-PLH-CMWCNTs owned good charge transfer property and excellent synergetic catalytic effect between the overoxidized imidazole groups and the copper redox-active units. Therefore, highly sensitive electrochemical response to SAA was achieved under optimum experimental conditions. A good linear relationship between differential pulse voltammetry (DPV) peak current and the SAA concentration was established in the range of 0.4–1000 μM. A low detection limit of 0.037 μM and a sensitivity of 0.27 μA μM −1 cm −2 were achieved. The developed biosensor also had advantages of good repeatability, stability and high selectivity, thus, it was successfully applied to the determination of SAA in real samples with satisfactory results, which may have great potential for further exploitation of electroanalysis applications.
研究领域:
生物传感器
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电化学分析
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DOI
PubMed
高影响因子
MOLECULES
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2022
·Vol. 27
·期 18
·6088
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IF 4.927
Ke Shun Hu,Chong Le Chen,Huan Ru Ding,Tian Yu Wang,Qin Zhu,Yi Chen Zhou,Jia Min Chen,Jia Qi Mei,Sheng Hu,Jun Huang,Wei Rui Zhao,Le He Mei
Salvianic acid A (SAA), as the main bioactive component of the traditional Chinese herbSalvia miltiorrhiza, has important application value in the treatment of cardiovascular diseases. In this study, a two-step bioprocess for the preparation of SAA froml-DOPA was developed. In the first step,l-DOPA was transformed to 3,4-dihydroxyphenylalanine (DHPPA) using engineeredEscherichia colicells expressing membrane-bound L-amino acid deaminase fromProteus vulgaris.After that, the unpurified DHPPA was directly converted into SAA by permeabilized recombinantE. colicells co-expressingd-lactate dehydrogenase fromPediococcus acidilacticiand formate dehydrogenase fromMycobacterium vaccaeN10. Under optimized conditions, 48.3 mM of SAA could be prepared from 50 mM ofl-DOPA, with a yield of 96.6%. Therefore, the bioprocess developed here was not only environmentally friendly, but also exhibited excellent production efficiency and, thus, is promising for industrial SAA production.Keywords:salvianic acid A;l-DOPA;membrane-boundl-amino acid deaminases;biocatalysis;molecular biology;biological engineering
研究领域:
生物技术
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药学
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生物化工
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DOI
PubMed
高影响因子
Bioresources and Bioprocessing
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2023
·Vol. 10
·期 1
·1-14
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IF 4.6
Yang Jiahui,Wei Wanqing,Gao Changzheng,Song Wei,Gao Cong,Chen Xiulai,Liu Jia,Guo Liang,Liu Liming,Wu Jing
Salvianic acid A (SAA), used for treating cardiovascular and cerebrovascular diseases, possesses several pharmacological properties. However, the current methods for the enzymatic synthesis of SAA show low efficiency. Here, we constructed a three-enzyme cascade pathway in Escherichia coli BL21 (DE3) to produce SAA from l-dihydroxyphenylalanine (L-DOPA). The phenylpyruvate reductase (LaPPR) from Lactobacillus sp. CGMCC 9967 is a rate-limiting enzyme in this process. Therefore, we employed a mechanism-guided protein engineering strategy to shorten the transfer distances of protons and hydrides, generating an optimal LaPPR mutant, LaPPRMu2 (H89M/H143D/P256C), with a 2.8-fold increase in specific activity and 9.3-time increase in kcat/Km value compared to that of the wild type. Introduction of the mutant LaPPRMu2 into the cascade pathway and the optimization of enzyme levels and transformation conditions allowed the obtainment of the highest SAA titer (82.6 g L−1) ever reported in vivo, good conversion rate (91.3%), excellent ee value (99%) and the highest productivity (6.9 g L−1 h−1) from 90 g L−1 L-DOPA in 12 h. This successful strategy provides a potential new method for the industrial production of SAA.
研究领域:
生物化学
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生物技术
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酶学
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制药
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DOI
PubMed
高影响因子
MOLECULAR PHARMACEUTICS
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2023
·
IF 4.9
Qingfa Tang,Yao Chen,Yusheng Zhang,Kewei Wang,Li Liu,Dongmei Pan,Qiang Liu,Cuiping Jiang
Tanshinone IIA (TS-IIA) and salvianic acid A (SAA) are the main pharmacological active constituents of Danshen, which exhibit potent effects on atherosclerosis. A combination of TS-IIA and SAA might exert a synergistic antiatherosclerotic effect. However, the opposite solubility profiles of TS-IIA and SAA might lead to difficulty in achieving a synergistic combined effect of the two active components. Therefore, in this work, we fabricated a ROS-responsive prodrug micelle for the codelivery of TS-IIA and SAA (TS-IIA-PM) by self-assembling amphiphilic block copolymer PEG5000-SAA/PLA10000-APBA. The amphiphilic polymer was characterized by 1H NMR, FTIR, and alizarin red S competition tests. The ROS responsiveness of TS-IIA-PM was evidenced by time-course monitoring of particle size and morphology changes and drug release behavior in the presence of 1 mM H2O2. We found TS-IIA-PM was stable according to its critical micelle concentration and the unchanged particle sizes in 10% FBS for 7 days. The results of in vitro and in vivo tests revealed that TS-IIA-PM was safe and biocompatible. Furthermore, it was observed that TS-IIA and prodrug micelle could produce synergistic antiatherosclerotic effect based on the results of the antioxidant study, which was further confirmed by a series of pharmocodynamics studies, such as in vitro DiI-oxLDL uptake study, oil red O staining, cholesterol efflux study, inflammatory cytokine analysis, in vivo CD68 immunostaining, and lipid disposition staining studies. Collectively, TS-IIA-PM holds great potential for the safe and efficient codelivery of TS-IIA and SAA for synergistic antiatherosclerosis.
研究领域:
药学
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纳米技术
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心血管疾病
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药物递送系统
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生物医学工程
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DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）