BIOPROCESS AND BIOSYSTEMS ENGINEERING
·
2022
·Vol. 45
·期 6
·1075-1088
·
IF 3.434
Wei Kaixin,Wu Xiaomei,Ma Baodi,Li Zhi,Xu Yi
A series of nickel-incorporated SBA-15 mesoporous molecular sieves (Ni-SBA-15) were prepared as support for the immobilization of his-tagged recombinant Microbacterium esterase. The Ni-SBA-15 could strongly and specific absorb the his-tagged esterase from cell disrupted supernatant. It was found that the nickel amount in Ni-SBA-15 has dramatic influence on the activity and thermo-stability of immobilized enzyme, while the kinds of nickel precursor had little effect on enzyme stability. The morphology, chemical composition and structure of the best support NiCl 2 -SBA-15 (Ni-SBA-15 prepared from NiCl 2 precursor) were characterized by various spectroscopy techniques. The immobilized esterase retained full activity of free esterase and showed high immobilized yield (> 90%) with higher thermo-stability, pH stability and organic solvent resistance compared with free enzyme. The optimum reaction temperature increased from 35 to 40 °C and the optimal reaction pH moved from 10.0 to 8.0 after enzyme immobilization. The immobilized esterase exhibited excellent storage stability and keeping 92% of the initial activity after 30 days’ storage at 25 °C. In addition, the immobilized esterase had excellent reusability for the synthesis of key chiral intermediate of d -biotin and the substrate conversion could still keep 100% after 13 cycles continuously. Finally, optical pure (4 S , 5 R )-hemiester was obtained in 80.8% isolated yield and 99% purity in the gram preparative scale.
研究领域:
生物化学
·
酶工程
·
有机合成
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BIOPROCESS AND BIOSYSTEMS ENGINEERING
·
2021
·Vol. 44
·期 10
·2193-2204
·
IF 3.21
He Song,Wu Xiaomei,Ma Baodi,Xu Yi
The novel Ni-NTA-functionalized magnetic chitosan microspheres (MCS-NTA-Ni) were prepared via amino functionalization of MCS with epichlorohydrin and ethylenediamine, followed by the introduction of the aldehyde groups and NTA in turn, and nickel (II) ions were chelated in the end. MCS-NTA-Ni contained numerous long-armed NTA-Ni surface groups, ensuring high enzyme loading and providing more space and flexibility to attach enzymes and maintain their activity. This microsphere can have highly selective adsorption of his-tagged recombinant protein. The his-tagged recombinant Microbacterium esterase of E. coli BL21 (DE3)/pET21a-EstSIT01 was first immobilized on MCS-NTA-Ni by affinity fixation, giving high immobilization yield (90.1%) and enzyme loading (120 mg/g). Compared with free esterase, the immobilized esterase was found to exhibit higher pH stability and thermal stability. In addition, the immobilized esterase had excellent reusability for the synthesis of key chiral intermediate of d -biotin and the substrate conversion could still keep 100% after 8 cycles continuously.
研究领域:
生物化学
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酶工程
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合成生物学
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JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC
·
2013
·
IF 2.823
Jian-Yong Zheng,Sheng-Fan Wang,Yin-Jun Zhang,Xiang-Xian Ying,Yu-guang Wang,Zhao Wang
A chemoenzymatic methodology for the asymmetric synthesis of d -biotin intermediate lactone ((3a S , 6a R )-tetrahydro-1,3-dibenzylhexahydro-1 H -Furo[3,4-d] imidazole-2,4-dione) 1 has been demonstrated. The key step of the synthetic routes is Lipozyme RM IM catalyzed desymmetrization of meso-diols 3 . The highest enantiomeric excess ( e.e. > 98%) and yield (>90%) of the product was achieved with Lipozyme RM IM in Dioxane/Toluene (1:3, v/v) at 35 °C. Furthermore, Lipozyme RM IM showed an excellent operational stability, retaining above 80% of the initial activity after 10 cycles of reaction. d -Biotin intermediate lactone 1 was obtained subsequently by Jones oxidation, basic hydrolysis and lactonization.
研究领域:
生物化学
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有机合成
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酶工程
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Biomedical Optics Express
·
2024
·Vol. 15
·期 10
·5947-5959
·
IF 2.9
Mengjing Xu,Mengjing Xu,Boyang Shi,Haofeng Li,Xiaohan Mai,Lan Mi,Jiong Ma,Xiangdong Zhu,Guowei Wang,Guowei Wang,Yiyan Fei,Yiyan Fei,Yiyan Fei
Label-free optical biosensors have become powerful tools in the study of biomolecular interactions without the need for labels. High throughput and low detection limit are desirable for rapid and accurate biomolecule detection. The oblique-incidence reflectivity difference (OI-RD) technique is capable of detecting thousands of biomolecular interactions in a high-throughput mode, specifically for biomolecules larger than 1000 Da. In order to enhance the detection capability of OI-RD for small molecules (typically < 500 Da), we have developed a three-dimensional biochip that utilized carboxymethyl chitosan (CMCS) functionalized slides. By investigating various factors such as sonication time, protein immobilization time, CMCS molecular weight, and glutaraldehyde (GA) functionalization time, we have achieved a detection limit of 6.8 pM for avidin (68 kDa). Furthermore, accurate detection of D-biotin with a molecular weight of 244 Da has also been achieved. This paper presents an effective solution for achieving both high throughput and low detection limits using the OI-RD technique in the field of biomolecular interaction detection.
研究领域:
生物传感
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光学传感
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分析化学
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生物技术
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SENSORS AND ACTUATORS B-CHEMICAL
·
2023
·
IF 8.4
Shiqi Hu,Weicheng Shi,Yu Chen,Jinming Yuan,Xin Xiong,Tingting Liu,Shilei Ding,Wei Xiao,Yaofei Chen,Gui-Shi Liu,Zhe Chen,Lei Chen,Yunhan Luo
Surface plasmon resonance (SPR), generally including propagating SPR (PSPR) and localized SPR (LSPR), has attracted wide attention due to the uniqueness of the strong evanescent field. The PSPR has the merit of high sensitivity over the LSPR, but lacks flexibility to tune resonant wavelength and serves limited application. Here we revisit the fundamental PSPR wavevector matching theory and construct a three-dimensional (3D) tuning hypersurface for the parameters of resonant wavelength, incident angle, and surrounding refractive index , which provides the performance tuning guide map. With assistance of the hypersurface, a hypersurface-tuning PSPR (HT-PSPR) sensor is designed with high sensitivity and ultra-wide detection range. Experiments demonstrate that the sensor can selectively tune the resonance and render the sensitivity to a theoretical upper limit of ∼10 5 nm/RIU, or an ultra-wide detection range of 0.518 RIU, which satisfies the requirement for most gas and liquid sensing applications. Using D -biotin as the detection sample of small molecule (MW=244 Da), the HT-PSPR sensor achieves an ultra-low detection limit down to 27.5 fM. The hypersurface-tuning has created an intrinsic, universal and flexible method for tuning PSPR at the desired resonant wavelength, sensitivity and detection range, which is expected to find great significance in SPR design and applications.
研究领域:
光学
·
传感器
·
纳米技术
·
生物医学工程
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PHYTOMEDICINE
·
2025
·
IF 8.3
Zhi-Hua Deng,Liang-Jun He,Shanshan Wei,Chang-Sheng Li,Ya Chen,Xin-Yi Ai,Chatchai Chaotham,Supat Jiranusornkul,Pei-Cheng Zhang,Zhuo Luo,Shang-Ping Xing,Jie Yang
Background Targeting tumor metabolic reprogramming, particularly de novo lipogenesis (DNL) regulated by acetyl-CoA carboxylase 1 (ACC1), constitutes a promising therapeutic strategy. Non-small cell lung cancer (NSCLC) is a predominant subtype of lung cancer, characterized by limited treatment options and poor prognosis. Plumbagin (PLB), a natural naphthoquinone, exhibits broad antitumor activity. However, its direct targets and mechanisms in NSCLC remain unclear. Purpose This study aims to identify the molecular targets of PLB and elucidate its antitumor mechanism in NSCLC. Methods High-resolution mass spectrometry, in conjunction with a biotin-labeled PLB probe, was utilized to identify the direct targets of PLB. Surface plasmon resonance (SPR), streptavidin-based immunoprecipitation, and molecular dynamics simulation were employed to confirm the direct interaction between PLB and ACC1. ACC1 knockdown NSCLC cells were generated using siRNAs, followed by colony formation, wound-healing, and transwell assays. Tissue microarray analysis of clinical NSCLC samples was conducted to assess ACC1 expression and its clinical relevance. Lipid metabolites in NSCLC cells and tissues were quantified using an ELISA, and lipid droplet formation was observed via BODIPY staining. The in vivo therapeutic efficacy of PLB was evaluated using NSCLC xenograft models. Results Proteomic analysis identified ACC1 as a direct PLB target. Binding assays and SPR analyses confirmed the specific interaction between PLB and ACC1, and structural simulations revealed that PLB binds to the carboxyltransferase domain of ACC1 with high affinity. Importantly, tissue microarray analysis revealed that ACC1 was significantly overexpressed in tumor tissues of patients with NSCLC, and high expression of ACC1 was positively correlated with poor prognosis. Mechanistically, PLB inhibited the ACC1/FASN/SCD1 lipogenic axis, resulting in a substantial reduction in lipid metabolites and lipid droplet accumulation. Furthermore, exogenous palmitic acid supplementation partially reversed the antitumor effects of PLB, whereas ACC1 knockdown synergistically enhanced PLB’s antitumor efficacy. In NSCLC xenograft models, PLB significantly inhibited tumor growth by targeting the ACC1/FASN/SCD1 pathway and reducing key lipid metabolites. Conclusion Our findings identify PLB as a novel ACC1 inhibitor that suppresses NSCLC by targeting the ACC1/FASN/SCD1 axis-mediated DNL pathway, highlighting its potential as a metabolic modulator for NSCLC therapy.
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International Journal of Nanomedicine
·
2025
·
IF 6.5
Weiwei Ma,Yan Zhang,Xuejing Zhai,Qian Qu,Xueying Guo,Sen Zhang,Ruiyao Hou,Ping Lu,Yanyan Yin
Background Graphene oxide (GO) has high drug-loading capacity and good photothermal property. However, the limited stability and poor biocompatibility of GO hindered its application as drug delivery carrier for future nanomedicine.Methods In this study, a new strategy of using chemical conjugation on GO with polypeptide was adopted. A novel Biotin grafted polysarcosine polymers (B-PSar) modified graphene oxide derivative (B-PSar-GO) was successfully synthesized and utilized as a carrier to develop a new drug delivery system for targeted chemo-photothermal cancer therapy. In vitro and in vivo experiments evaluated the system’s biosafety and antitumor efficacy.Results With the B-PSar protection, the B-PSar-GO showed excellent biological safety with the average size of 268.2±8.4 nm. Stability experiments displayed B-PSar-GO was extremely stable. The anti-cancer drug doxorubicin (DOX) was loaded on B-PSar-GO through π–π interactions and hydrophobic interactions, B-PSar-GO@DOX achieved a maximum loading capacity of 25.5%. In addition, B-PSar-GO@DOX exhibited NIR/pH dual-responsive DOX release characteristics, ensuring sustained drug release to tumor tissues triggered by NIR laser irradiation and acidic tumor microenvironment. Based on the excellent photothermal conversion efficiency of GO, B-PSar-GO@DOX showed excellent chemo-photothermal synergistic tumor inhibition both in vitro and in vivo under NIR irradiation.Conclusion The novel nano-drug delivery system B-PSar-GO@DOX developed in this paper offers a promising platform for chemo-photothermal synergistic cancer treatment.
研究领域:
纳米医学
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癌症治疗
·
药物递送
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生物材料
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光热治疗
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MICROCHIMICA ACTA
·
2025
·Vol. 193
·期 1
·28
·
IF 5.3
Li Yixian,Yang Xiaohui,Cheng Shanshan,Mu Rongrong,Li Huiqiang,Yu Yang
Autoantibodies play a critical role in predicting diseases and monitoring therapeutic outcomes. Limitations of current autoantibody detection techniques include prolonged detection times and operational complexity. This study aims to develop a bidirectional microfluidic platform for the detection of autoantibodies. A “double inlet” and “waste outlet” structure was developed on the bidirectional lateral-flow microfluidic chip (BiLFMC). Using anti-citrullinated peptide antibodies (ACPA) as a model, this study validated the feasibility and performance advantages of the BiLFMC in the diagnosis of autoimmune disease. The BiLFMC was optimized by selecting the optimal distances between the sample inlet and waste outlet, as well as the ideal diameter for the waste outlet. The most suitable absorbent material was chosen for the waste absorbent pad. The detection and anti-interference performance of the unidirectional mode, unidirectional sequential mode, and bidirectional mode were compared, demonstrating that the bidirectional microfluidic chip significantly enhanced anti-interference capabilities. For ACPA detection, a serum sample required only a 1:100 dilution and 10 μL volume, with results available in 8 min. The coefficient of variation (CV) for repeatability ranged from 7.04% to 13.55%, while the CV for intermediate precision ranged from 11.06% to 14.49%. The limit of quantitation was 3.951 RU/mL, and the upper limit of the linear range was 500 RU/mL. When compared to ELISA results, the consistency coefficient was 0.8302. Furthermore, the BiLFMC was able to accurately distinguish samples exceeding the ELISA upper limit (> 196 RU/mL). A bidirectional microfluidic chip has been developed, incorporating indirect detection methods to enhance the anti-interference capability of the microfluidic platform. Graphical abstract
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TRENDS IN BIOTECHNOLOGY
·
2026
·
IF 16.6
Dianjiang Yu,Haofeng Chen,Yang Zhang,Yueyang Chen,Hongwei Chen,Shujin Lin,Junyi Wang,Haoyi Yang,Xueping Ling,Ning He,Jifeng Yuan
Microbial synthesis of chemicals still faces low production due to the complicated metabolic networks, which generally cannot be tackled by rational engineering strategies. In this study, we report on a workflow, biosensor-guided indexing of genome evolution (BRIDGE), to obtain marker-free engineered strains with improved chemical production. We adopt the BRIDGE system to achieve high-level protocatechuic acid (PCA) production in Pseudomonas putida , reaching 4.67 g l −1 in shake flasks. Transcriptomics analysis reveals a synergy between metabolic pathway activation and the secretion system, allowing PCA overproduction. Furthermore, we demonstrate that the engineered P. putida can serve as a platform for de novo synthesis of other chemicals with high titers, achieving 2.97 g l −1 cis,cis -muconate in shake flasks. Since the BRIDGE system gives marker-free engineered strains, it would greatly favor downstream industrial applications. Considering the great variety of available biosensors, we envision that our BRIDGE approach will accelerate other microbial synthesis processes with enhanced productivity.
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PHYTOMEDICINE
·
2026
·
IF 11.3
Qiang-Ming Li,Man-Lei Zhu,Xiang Li,Jing-Xing He,Xue-Qiang Zha,Jian-Ping Luo,Feng-Yun Zhang
Background Identifying novel targets that ameliorating dopaminergic neuron iron metabolism dysregulation may provide potential therapeutic strategies for Parkinson’s disease (PD). Buddlejasaponin IVb (BJP-IVb) is the first natural product that can ameliorate iron regulatory protein 2 (IRP2)-mediated iron metabolism dysregulation in dopaminergic neuron to suppress PD. BJP-IVb may exert this bioactivity through novel target. However, the pharmacological target of BJP-IVb is unclear. Purpose This study aimed to discovery the potential novel target of BJP-IVb ameliorating dopaminergic neuron iron metabolism dysregulation in PD. Method BJP-IVb probe (BJP-IVb-P) were synthesized to identify the potential novel target by affinity-based protein profiling. The expression of DEAD-box helicase 17 (DDX17) was knocked down in vitro and in vivo to prove the potential novel target of BJP-IVb is DDX17. Results We proved that BJP-IVb could directly bind to DDX17. Dopaminergic neurons exhibit an elevated expression of DDX17 in PD. Dopaminergic neuron iron metabolism dysregulation and Janus kinase 2 (JAK2) and signal transducer and activator of transcription 5 (STAT5) signaling pathway activation in PD could be alleviated by the knockdown of DDX17 in vitro and in vivo . The inhibitory effect of BJP-IVb on dopaminergic neuron iron metabolism dysregulation and JAK2–STAT5 signaling pathway activation in PD could be abolished by the knockdown of DDX17 in vitro and in vivo . Conclusion This study first proved that DDX17 is a pharmacological target for ameliorating dopaminergic neuron iron metabolism dysregulation in PD, and BJP-IVb ameliorates dopaminergic neuron iron metabolism dysregulation by targeting DDX17.
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2026
·
IF 6.2
Yan Wang,Yitian Yu,Nuohan Xu,Zhenyan Zhang,Bingfeng Chen,Minglong Song,Qi Zhang,Tingzhang Wang,Yunting Ma,Tao Lu,Liwei Sun,Haifeng Qian
B vitamins (VBs) are essential micronutrients, yet their microbial production across natural habitats remains poorly understood. We analyzed 126,931 metagenome-assembled genomes to map the distribution of VB producers and constructed a database of 87 VB synthesis genes, identifying VB-independent producers and their environmental drivers. Soil and freshwater habitats harbored the greatest diversity of VB1, VB2, VB7, VB9, and VB12 producers, whereas marine systems contained fewer. Biosynthetic capacity was rare for VB5 and was limited for VB3 and VB6. Gammaproteobacteria are prolific producers, synthesizing up to eight VB types. Soil sulfur content and conductivity significantly enhanced VB production potential, linking beneficial microbes to improved soil health. We further validated VB3 and VB7 synthesis in Acinetobacter kookii, supporting our predictive framework. Because the identified producers include both beneficial and pathogenic taxa, careful evaluation is required. This work provides a robust gene database and a foundation for optimizing microbial VB production in agriculture and biotechnology.
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ANALYTICAL CHEMISTRY
·
2026
·
IF 7.3
Kao Shangbin,Tian Nannan,Zhang Yaping,Qin Yuan,Gu Bing,Fang Hongming,Zhao Xueqin,Huang Biao
Small extracellular vesicles (sEVs) have emerged as promising
biomarkers
for liquid biopsy; however, the limited specificity of single-marker
detection and the intrinsic heterogeneity of circulating sEVs hinder
their reliable clinical application. Herein, we develop a programmable
dual-aptamer amplified luminescent proximity homogeneous assay (AlphaLISA)
for rapid and wash-free detection of tumor-derived sEVs. In this strategy,
EpCAM and PD-L1 aptamers simultaneously recognize two membrane proteins
on the same vesicle, enabling proximity-induced interaction between
donor and acceptor beads to generate a chemiluminescent signal via
singlet oxygen-mediated energy transfer. This dual-recognition design
effectively improves analytical specificity by implementing a molecular
“AND” logic requirement for signal generation. The proposed
assay exhibits a broad linear range of 5.3 × 10 5 to
5.3 × 10 9 particles mL –1 with a
limit of detection of 1.63 × 10 4 particles mL –1 . Good analytical performance was demonstrated by
recovery rates of 93.9–107.5% with relative standard deviations
below 5%, indicating high accuracy and reproducibility. Notably, the
assay enables signal readout within 1 min under homogeneous conditions,
highlighting its operational simplicity and rapid response. Clinical
applicability was evaluated using serum samples from 20 lung cancer
patients and 20 healthy controls, achieving an area under the ROC
curve of 0.87 (95% CI: 0.75–0.97), with a sensitivity of 95%
and a specificity of 75%. Overall, this programmable dual-aptamer
AlphaLISA platform provides a rapid, homogeneous, and highly adaptable
strategy for tumor-derived sEVs analysis, offering promising potential
for noninvasive cancer diagnostics
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