Molecular Catalysis
·
2024
·
IF 3.9
Yuqiu Wu,Fanjin Meng,Jiaolong Fu,Yuyang Zhou,Huan Fan,Xin Ju,Liangzhi Li
D-xylose, a biomass derivative, is used as a substrate to generate D-xylulose and D-ribulose with high-added value through a cascade reaction. During this process, the thermal stability and catalytic efficiency of rate-limiting enzymes during cascade reactions are important issues that must be addressed in industrial applications. In this study, we prepared and validated PDA/SiO 2 microspheres as highly loaded immobilization carriers by oxidative self-polymerization of polydopamine (PDA) on silica (SiO 2 ) microspheres. D-xylose was successfully catalyzed by the co-immobilized D-5-phosphate ribose isomerase B (RpiB) and D-tagatose-3-epimerase (DTE), denoted PDA/SiO 2 -RpiB-DTE, to produce D-ribulose and D-xylulose. After immobilization, the K m of co-immobilized RpiB and DTE decreased, which suggested increased substrate affinity. The half-life of the co-immobilized enzymes was 3.5–8 fold higher than that of the free enzymes at 50–70 °C, and the inactivation rate constant was reduced, demonstrating that immobilization improved significantly the thermal stability. The conversion of D-xylose at 10 mM was 29.98% under optimal conditions, which contained 9.85% D-ribulose and 20.13% D-xylulose. PDA/SiO 2 -RpiB-DTE showed up to 89.10% initial enzyme activity after 3 recycle runs. The thermal stability and recovery aspects were improved by immobilization, which facilitates the use of biomass to produce high-value-added products and further exploitation.
研究领域:
生物技术
·
酶工程
·
生化工程
✓ 已验证
产品关联
DOI
高影响因子
CHEMICAL ENGINEERING SCIENCE
·
2021
·
IF 4.311
Xiaoyan Chen,Fan Wang,Qiang Yu,Shijie Liu,Wen Wang,Yu Zhang,Zhongming Wang,Zhenhong Yuan
One pot cascade of enzymatic catalysis was applied for d -allulose synthesis from d -glucose by the combination of d -glucose isomerase (GI) from Escherichia coli MG1655 and d -allulose 3-epimerase (DAE) from Agrobacterium tumefaciens C58. The synthesis process was investigated by characterizing the initial substrate concentration, molar ratios of GI and DAE, the enzyme loading, and contents of cobalt and magnesium ions. The increase of enzyme loading and the molar ratios of GI vs DAE caused the raising of d -allulose yield from 4% to 9%, verifying that the generation rate of d -fructose was an essential factor for rare sugar conversion improvement. The optimal enzyme loadings required for reactions with substrate concentrations of 100 g L −1 and 200 g L −1 were 60 U and 200 U. Kinetics stimulation was conducted to reveal the underlying relationship within the three sugars converted by dual-enzyme, with the fitting curve validated the reliability of the model.
研究领域:
生物化学
·
酶工程
·
食品科学
✓ 已验证
产品关联
DOI
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
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Frontiers in Bioengineering and Biotechnology
·
2022
·
IF 6.064
Wang Jing,Chen Qiaoyu,Wang Xin,Chen Kequan,Ouyang Pingkai
D-1,2,4-Butanetriol (BT) has attracted much attention for its various applications in energetic materials and the pharmaceutical industry. Here, a synthetic pathway for the biosynthesis of BT from d-arabinose was constructed and optimized in Escherichia coli. First, E. coli Trans1-T1 was selected for the synthesis of BT. Considering the different performance of the enzymes from different organisms when expressed in E. coli, the synthetic pathway was optimized. After screening two d-arabinose dehydrogenases (ARAs), two d-arabinonate dehydratases (ADs), four 2-keto acid decarboxylases (ADXs), and three aldehyde reductases (ALRs), ADG from Burkholderia sp., AraD from Sulfolobus solfataricus, KivD from Lactococcus lactis IFPL730, and AdhP from E. coli were selected for the bio-production of BT. After 48 h of catalysis, 0.88 g/L BT was produced by the recombinant strain BT5. Once the enzymes were selected for the pathway, metabolic engineering strategy was conducted for further improvement. The final strain BT5ΔyiaEΔycdWΔyagE produced 1.13 g/L BT after catalyzing for 48 h. Finally, the fermentation conditions and characteristics of BT5ΔyiaEΔycdWΔyagE were also evaluated, and then 2.24 g/L BT was obtained after 48 h of catalysis under the optimized conditions. Our work was the first report on the biosynthesis of BT from d-arabinose which provided a potential for the large-scale production of d-glucose-based BT.
研究领域:
生物化学
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微生物学
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合成生物学
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产品关联
DOI
PubMed
高影响因子
AMB Express
·
2019
·Vol. 9
·期 1
·1-11
·
IF 2.226
Guo Zongren,Long Liangkun,Ding Shaojun
d -Mannose and l -ribose are two important monosaccharides, which have attracted public attention recently because of their great application potentials in food, cosmetic and pharmaceutical industries. Sugar isomerases catalyze the sugar isomerization and therefore can be used as the biocatalysts for production of the high-value sugars from inexpensive sugars. l -arabinose isomerase catalyzes the conversion of l -arabinose to l -ribulose, while d -lyxose isomerase catalyzes l -ribulose and d -fructose to l -ribose and d -mannose, respectively. In this paper, a putative d -LI from Bacillus velezensis (BvLI) was identified, characterized and used to produce d -mannose and l -ribose from d -fructose and l -arabinose, respectively. The recombinant BvLI exhibited a maximum activity at 55 °C and pH 6.5, in the presence of 0.1 mM Co 2+ . Approximately 110.75 g/L d -mannose was obtained from 500 g/L d -fructose in 6 h by the recombinant BvLI, and approximately 105 g/L l -ribose was obtained from 500 g/L l -arabinose in 8 h by the successive biocatalysis of l -arabinose isomerase from Bacillus licheniformis (BlAI) and BvLI.
研究领域:
生物化学
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生物技术
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食品科学
·
制药学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Molecular Catalysis
·
2022
·
IF 5.089
Hengtao Tang,Zheng Zhou,Zhi Chen,Xin Ju,Liangzhi Li
Monosaccharides from biomass can be used as cost-effective raw materials to produce rare sugars which are valuable sweeteners. Enzymes play a pivotal role in such biomass biorefinery processes to develop novel synthetic routes, and improve the environmental friendliness of current routes. Here we propose a proof of concept biorefinery process to prepare useful rare sugars of D-xylulose and D-ribulose from D-xylose which was abundant in biomass. Ribose-5-phosphate isomerase B from Ochrobactrum sp. (OsRpiB) with a unique substrate scope was engineered to intensify the activity of transforming D-xylose to D-xylulose. A double mutant R94N/N137Q with 80% higher catalytic efficiency was identified, and kinetic and molecular dynamic studies were conducted to explore the mechanism underneath. The results revealed a correlation between enhancing stability and activity simultaneously. Then a one-pot enzyme cascade was constructed with OsRpiB and D-tagatose-3-epimerase from Pseudomonas cichorii and transformed D-xylulose further to D-ribulose. The conversions of D-xylose and D-xylulose in cascade reaction were 1.80-fold and 1.83-fold of that in separate reactions with the aid of process parameter adjustment. The presented work is the first reported enzyme cascade converting D-xylose to D-ribulose, and proves the promising role of monosaccharide isomerase platforms in the refinery of biomass.
研究领域:
生物化学
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酶工程
·
生物质精炼
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产品关联
DOI
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
ENZYME AND MICROBIAL TECHNOLOGY
·
2022
·
IF 3.705
Lei Wang,Kecai Chen,Peng Zheng,Xiang Huo,Fei Liao,Liping Zhu,Meirong Hu,Yong Tao
D -Psicose, a new-generation sugar substitute, has been enzymatically synthesized through D -fructose isomerization. However, isomerization often causes low yields due to unfavorable thermodynamic equilibria, which limited its further industrial application. In this study, we present a redox-driven multi-enzyme cascade, two-step biotransformation system to produce D -psicose from D -fructose. Compared to D -fructose isomerization, this method has a maximized theoretical conversion rate of 100%. D -Psicose-3-epimerase from Clostridiales (CBDPE), ribitol 2-dehydrogenase from Providencia alcalifaciens (PRDH), and formate dehydrogenase from Starkeya (SFDH) were co-expressed in Escherichia coli in the first step to produce D -allitol from D -fructose. Afterward, NADH oxidase from Streptococcus pyogenes (SPNOX), and ribitol 2-dehydrogenase from Rubrivivax sp. (RSRDH) were co-expressed in E. coli to oxidize D -allitol into D -psicose in the second step. The two-step biotransformation system was optimized to maximize the D -fructose-to- D -psicose conversion rate (up to 90%), corresponding to a concentration of 450 mM. This study suggests that this redox-driven multi-enzyme cascade strategy through a sugar-to-alcohol-to-sugar pathway has the advantage of great application for enhanced production of D -psicose and other rare sugars.
研究领域:
生物化学
·
生物技术
·
酶学
·
代谢工程
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产品关联
DOI
PubMed
高影响因子
TOXICOLOGY AND APPLIED PHARMACOLOGY
·
2018
·
IF 3.616
Guolin Zhao,Muhanad Elhafiz,Jingwei Jiang,Debanjan Das,Zhijian Li,Wang Zhou,Sisi Fan,Changling Wang,Ziqiao Yuan,Dengqiu Xu,Zhenzhou Jiang,Luyong Zhang,Tao Wang
8-methoxypsoralen (8-MOP) with ultraviolet A radiation therapy (PUVA) is the standard therapy for patients with psoriasis , despite the reported potential risks of 8-MOP-induced cholestatic liver injury in both humans and animals. Usually, patients with chronic cholestasis exhibit lower serum 25-hydroxy vitamin D (25(OH)D) levels. But those patients receiving PUVA for psoriasis showed an increase in serum 25(OH)D levels, probably highlighting that the vitamin D–vitamin D nuclear receptor (VD-VDR) axis play a protective role in 8-MOP-induced hepatotoxicity. The present study confirmed 8-MOP could increase serum 25(OH)D levels in conventional lighting and diet (CLD) and vitamin D deficient (VDD) Sprague-Dawley rats. Potential liver risks were also found in CLD and VDD rats after 8-MOP treatment. We proved that 8-MOP could be a potent ligand for VDR using molecular docking and luciferase report assay. Effect of 8-MOP on VDR subcellular distribution was determined using human liver cell line L02. We found 8-MOP could increase VDR protein expression in the nuclear and cytosol extracts and also total cell extracts in L02. siRNAs for VDR were used to determine the role of VDR in protecting 8-MOP-induced cholestasis and potential cellular mechanisms. The results showed 8-MOP could affect the CYP7A1 , SHP and MRP3 expression via VDR, and such effects could be reversed by knockdown of VDR expression, suggesting a vital role of VDR involved in 8-MOP-regulated bile acid synthesis and transportation. In conclusion, these results revealed activation of VD–VDR axis may play a beneficial role in 8-MOP-mediated regulation of bile acid synthesis and transportation.
研究领域:
药理学
·
毒理学
·
分子生物学
·
肝病学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Systems Microbiology and Biomanufacturing
·
2023
·
IF 0
Zhang Landong,Gao Changzheng,Song Wei,Wei Wanqing,Gao Cong,Chen Xiulai,Liu Jia,Liu Liming,Wu Jing
D -p -hydroxyphenylglycine (D-HPG) is an important intermediate in the pharmaceutical industry, and it is commonly synthesized by cascading D-hydantoinase (DHase) and D-carbamoylase (DCase). In this study, the stability of DCase was identified as the main problem that limits its application. Therefore, the complexed structure of Ak DCase (DCase from the Agrobacterium sp . strain KNK712) with the substrate N-carbamoyl-D- p -hydroxyphenylglycine (C p HPG) (with 2.52 Å resolution) and catalytic mechanism were resolved. Based on the catalytic mechanism and electrostatic stabilization, salt bridge engineering was adopted to improve Ak DCase thermostability. The best variant, Ak DCase D30A , increased the T m by 2.91 °C and half-life (t 1/2 ) at 40 and 60 °C by 18.43 h and 23.21 min, respectively. After Ak DCase D30A was assembled with Gs DHase (DHase from Geobacillus stearothermophilus SD-1) in a single Escherichia coli cell, the recombinant strain could produce 29.53 g/L D-HPG within 12 h, with a 97% conversion and a 2.46 g/(L·h) space–time yield (STY). The titer of D-HPG increased by 40.55% compared to the E. coli cell harboring pETduet-1- Ak DCase- Gs DHase. The recombinant strain could be used for two cycles. Our research provides a basis for the industrial production of D-HPG.
研究领域:
生物催化
·
蛋白质工程
·
工业微生物学
·
结构生物学
✓ 已验证
DOI
LUMINESCENCE
·
2019
·Vol. 34
·期 7
·767-773
·
IF 1.691
Chunlei Ma,Wensheng Ren,Jianke Tang,Xiaoli Wang,Dongxin Ji,Rongqian Meng,Cuihong Zhang,Qi Wang
Taking advantage of the compelling properties of d -penicillamine ( d -PA) combined with copper, a method for the sensitive and selective determination of d -PA was established using copper nanocluster (Cu NC)-based fluorescence enhancement. d -PA molecules containing a thiol compound showed a strong tendency to combine with the surface of Cu NCs, causing the re-dispersion of nanoclusters and therefore fluorescence intensity was enhanced. Fluorescence enhancement efficiency of Cu NCs induced by d -PA was linear, with the d -PA concentration varying from 0.6–30 μg ml −1 (R 2 = 0.9952) and with a detection limit of 0.54 μg ml −1 . d -PA content in human urine samples was detected with recoveries of 104.8–112.99%. Fluorescence-enhanced determination of d -PA using Cu NCs was established for the first time and this rapid, easy and sensitive method should attract much attention for this application.
研究领域:
分析化学
·
纳米技术
·
生物医学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES
·
2017
·
IF 0.697
Yanni Miao,Qi Liu,Wei Wang,Li Liu,Li Wang
A new chiral capillary electrophoresis (CE) analytical method for the determination of six pairs of amino acid enantiomer-derivatized precapillary with 9-fluorenylmethoxycarbonyl chloride (FMOC) was developed. CE separation parameters such as the pH of background electrolyte, the boric acid concentration, the addition of D-fructose and isopropanol, and the effect of new binary chiral selectors on the electropherograms were investigated. Precapillary and in-capillary derivatization was compared as well. The results showed that enantiomeric separations were obtained with enantio-resolution (Rs) ranged from 2.61 to 9.99 for the FMOC-derivatized amino acid enantiomers except alanine with Rs 1.06. Precapillary derivatization method provides overall better Rs for nearly all the targets except FMOC-aspartate with efficiency up to 1.309 × 106 plates and limit of detection (LOD) down to 2.5 μM. This method was successfully applied to analyze the concentration of the D/L-glutamic acid and D/L-aspartate in seven rice wine samples.
研究领域:
分析化学
·
食品科学
✓ 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）