Biotechnology Journal
·
2024
·Vol. 19
·期 6
·2400040
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IF 3.2
Shibo Jiang,Ying Lin,Suiping Zheng
IMP (inosinic acid) is a crucial intermediate in the purine metabolic pathway and is continuously synthesized in all cells. Besides its role as a precursor for DNA and RNA, IMP also plays a critical or essential role in cell growth, energy storage, conversion, and metabolism. In our study, we utilized the circularly permuted fluorescent protein (cpFP) and IMP dehydrogenase to screen and develop the IMP biosensor, IMPCP1. By introducing a mutation in the catalytically active site of IMPCP1, from Cys to Ala, we disrupted its ability to catalyze IMP while retaining its capability to bind to IMP without affecting the IMP concentration in the sample. To immobilize IMPCP1, we employed the SpyCatcher/SpyTag system and securely attached it to Magarose-Epoxy, resulting in the development of the IMP rapid test kit, referred to as IMPTK. The biosensor integrated into IMPTK offers enhanced stability, resistance to degradation activity, and specific recognition of IMP. It is also resistant to peroxides and temperature changes. IMPTK serves as a rapid and stable assay for analyzing IMP concentrations in fermentation broth. Within the linear range of IMP concentrations, it can be utilized as a substitute for HPLC. The IMPTK biosensor provides a reliable and efficient alternative for monitoring IMP levels, offering advantages such as speed, stability, and resistance to environmental factors.
研究领域:
生物化学
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生物技术
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分子生物学
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DOI
PubMed
高影响因子
Food Bioscience
·
2021
·
IF 4.24
Jiatong Li,Genghui Zhou,Peng Xue,Xiuping Dong,Yongtao Xia,Joe Regenstein,Ming Du,Liming Sun
Bacterial spoilage is an important reason causing freshness deterioration of fish during postmortem storage. The aim of this study is mainly to investigate the role of bacteria in freshness and the degradation of inosine 5′-monophosphate (IMP) as well as nucleotide derived values in chilled sturgeon (hybrid, Acipenser schrenckiid × Huso dauricus ) fillets using a microbial reduction pretreatment. During storage at 4 or -1 °C, lower total bacterial count and total volatile base nitrogen in the treated samples was observed when compared to the control samples. But very close K, Ki value and IMP was observed in both groups, although higher IMP was recorded at day 6 and 9 at 4 °C. In later storage period, accelerated decrease of inosine (Ino) and increment of hypoxanthine (Hx) occurred in two control groups, resulting in a rapid increase of G and H values, significant higher than the treated groups. Six predominant spoilage bacteria ( Acinetobacter johnsonii , Acinetobacter lwoffii , Shewanella baltica , Pseudomonas Fragi , Pseudomonas koreensis and Pseudomonas antarctica ) were isolated from the sturgeon fillets during storage. Based on the results of respective co-cultivation of IMP with the six species, only Acinetobacter johnsonii had ability to degrade IMP when reached a high density of 2.84 × 10 9 CFU/mL. Results suggested that microbial reduction pretreatment could better the freshness of chilled sturgeon fillet. Comparing to endogenous IMPase, spoilage bacteria probably played a small role in IMP degradation, but affected considerably the decomposition of Ino to Hx.
研究领域:
食品科学
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微生物学
·
水产养殖
·
渔业
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DOI
高影响因子
Chemical Biology & Drug Design
·
2021
·Vol. 99
·期 2
·362-372
·
IF 2.817
Jia-Zhu Chigan,Jia-Qi Li,Huan-Huan Ding,Yin-Sui Xu,Lu Liu,Cheng Chen,Ke-Wu Yang
Bacterial resistance caused by metallo-β-lactamases (MβLs) has become an emerging public health threat, and the development of MβLs inhibitor is an effective way to overcome the resistance. In this study, thirteen novel O -aryloxycarbonyl hydroxamates were constructed and assayed against MβLs. The obtained molecules specifically inhibited imipenemase-1 (IMP-1) and New Delhi metallo-β-lactamase-1, exhibiting an IC 50 value in the range of 0.10–18.42 and 0.23–22.33 μM, respectively. The hydroxamate 5 was found to be the most potent inhibitor, with an IC 50 of 0.1 and 0.23 μM using meropenem and cefazolin as substrates. ICP-MS analysis showed that 5 did not coordinate to the Zn(II) ions at the active site of IMP-1, while the rapid dilution, thermal shift and MALDI-TOF assays revealed that the hydroxamate formed a covalent bond with the enzyme. Cytotoxicity assays indicated that the hydroxamates have low toxicity in MCF-7 cells. This work provided a potent scaffold for the development of MβLs inhibitors.
研究领域:
微生物学
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药物化学
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感染病学
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DOI
PubMed
高影响因子
CHEMICAL & PHARMACEUTICAL BULLETIN
·
2021
·Vol. 69
·期 12
·1179-1183
·
IF 1.645
Taichi Kamo,Keiichi Kuroda,Shota Kondo,Usaki Hayashi,Satoshi Fudo,Tomoki Yoneda,Akiko Takaya,Michiyoshi Nukaga,Tyuji Hoshino
Metallo-β-lactamases (MBLs) are significant threats to humans because they deteriorate many kinds of β-lactam antibiotics and are key enzymes responsible for multi-drug resistance of bacterial pathogens. As a result of in vitro screening, two compounds were identified as potent inhibitors of two kinds of MBLs: imipenemase (IMP-1) and New Delhi metallo-β-lactamase (NDM-1). The binding structure of one of the identified compounds was clarified by an X-ray crystal analysis in complex with IMP-1, in which two possible binding poses were observed. Molecular dynamics (MD) simulations were performed by building two calculation models from the respective binding poses. The compound was stably bound to the catalytic site during the simulation in one pose. The binding model between NDM-1 and the compound was constructed for MD simulation. Calculation results for NDM-1 were similar to those of IMP-1. The simulation suggested that the binding of the identified inhibitory compound was also durable in the catalytic site of NDM-1. The compound will be a sound basis for the development of the inhibitors for MBLs.
研究领域:
微生物学
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结构生物学
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药物化学
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DOI
PubMed
高影响因子
CELLULAR AND MOLECULAR LIFE SCIENCES
·
2022
·Vol. 79
·期 8
·1-15
·
IF 9.207
Chang Chia-Chun,Peng Min,Zhong Jiale,Zhang Ziheng,Keppeke Gerson Dierley,Sung Li-Ying,Liu Ji-Long
The cytoophidium is a unique type of membraneless compartment comprising of filamentous protein polymers. Inosine monophosphate dehydrogenase (IMPDH) catalyzes the rate-limiting step of de novo GTP biosynthesis and plays critical roles in active cell metabolism. However, the molecular regulation of cytoophidium formation is poorly understood. Here we show that human IMPDH2 polymers bundle up to form cytoophidium-like aggregates in vitro when macromolecular crowders are present. The self-association of IMPDH polymers is suggested to rely on electrostatic interactions. In cells, the increase of molecular crowding with hyperosmotic medium induces cytoophidia, while the decrease of that by the inhibition of RNA synthesis perturbs cytoophidium assembly. In addition to IMPDH, CTPS and PRPS cytoophidium could be also induced by hyperosmolality, suggesting a universal phenomenon of cytoophidium-forming proteins. Finally, our results indicate that the cytoophidium can prolong the half-life of IMPDH, which is proposed to be one of conserved functions of this subcellular compartment.
研究领域:
细胞生物学
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分子生物学
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生物化学
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DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2021
·
IF 7.514
Saili Zhang,Xiangjun Fang,Weijie Wu,Chuan Tong,Hangjun Chen,Hailong Yang,Haiyan Gao
Fresh shiitake ( Lentinus edodes ) is prone to brown, pileus-opening and flavor-loss during storage. Therefore, it is important to find an effective preservation method for fresh shiitake. Negative air ions (NAI) are negatively-charged molecules or atoms in the air, and can affect the physiological metabolism of live cells and be conveniently used with low cost. In this study, NAI treatment was performed at different times and the physico-chemical characteristics, microstructure, membrane potential and energy metabolism of shiitake were determined during storage. Results showed that NAI treatment for 40 min could reduce 29% of browning index and maintain the hardness of shiitake. NAI treatment groups had higher content of sweetness amino acids, umami amino acids, 5′-IMP, eight-carbon alcohols compounds and cyclic sulfides compounds than the control, and comprehensive quality of the group being treated for 40 min was the best. The mitochondria of shiitake swelled and the membrane potential decreased after being treated by NAI. However, NAI treatment for 40 min could improve the contents of ATP and ADP, maintain a relatively stable energy charge level, and promote energy utilization of shiitake during storage. The results demonstrated that NAI treatment had the potential to improve the quality shiitake during storage.
研究领域:
食品科学
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微生物学
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植物生理学
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DOI
PubMed
高影响因子
NEW JOURNAL OF CHEMISTRY
·
2018
·Vol. 42
·期 24
·20180-20189
·
IF 3.201
Jinping Song,Qi Ma,Sufang Zhang,Huijun Liu,Yong Guo,Feng Feng
In this paper, we reported a facile approach for the preparation of sulfur and nitrogen co-doped carbon nanoparticles (S,N-CNPs) with strong blue luminescence, which exhibited multiple applications including metal ion detection, logic gate fabrication and bioimaging. S,N-CNPs were synthesized by one-step hydrothermal treatment of formamidine sulfinic acid (FAS) and ethylene glycol. The as-synthesized S,N-CNPs possess a high quantum yield (Φ = 69.04%) and display excellent photostability and storage stability. By modulating the pH environment, S,N-CNPs were successfully applied as fluorescent probes for the detection of three kinds of heavy metal ions (Hg2+, Ag+ and Ni2+). The fabricated fluorescence sensors display ultrahigh sensitivity and a wider signal response range. More importantly, ethylenediamine (or ammonium thioglycolate and glycine) could efficiently release Hg2+ (or Ag+ and Ni2+) from the S,N-CNP surface, leading to the recovery of the fluorescence. According to this phenomenon, interesting implication (IMP) logic gates were constructed. In addition, the S,N-CNPs exhibited low toxicity, good biocompatibility and easy cellular uptake behavior. Consequently, the obtained S,N-CNPs will have potential applications in multiple fields of environmental monitoring, logic gate operation, and cell bioimaging.
研究领域:
材料科学
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分析化学
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纳米技术
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环境监测
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生物医学工程
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DOI
高影响因子
ACS Applied Materials & Interfaces
·
2025
·
IF 8.2
Li Liao,Yufei Liu,Xianhai Li,Zewei Jiang,Zhijie Jiang,Jing Yao
Effective tumor immunotherapy is hindered by an immunosuppressive tumor microenvironment (TME), especially in triple-negative breast cancer. Though phototherapy could induce immunogenic cell death (ICD) to increase antitumor immunity, the simultaneous upregulation of indoleamine 2,3-dioxygenase (IDO) induces the negative immunomodulatory effect termed as the “immune-metabolism” loop to compromise immunotherapeutic efficacy. Herein, we developed IMMGP consisting of biomimetic IND-Mn@PM (IDP) and ICG-MnO2@PM (IMP), which combines the phototherapy-induced ICD and metabolic reprogramming to solve the dilemma. During the light-on phase, IMP effectively kills cancer cells with potent photodynamic ROS generation with the assistance of MnO2-produced oxygen and induces ICD to reverse the immunosuppressive TME. In the light-off phase, Mn2+ (from IDP and MnO2-based redox reaction) elicits a Fenton-like reaction to relay ROS generation, which is further orchestrated with continuous exhaustion of intratumoral GSH by the conversion of Mn3+ to Mn2+, and promotes dendritic cell maturation. Moreover, the released indoximod (IND) downregulated IDO to inhibit kynurenine metabolism, which reinvigorates T cell-mediated antitumor immunity. Collectively, IMMGP amplifies the immune response by breaking the “immune-metabolism” loop and sustaining the “immunologically hot” state after phototherapy, thus leading to nearly complete tumor inhibition (94.25%). Thus, IMMGP-mediated dual-phase photodynamic immunotherapy offers a novel approach in cancer nanomedicine.
研究领域:
纳米医学
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肿瘤免疫学
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生物材料学
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产品关联
DOI
PubMed
高影响因子
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