MICROCHEMICAL JOURNAL
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2025
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IF 5.1
Ting Zhang,Danni Ma,Fuman Cai
The accurate detection of glucose is critical for clinical diagnostics and metabolic monitoring. This study focused on the successful synthesis and incorporation of the glucose dehydrogenase (GDH) gene sequence from Burkholderia cepacia into a plasmid, followed by codon optimization and modification of cysteine motifs for increased expression in E. coli . The engineered E. coli strain demonstrated efficient heterologous expression of stable GDH. Remarkably, the recombinant strain not only enabled cost-effective GDH production but also exhibited excellent glucose catalytic activity, with a detection range of 1 μM to 10 mM and a minimum detectable concentration of 1 μM. This functionality makes it suitable for applications of AuNPs/SPE/glucose biosensors, offering high sensitivity for glucose detection—a novel achievement not previously reported. These findings have the potential to advance the clinical utilization of AuNPs/tetracysteine/GDH-modified biosensors and may promote the development of GDH-based electrochemical biosensors.
研究领域:
生物化学
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生物技术
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分析化学
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分子生物学
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纳米技术
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临床诊断
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代谢监测
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产品关联
DOI
高影响因子
ANALYTICAL BIOCHEMISTRY
·
2016
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IF 2.243
Wei Zhang,Min Zhu,Feng Wang,Danhui Cao,Jennifer Jin Ruan,Weike Su,Benfang Helen Ruan
Glutamate dehydrogenase (GDH) catalyzes the oxidative deamination of L-glutamate and is important for several biological processes . For GDH inhibitor screening, we developed a novel mono-sulfonated tetrazolium salt (EZMTT), which can be synthesized using H 2 O 2 oxidation and purified easily on silica gel in large quantities. The EZMTT detection method showed linear dose responses to NAD(P)H, dehydrogenase concentration and cell numbers. In E. coli GDH assay, the EZMTT method showed excellent assay reproducibility with a Z factor of 0.9 and caused no false positives in the presence of antioxidants (such as BME). Using the EZMTT-formazan-NAD(P)H system, we showed that EGCG is a potent E. coli GDH inhibitor (IC 50 45 nM) and identified that Ebselen , a multifunctional thioredoxin reductase inhibitor, inactivated E. coli GDH (IC 50 213 nM). In cell-based assays at 0.5 mM tetrazolium concentration, EZMTT showed essentially no toxicity after a 3-day incubation, whereas 40% of inhibition was observed for WST-8. In conclusion, EZMTT is a novel tetrazolium salt which provides improved features that are suitable for dehydrogenases and real-time cell-based high-throughput screening (HTS).
研究领域:
生物化学
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药物筛选
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细胞生物学
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产品关联
DOI
PubMed
高影响因子
Biology-Basel
·
2026
·Vol. 15
·期 18
·1652
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IF 4.3
Haoyu Chai,Jutao Li,Cuiluan Ma,Yu-Cai He
A sequential two-stage chemo-biocatalytic process was investigated for the conversion ofD-xylose to furfuryl alcohol (FOL), an important furan derivative widely used in the manufacture of resins, polymers, fuels, and other value-added products. In the first stage,D-xylose was dehydrated to furfural (FAL) in a ternary Betaine:Formic acid:Malonic acid–water (Bet:FA:MA–H2O) reaction medium. Under the selected conditions, a FAL yield of 65.8% was obtained from 22.5 g/LD-xylose using 15 wt% Bet:FA:MA at 170 °C for 30 min. For the subsequent bioreduction stage, a recombinantEscherichia colistrain co-expressing the aldehyde reductase EutG and glucose dehydrogenase (GDH) was constructed. Using glucose as a co-substrate, EutG–GDH whole cells were able to reduce commercial FAL at concentrations up to 150 mM under the selected conditions of 40 °C and pH 7.5. For the sequential process using theD-xylose-derived dehydration liquor, the acidic reaction mixture was adjusted to pH 7.5 and diluted from 98.7 to 30.0 mM FAL before whole-cell bioreduction. The conditioned 30.0 mM FAL feed was essentially completely converted to FOL within 24 h. These results demonstrate the laboratory-scale feasibility of sequentially combining Bet:FA:MA–H2O-mediatedD-xylose dehydration with EutG–GDH whole-cell FAL reduction after interstage pH adjustment and dilution.
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DOI
高影响因子
ELECTROCHIMICA ACTA
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2016
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IF 4.803
Chuantao Hou,Qiaolin Lang,Aihua Liu
Redox polymer hydrogels have been proved to be very promising for biofuel cells design and electrochemical biosensing. Here we report that a facile hydrogel bioanode based on electron-withdrawing nitro-group tailored 1,4-naphthoquinone (1,4-NQ)-appended redox polymer and flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH) can dramatically improve the catalytic performance of glucose oxidation , for which a high current density of 1.97 ± 0.06 mA cm −2 and low onset potential of −0.13 V (vs. Ag/AgCl) can be achieved in 50 mM glucose solution without using any nanomaterials as electrode supports. Further, both electron-donating groups modified 1,4-NQs and electron-withdrawing nitro-group 1,4-NQs were synthesized to investigate their electrochemical properties. It was found that the enhanced catalytic performance can be mainly contributing from the nitro group-modified 1,4-NQs appended redox polymer, which shifted the formal potential in the positive direction and provided sufficient electrochemical driving force for glucose oxidation. Finally, this hydrogel bioanode was combined with a bilirubin oxidase based biocathode to construct a single-compartment glucose/O 2 biofuel cell with a high power output of 0.28 ± 0.03 mW cm −2 and open circuit potential of 0.69 V. Moreover, the O 2 -insensitive FAD-GDH could improve the cell stability obviously compared with glucose oxidase based cell. This study provides an alternative route to develop glucose biosensor or biofuel cell for directly harvesting energy from glucose.
研究领域:
生物化学
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电化学
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能源材料
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产品关联
DOI
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
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2022
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IF 3.705
Liya Zhou,Yaping Ouyang,Weixi Kong,Teng Ma,Hao Zhao,Yanjun Jiang,Jing Gao,Li Ma
In this study, a novel kind of Ni-NTA modified monodispersed SiO 2 nanoflowers (Ni-NTA@SiO 2 nanoflowers) were successfully synthesized. The obtained Ni-NTA@SiO 2 nanoflowers were used to specifically adsorb and purify His-tagged old yellow enzyme (OYE1) and glucose dehydrogenase (GDH), which allows access to optically pure (3 S)− 3-methyl-cyclohexanone through asymmetric hydrogenation reaction, and forms a cofactor regeneration system. The protein loading amount on Ni-NTA@SiO 2 nanoflowers was 40.17 mg/g support and the activity recoveries of OYE1 and GDH were 81.53% and 79.68%, respectively. The effects of pH and temperature on the activity of free and co-immobilized enzymes were investigated, and the stability as well as reusability were also measured. Compared to free enzymes, the co-immobilized enzymes showed higher thermal and storage stability. The co-immobilized enzymes were applied to asymmetric reduction of C C bonds for the synthesis of a chiral center with excellent enantioselectivity (ee > 99%), and the conversion was 46.02% after 7 cycles. This work introduced a one-pot multi-enzyme purification and co-immobilization strategy to construct efficient cofactor regeneration system with high activity and stability.
研究领域:
生物化学
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酶工程
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纳米材料科学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2017
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IF 6.32
Min Zhu,Jinzhang Fang,Jingjing Zhang,Zheng Zhang,Jianhui Xie,Yan Yu,Jennifer Jin Ruan,Zhao Chen,Wei Hou,Gensheng Yang,Weike Su,Benfang Helen Ruan
Glutaminase (KGA/isoenzyme GAC) is an emerging and important drug target for cancer. Traditional methods for assaying glutaminase activity are coupled with several other enzymes. Such coupled assays do not permit the direct and stringent characterization of specific glutaminase inhibitors. Ebselen was identified as a potent 9 nM KGA inhibitor in the KGA/glutamate oxidase (GO)/horse radish peroxidase (HRP) coupled assay but showed very weak activity in inhibiting the growth of glutamine-dependent cancer cells. For rigorous characterization, we developed a direct kinetic binding assay for KGA using bio-layer interferometry (BLI) as the detection method; Ebselen was identified as a GDH inhibitor but not a KGA inhibitor. Furthermore, we designed and synthesized several benzo[d][1,2]selenazol-3(2H)-one dimers which were subjected to SAR analysis by several glutaminolysis specific biochemical and cell based assays. Novel glutamate dehydrogenase (GDH) or dual KGA/GDH inhibitors were discovered from the synthetic compounds; the dual inhibitors completely disrupt mitochondrial function and demonstrate potent anticancer activity with a minimum level of toxicity.
研究领域:
癌症研究
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生物化学
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分子生物学
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药物发现
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DOI
PubMed
高影响因子
WASTE MANAGEMENT
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2023
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IF 8.1
Lin Zhu,Caihong Huang,Wei Li,Weixia Wu,Zhurui Tang,Yu Tian,Beidou Xi
Nitrogen loss from compost is a serious concern, causing severe environmental pollution. The NH 4 + -N content reflects the release of NH 3 . However, the nitrogen conversion pathway that has the greatest impact on NH 4 + -N content is still unclear. This study attempted to explore the key pathways, core functional microorganisms, and mechanisms involved in the transformation of ammonia nitrogen during composting. KEGG (Kyoto Encyclopedia of Genes and Genomes) metabolic pathways revealed that ammonia assimilation was dominated by the glutamate dehydrogenase (GDH) pathway (53.4%), which is crucial for nitrogen preservation. The combined analysis of KEGG, NR species annotation, and co-occurrence network identified 20 easy-to-regulate obligate core nitrogen-transforming functional microorganisms, including 18 ammonia-assimilating bacteria. Furthermore, the effects of environmental parameters on the obligate core functional microorganisms were investigated. The present study results provided a theoretical basis for the utilization of ten ammonia-assimilating bacteria, such as Paenibacillus , Erysipelatoclostridium, and Defluviimonas to improve the quality of compost.
研究领域:
环境科学
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微生物学
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堆肥技术
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DOI
PubMed
高影响因子
Biotechnology Journal
·
2024
·Vol. 19
·期 4
·2300557
·
IF 3.2
Han-Yu Liu,Feng Qian,Hai-Min Zhang,Qian Gui,Yao-Wu Wang,Pu Wang
The halogenase-based catalysis is one of the most environmentally friendly methods for the synthesis of halogenated products, among which flavin-dependent halogenases (FDHs) have attracted great interest as one of the most promising biocatalysts due to the remarkable site-selectivity and wide substrate range. However, the complexity of constructing the NAD + -NADH-FAD-FADH 2 bicoenzyme cycle system has affected the engineering applications of FDHs. In this work, a coenzyme self-sufficient tri-enzyme fusion was constructed and successfully applied to the continuous halogenation of L -tryptophan. Sp FDH was firstly identified derived from Streptomyces pratensis , a highly selective halogenase capable of generating 6-chloro-tryptophan from tryptophan. Then, using gene fusion technology, Sp FDH was fused with glucose dehydrogenase (GDH) and flavin reductase (FR) to form a tri-enzyme fusion, which increased the yield by 1.46-fold and making the coenzymes self-sufficient. For more efficient halogenation of L -tryptophan, a continuous halogenation bioprocess of L -tryptophan was developed by immobilizing the tri-enzyme fusion and attaching it to a continuous catalytic device, which resulted in a reaction yield of 97.6% after 12 h reaction. An FDH from S. pratensis was successfully applied in the halogenation and our study provides a concise strategy for the preparation of halogenated tryptophan mediated by multienzyme cascade catalysis.
研究领域:
生物技术
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生物化学
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酶学
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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）