METABOLIC ENGINEERING
·
2016
·
IF 8.201
Yanfen Bai,Hua Yin,Huiping Bi,Yibin Zhuang,Tao Liu,Yanhe Ma
Gastrodin, a phenolic glycoside , is the key ingredient of Gastrodia elata , a notable herbal plant that has been used to treat various conditions in oriental countries for centuries. Gastrodin is extensively used clinically for its sedative, hypnotic, anticonvulsive and neuroprotective properties in China. Gastrodin is usually produced by plant extraction or chemical synthesis, which has many disadvantages. Herein, we report unprecedented microbial synthesis of gastrodin via an artificial pathway. A Nocardia carboxylic acid reductase , endogenous alcohol dehydrogenases and a Rhodiola glycosyltransferase UGT73B6 transformed 4-hydroxybenzoic acid, an intermediate of ubiquinone biosynthesis , into gastrodin in Escherichia coli. Pathway genes were overexpressed to enhance metabolic flux toward precursor 4-hydroxybenzyl alcohol. Furthermore, the catalytic properties of the UGT73B6 toward phenolic alcohols were improved through directed evolution. The finally engineered strain produced 545 mg l −1 gastrodin in 48 h. This work creates a new route to produce gastrodin, instead of plant extractions and chemical synthesis.
研究领域:
生物化学
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合成生物学
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药理学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
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2025
·
IF 3.1
Lin Ge,Yu Xia,Wenxin Xu,Ruobing Jia,Tingting Zhang
Gastrodin, the primary bioactive constituent of Gastrodia elata , possesses numerous remarkable pharmacological properties. In this investigation, UDP-glycosyltransferase from Rauvolfia serpentina (RsUGT) was expressed, subsequently purified and characterized. The maximum yield of the enzyme was 17.57 mU/ml and possessed a relative molecular weight of approximately 77.7 kDa. Utilizing GST affinity resin, RsUGT was purified 20.8-fold, with an overall recovery rate of 58.6% and specific activity of 79.2 mU/mg. The optimal temperature and pH for RsUGT was identified as 40°C and 10.0, respectively. Notably, 2% DMSO could increase the RsUGT activity by 12.15%. The Michaelis-Menten constants K M and V max were determined to be 0.50mM and 171.60 mU/mg. By optimizing the conditions for the enzymatic biosynthesis of gastrodin by RsUGT, the highest gastrodin production was 285.35 mg/l, accompanied by a molar conversion rate of 99.67%. In addition, the conditions of gastrodin biosynthesis by recombinant strain BL-RsUGT were also studied. The highest gastrodin production was 225.99 mg/l, and the corresponding p HBA conversion rate was 98.00%. These findings confirmed the promising potential of RsUGT in the production of gastrodin.
研究领域:
植物学
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药理学
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生物化学
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产品关联
DOI
PubMed
高影响因子
CATALYSIS LETTERS
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2023
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IF 2.8
Xia Hui,Zhang Zihan,Ding Ji,Jiang Kehan,Xue Feng
Gastrodin is an important plant-derived medicinal ingredient that has been used to treat various diseases. Biosynthesis is an effective strategy for gastrodin production, but the expression and activity of natural glycosyltransferases (GTs) is generally a limiting factor. To identify highly efficient GTs, four plant-derived enzymes were identified by homologous sequence analysis and expressed in Escherichia coli BL21 (DE3). A GT from Arabidopsis thaliana (At UGT) displayed the highest activity of 0.45 U/mg, with an optimal pH of 9.0 and high thermostability. Under optimized conditions, 10 mM p -hydroxybenzyl alcohol and uridine diphosphate glucose were converted into 2.67 g/L gastrodin, corresponding to a conversion rate of 94.34% in 12 h. Furthermore, molecular docking simulations were performed between At UGT and p -hydroxybenzyl alcohol, revealing the catalytic mechanism of At UGT. This study provides a novel and effective biocatalyst for the synthesis of gastrodin. Graphical Abstract
研究领域:
植物生物化学
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生物技术
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药物化学
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产品关联
DOI
高影响因子
PROCESS BIOCHEMISTRY
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2012
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IF 2.627
Chao-Jun Pei,Jinhyuk Lee,Yue-Xiu Si,Sangho Oh,Wei-An Xu,Shang-Jun Yin,Guo-Ying Qian,Hong-Yan Han
We studied the inhibitory effect of gastrodin on tyrosinase using inhibition kinetics and computational simulation. Gastrodin reversibly inhibited tyrosinase in a mixed-type manner with K i = 123.8 ± 20.2 mM. Time-interval kinetics revealed the inhibition to be a first-order process with mono- and bi-phasic components. Using AutoDock Vina, we calculated a binding energy of −6.3 kcal/mol for gastrodin and tyrosinase, and we performed a molecular dynamics simulation of the tyrosinase–gastrodin interaction. The simulation results suggested that gastrodin interacts primarily with histidine residues in the active site. A 10-ns molecular dynamics simulation showed that one copper ion in the tyrosinase active site was responsible for the interaction with gastrodin. Our study provides insight into the inhibition of tyrosinase by the hydroxyl groups of gastrodin. A combination of inhibition kinetics and computational calculations may help to confirm the inhibitory action of gastrodin on tyrosinase and define the mechanisms of inhibition.
研究领域:
生物化学
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药物化学
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计算生物学
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DOI
高影响因子
MEDICINAL CHEMISTRY RESEARCH
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2016
·Vol. 26
·期 2
·405-413
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IF 1.436
Wang Jie,Chan Calvin,Huang Feng-wen,Xie Jiang-feng,Xu Hong,Ho Ka-wai,Zheng Sheng-gang,Hu Zhang-li,Lu Jun,He Zhen-dan
The interaction of gastrodin with pepsin has been investigated by enzyme activity assay, fluorescence, UV–Visible, circular dichroism spectra, and molecular docking. The pepsin activity results suggest that gastrodin is an inhibitor of pepsin. The fluorescence experiments show that gastrodin can quench the fluorescence of pepsin via a static quenching process. The thermodynamic analysis suggests that hydrophobic interaction is the main force between pepsin and gastrodin. UV–Visible and circular dichroism spectra studies suggest that the binding of gastrodin leads to a loosening and unfolding of pepsin backbone with partial α-helix being transformed into β-sheet. All these experimental results have been validated by docking studies, which further show that besides hydrophobic interaction, hydrogen bond also help stabilize the gastrodin–pepsin complex. The results reveal the potential to develop the natural compound gastrodin for the treatment of diseases related to the excessive activity of pepsin.
研究领域:
生物化学
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药物化学
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产品关联
DOI
高影响因子
JOURNAL OF FOOD ENGINEERING
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2023
·
IF 5.5
Juanjuan Wen,Ye Tian,Yibin Zhuang,Limin Gou,Wei Ke,Ling Jiang,Minghui Qiu,Yiqun Fan
The issues of flux decline and irreversible membrane fouling must be considered in membrane separation . In this paper, the culture medium, bacterial cells, and proteins were removed using a one-step tight ultrafiltration (UF) membrane separation process. Although the membrane resistance of the tight UF membranes increased owing to their smaller pore size, the stable flux of the 6 nm membrane during fermentation broth filtration of gastrodin was 28.0% higher than that of loose UF membranes, and the flux recovery ratio reached 97.3%. During purification, a higher filtration flux was obtained by retaining the operating pressure within the range of 2–3 bar and the operating temperature range of 50–60 °C. The average flux of a 6 nm 19-channel membrane reached 56 L·m −2 ·h −1 , and the total yield of gastrodin reached 96.2%. This study provides a basis for the large-scale application of tight UF membrane in food industry.
研究领域:
化学工程
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生物技术
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膜技术
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制药学
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食品科学
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产品关联
DOI
高影响因子
JOURNAL OF CHROMATOGRAPHY A
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2014
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IF 4.258
Wenhua Ji,Lingxiao Chen,Xiuli Ma,Xiao Wang,Qianshan Gao,Yanling Geng,Luqi Huang
Molecular imprinted polymers (MIPs) with high selectivity and affinity to gastrodin in water were designed using allyl 2,3,4,6-tetra- O -acetyl-glucopyranoside (TAGL) and 1,2,3,4,5-pentafluoro-6-vinylbenzene (PFVB) as novel functional monomers. Binding characterization of pre-polymerization complexes was researched by nuclear magnetic resonance (NMR) and the MIPs were characterized by scanning electron microscopy (SEM). The properties involving adsorption isotherm, adsorption kinetics and selective recognition capacity were evaluated. The MIPs/TAGL exhibited good site accessibility in which it only took 30 min to achieve adsorption equilibrium and highly selective recognition for the template. Furthermore, the performance of the MIPs/TAGL as solid phase extraction material was investigated in detail and hot water at 50 °C served as the eluting solvent. Pure gastrodin with the recovery of 76.6% was obtained from the aqueous extract of Gastrodia elata roots.
研究领域:
药学
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分析化学
✓ 已验证
DOI
高影响因子
METABOLIC ENGINEERING
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2024
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IF 6.8
Yang Gu,Yaru Jiang,Changfan Li,Jiang Zhu,Xueyao Lu,Jianyue Ge,Mengchen Hu,Jieying Deng,Jingbo Ma,Zhiliang Yang,Xiaoman Sun,Feng Xue,Guocheng Du,Peng Xu,He Huang
Gastrodin, a phenolic glycoside, is a prominent component of Gastrodia elata , which is renowned for its sedative, hypnotic, anticonvulsant, and neuroprotective activities. Engineering heterologous production of plant natural products in microbial host represents a safe, cost-effective, and scalable alternative to plant extraction. Here, we present the construction of an engineered Yarrowia lipolytica yeast that achieves a high-titer production of gastrodin. We systematically refactored the yeast genome by enhancing the flux of the shikimate pathway and optimizing the glucosyl transfer system. We introduced more than five dozen of genetic modifications onto the yeast genome, including enzyme screening, alleviation of rate-limiting steps, promoter selection, genomic integration site optimization, downregulation of competing pathways, and elimination of gastrodin degradation. Meanwhile, we developed a Copper-induced Antisense-Transcriptional Regulation (CATR) tool. The developed CATR toolkit achieved dynamic repression and activation of violacein synthesis through the addition of copper in Y. lipolytica . This strategy was further used to dynamically regulate the pyruvate kinase node to effectively redirect glycolytic flux towards the shikimate pathway while maintaining cell growth at proper rate. Taken together, these efforts resulted in 9477.1 mg/L of gastrodin in shaking flaks and 13.4 g/L of gastrodin with a yield of 0.149 g/g glucose in a 5-L bioreactor, highlighting the potential for large-scale and sustainable production of gastrodin from microbial fermentation.
研究领域:
生物技术
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代谢工程
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合成生物学
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制药
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产品关联
DOI
PubMed
高影响因子
Bioactive Materials
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2021
·
IF 14.593
Hongcai Yang,Qing Li,Limei Li,Shaochun Chen,Yu Zhao,Yingrui Hu,Lu wang,Xiaoqian Lan,Lianmei Zhong,Di Lu
Peripheral nerve regeneration and functional recovery remain a major clinical challenge. Nerve guidance conduit (NGC) that can regulate biological behavior of Schwann cells (SCs) and facilitate axonal regeneration through microenvironmental remodeling is beneficial for nerve regeneration and functional recovery. Gastrodin, a main constituent of a Chinese traditional herbal medicine, has been known to display several biological and pharmacological properties, especially antioxidative, anti‐inflammatory and nerve regeneration. Herein, polyurethane (PU) NGCs modified by different weight ratio of Gastrodin (0, 1 and 5 wt%) were designed for sequential and sustainable drug release, that created a favorable microenvironment for nerve regeneration. The scaffold showed suitable pore structure and biocompatibility in vitro , and evidently promoted morphological and functional recovery of regenerated sciatic nerves in vivo . Compared to the PU and 1%Gastrodin/PU scaffolds, the 5%Gastrodin/PU significantly enhanced the proliferation, migration and myelination of SCs and up-regulated expression of neurotrophic factors, as well as induction of the differentiation of PC12 cells. Interestingly, the obvious anti-inflammatory response was observed in 5%Gastrodin/PU by reduced expression of TNF-α and iNOS, which also evidenced by the few fibrous capsule formation in the subcutaneous implantation. Such a construct presented a similarity to autograft in vivo repairing a 10 mm sciatic nerve defects. It was able to not only boost the regenerated area of nerve and microvascular network, but also facilitate functional axons growth and remyelination, leading to highly improved functional restoration. These findings demonstrate that the 5%Gastrodin/PU NGC efficiently promotes nerve regeneration, indicating their potential for use in peripheral nerve regeneration applications.
研究领域:
生物医学工程
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神经科学
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药理学
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组织工程
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF BIOTECHNOLOGY
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2026
·
IF 4
Lin Ge,Ruobing Jia,Wenxin Xu,Yu Xia,Jianjun Pei
A glycosyltransferase for efficiently converting p HBA into gastrodin was screened. • Enhancing UDP-glucose supply can significantly increase the gastrodin titer in E.coli . • Combinatorial optimization achieved 4578.03 mg/L gastrodin in engineered E. coli . Gastrodin is the principal bioactive component of Gastrodia elata , which is famous for its many remarkable pharmacological properties. In this study, several UDP-glycosyltransferases from different sources were used to evaluate and compare the biotransformation ability of p -hydroxybenzyl alcohol to gastrodin. The recombinant strain BL-RrUGT, expressing the UDP-glycosyltransferase (RrUGT) from Rhodiola rosea , exhibited the highest initial titer of 210.60 mg/L. To further augment production, we enhanced the intracellular UDP-glucose level in Escherichia coli ( E. coli ) K12 by overexpressing two pivotal biosynthetic enzymes: phosphoglucomutase and UDP-glucose pyrophosphorylase. This metabolic engineering strategy yielded the recombinant strain BL-Rr-PG, which boosted the gastrodin titer to 342.40 mg/L. Subsequently, the culture medium and fermentation conditions were optimized. This resulted in a maximum gastrodin titer of 4578.03 mg/L, with a specific productivity of 69.36 mg/L/h and a molar conversion rate of 82.72%. This output represents the highest yield of gastrodin reported in E. coli to date. Download: Download high-res image (198KB) Download: Download full-size image
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DOI
PubMed
高影响因子
JOURNAL OF FOOD COMPOSITION AND ANALYSIS
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2024
·
IF 4
Jinfang Ma,Fengjie Xu,Xue Zhou,Hai Yin,Hui Wang,Caiyun Wang,Jiaze Chen,Baiheng Xie,Fahuan Ge,Furong Huang
Gastrodia elata Blume (Tianma) is widely cultivated and consumed as a food and herbal material in China. With its increasing use as a functional food, Tianma is gaining popularity in the market. However, during the cultivation process, the harvest time affects its maturity and storage time, while the total content of gastrodin (GAS) and p -hydroxybenzyl alcohol (HA) affects its nutritional value , making it difficult to control the quality of Tianma. In this study, a combination of visible-near-infrared (Vis-NIR) spectroscopy and chemometrics was used to identify the harvest time and determine the quality indicator content. A Vis-NIRRamanNet modeling method based on the architecture of RamanNet was proposed and compared with harvest time discrimination models constructed using partial least squares discriminant analysis (PLS-DA), k-nearest neighbor (KNN), support vector machine (SVM), and one-dimensional convolutional neural network (1D-CNN). The results demonstrated that Vis-NIRRamanNet exhibited superior performance, achieving accuracy rates of 0.995 and 0.973 on the training and testing sets, respectively. Subsequently, Vis-NIR spectroscopy combined with support vector regression (SVR) was employed for quantitative analysis of quality indicator content. Wavelength selection methods such as uninformative variable elimination (UVE), genetic algorithm (GA), and competitive adaptive reweighted sampling (CARS) were utilized to optimize the model. The results indicated that the CARS-SVR model provided the best prediction of quality indicator content, with root mean square error of prediction (RMSEP) and coefficient of determination ( R p 2 ) values of 0.257 and 0.901, respectively. Therefore, the combination of Vis-NIR spectroscopy and chemometrics shows significant potential in assisting the selection of high-quality Tianma.
研究领域:
植物学
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食品科学
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药用植物研究
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产品关联
DOI
高影响因子
JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART A
·
2020
·Vol. 108
·期 8
·1713-1725
·
IF 3.525
Qing Li,Limei Li,Mali Yu,Meng Zheng,Yao Li,Jian Yang,Min Dai,Lianmei Zhong,Lin Sun,Di Lu
The extracellular matrix provides cells with a support structure and an attachment site in actual substrate. Its biochemical and surface properties play an important role in and have significant impact on cell attachment, proliferation, migration, differentiation, and gene expression. Leveraging the hydrophilicity and neuroprotective of gastrodin, a gastrodin/polyurethane (PU) elastomer was developed utilizing in situ polymerization and salt-leaching methods. The results showed that gastrodin/PU film had a good flexibility and supporting strength, as well as hydrophilicity. Thus film possessed highly surface area, interconnected porous structure with a pore size (10~60 μm) for cell attachment, and could provide surface cues to augment neurite extension. For PC12 cells cultured within the films, especially the 5gastrodin/PU group, presented a progressive increase with time, coupled with the upregulation of brain-derived neurotrophic factor and glial cell derived neurotrophic factor expression. This is the first report on the construction of a gastrodin/PU porous film, and the results reveal its promise as a scaffold material for neural tissue engineering.
研究领域:
生物材料学
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神经科学
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组织工程学
✓ 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）