INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Kai Yang,Yueshan Huang,Liyi Yao,Bin Huang,Weimin Zeng,Jin Huang
D-amino acid oxidase (DAAO) has long been recognized as a promising catalyst for chiral amino acid biosynthesis owing to its pronounced stereospecificity. However, its limited thermostability restricts industrial utility and poses challenges for protein engineering. Herein, we engineered a DAAO variant, M5 (S42D/T84D/H138N/A193R/E195Y/C234L), via a ProteinMPNN-guided multiple codon saturation mutagenesis approach. This variant exhibited superior catalytic efficiency (27.3 mM −1 s −1 , 29-fold relative to the starting enzyme) toward the non-natural compound D-glufosinate (D-PPT) and enhanced thermostability (519 min half-life at 50 °C, 208-fold relative to the starting enzyme). The evolution also expanded the enzyme's substrate scope toward several D-amino acids with different properties. The variant was proven able to completely transform 500 mM D-PPT in 7 h at a 100 mL scale with excellent stereospecificity. Molecular dynamics (MD) simulations gave the insight that the enhanced performance stems from altered surface charges and intramolecular/intermolecular networks. This strategy thus holds potential for engineering other enzymes involved in the biosynthesis of valuable products while reducing the screening costs during directed evolution.
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ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Huixiang Huo,Xuhua Wei,Chao Peng,Jing Li,Erkang Wang
Optimization of p-d orbital hybridization enhances charge interactions through increased orbital overlap, reduced electron transfer energy barriers, and regulated reaction activation energies, thereby significantly improving the catalytic kinetics. To implement the strategy, atomic substitution of Ru for Fe is employed in parent FeMnCuCoNi high-entropy nanozymes (HEzymes), yielding the RuMnCuCoNi HEzymes variant with outstanding peroxidase-like activity, demonstrating a 7-fold increase in activity over the parent alloy. Mechanistic studies reveal that the Ru 4dyz orbital, as the optimal catalytic site, exhibits stronger hybridization with the pz orbital of H2O2, which significantly enhances substrate adsorption. Concurrently, the reduced electron occupancy in antibonding orbitals strengthens orbital interactions with H2O2 while elevated electron density near the Fermi level accelerates electron transfer. The tripartite synergy, including enhanced adsorption, strengthened orbital interactions, and facilitated electron transfer, collectively drives the catalytic enhancement. Leveraging this nanozyme, an on-site detection platform is constructed for gastric cancer biomarkers (d-proline and d-alanine), achieving exceptional sensitivity and selectivity. Our work underscores the significance of p-d orbital hybridization regulation, providing a rational design principle for high-performance nanozymes.
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CURRENT PHARMACEUTICAL BIOTECHNOLOGY
·
2023
·Vol. 24
·期 14
·1803-1811
·
IF 2.8
Zou Min-Shu,Song Qiu-Ju,Yin Tai-Yong,Xu Hong-Tao,Nie Guo-Ming
Background: The etiology of systemic lupus erythematosus (SLE) is complex, and the disease is thus difficult to cure. In this regard, it has been established that SLE patients are characterized by differing levels of vitamin D-hydroxylation; however, the direct effects of vitamin D
(VitD) in these patients remain unknown. Objective: Therefore, we investigated the effects and mechanisms of action of VitD in the context of SLE. Methods: The effects of VitD on MRL/LPR mice were studied by synthesizing glycogen synthase kinase-3β (GSK-3β)-interfering lentiviruses
and transfecting with miR-126a-5p mimics. Changes in the body weight of mice were recorded for 6 weeks. Western blotting was performed to determine the levels of T-bet, GATA3, and GSK-3β protein expression, and qRT-PCR was performed to determine the levels of miR-126a-5p and GSK-3β
mRNA expression. ELISA was performed to determine the levels of ANA, dsDNA, and snRNP/Sm in mice serum. Results: GSK-3β and miR-126a-5p were expressed at high and low levels, respectively, in MRL/LPR mice. VitD (30 ng/kg) was found to reduce the expression of GSK-3β and increase
miR-126a-5p expression, which targets GSK-3β. T-bet and GATA3 were found to be positively regulated by miR-126a-5p and VitD and negatively regulated by GSK-3β. The body weight of mice was not altered by VitD. ANA, dsDNA, and snRNP/Sm were positively regulated by miR- 126a-5p and
VitD and negatively regulated by GSK-3β. The effects of GSK-3β were enhanced in response to the inhibition of miR-126a-5p expression. Conclusion: VitD upregulated miR-126a-5p to target GSK-3β expression, thereby alleviating the SLE in MRL/LPR mice.
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ENZYME AND MICROBIAL TECHNOLOGY
·
2020
·
IF 3.448
Jia-Qin Wu,Xiao-Mei Xu,Ding-Lin Wang,Neng-Bing Long,Rui-Feng Zhang
Novel nano-composites were prepared by coating epoxy resin-based cationic polymer in nano-thickness via in-situ curing on the nano-wall of macroporous SiO 2 with pore size of 0.5∼1 μm. By changing the thickness of polymer coating the specific surface area and porosity varied in range of 115∼74 m 2 /g and 90.4∼83.9 %, respectively. Through ion exchange phospholipase D (PLD, from Streptomyces sp ) was efficiently immobilized on the nano-composites as support and the immobilized PLD was applied for the highly efficient synthesis of phosphatidylserine (PS). The loading amount of PLD on the nano-composited support reached to a maximum of 90.2 mg/g support , 4 times as high as that on the pure macroporous silica. The specific activity of the immobilized PLD reached as high as 16,230 U/g protein , while that of free PLD was 18,780 U/g protein . Under a wide range of temperature and pH the stability and activity of the immobilized PLD were greatly improved as compared with the free ones. Under optimized conditions at 45 °C and pH 7.0, the PS yield reached as high as 96.2 % within 40 min. After 28 days storage the immobilized PLD retained 82.2 % of original activity, and after 12 cycling reuses it retained 79.3 % of PS yield, which indicated that the immobilized PLD exhibited good stability.
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Redox Biology
·
2019
·
IF 7.793
Shen-Fei Wang,Xin Liu,Mo-Yu Ding,Shuangcheng Ma,Jing Zhao,Ying Wang,Shaoping Li
Reducing agents are crucial for the management of maladaptive inflammation-induced macrophage death and hematopoietic toxicity of chemotherapy. 2- O - β - d -glucopyranosyl- l -ascorbic acid (AA-2 β G), a unique AA (or vitamin C) derivative identified in Lycium barbarum , exhibited enhanced free radical scavenging activity compared with AA and its synthetic derivative AA-2 α G. AA-2 β G protected hydrogen peroxide-induced cell death in murine macrophage RAW264.7 cells. Treatment with AA-2 β G eliminated oxidative stress and the ratio of cellular glutathione to glutathione disulfide more effectively than AA and AA-2 α G. AA-2 β G also significantly reduced the fluorescent intensity of DCFH-DA triggered by chemotherapeutic agent camptotehcin-11 but not fluorouracil. AA, AA-2 α G, and AA-2 β G significantly decreased Keap-1expression, and increased the expression levels of nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1. All compounds triggered the nuclear translocation of Nrf2, while the ability of AA-2 β G to enhance the Nrf2-DNA binding affinity was approximately two fold as those of AA and AA-2 α G. Sodium ascorbate cotransporters (SVCT) inhibitors, sulfinpyrazone, phloretin, and 3- O -methyglucose, potently abrogated the free radical scavenging activities of AA, AA-2 α G, and AA-2 β G. The cellular uptake efficacy of AA-2 α G and AA-2 β G was less than 10% of AA, while the inhibition of SVCT with sulfinpyrazone considerably diminished the uptake efficacy of these compounds. AA-2 α G and AA-2 β G are more stable in the Fenton reagents than AA. In summary, AA-2 β G from L. barbarum with excellent free radical scavenging activity is a promising natural AA derivative for further pharmacological evaluation.
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Molecular Medicine Reports
·
2019
·Vol. 20
·期 1
·332-340
·
IF 1.851
Lina Liu,Jing Yan,Fei Ge,Xiangtao Xu,Jun Lu,Huilian Shi,Shuihong Li,Yue Zhao,Changzheng Zhang
Saikosaponin‑D (SSD), which is the main bioactive component in the traditional Chinese medicine Chai Hu (Bupleurum falcatum L), possesses estrogen‑like properties and is widely used in treating estrogen‑related neurological disorders. The current study aimed to investigate the protective effects of SSD on the fear memory deficit in ovariectomized (OVX) rats and the potential underlying mechanism. SSD treatment significantly prolonged freezing time in OVX rats in a manner similar to that of estradiol (E2), whereas this effect was markedly suppressed by co‑administration of ICI182780, a non‑selective estrogen receptor (ER) inhibitor. The expression of ERα in the hippocampus of OVX rats was significantly elevated by SSD; however, Erβ expression and E2 synthesis were not markedly affected by SSD treatment. Collectively, this study demonstrated that SSD‑mediated fear memory improvement in OVX rats may be attributed not to E2 levels or ERβ activity, but to ERα activation in the hippocampus.
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PHARMACOLOGICAL RESEARCH
·
2020
·
IF 5.893
Xiaotian Qi,Mengyue Fan,Nana Huang,Xinyu Zhang,Jing Liu,Xiaoyu Li,Rong Sun
Cancer chemotherapy induced neutropenia (CCIN) is one of the most common toxicity caused by cytotoxic anticancer agents . Despite granulocyte colony-stimulating factor (GCSF) is widely used in clinical practice, the infection and infection-related mortality rate is still high for lack of functionally mature neutrophils. Saikosaponin d (SSD) is one of the major bioactive constituents of Radix Bupleuri (RB), which exerts immune-modulatory properties. We explored the function of SSD in CCIN therapy, we found that SSD contributed to generate functional mature neutrophils which capable of fighting infection both in vitro and in vivo . Network pharmacology was employed to explore the mechanism, 61 signal pathways might play an important role in CCIN treatment. Western Blot was employed to further confirm the potential pathway involved. We found CBL-ERK1/2 pathway was activated by SSD, followed by upregulating PU.1 and CEBPβ expression and leading to neutrophil differentiation. Our findings suggest a natural regimen SSD which could regenerate microbicidal neutrophils to effectively reduce CCIN-associated infection via activating CBL-ERK1/2, providing a rationale for future therapeutic approaches.
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CELLULAR AND MOLECULAR NEUROBIOLOGY
·
2016
·Vol. 37
·期 5
·869-877
·
IF 2.328
Shen Hui,Wang Jie,Jiang Dan,Xu Pei,Zhu Xiaolu,Zhang Yuanyuan,Yu Xing,Won Moo-Ho,Su Pei Qing,Yan Bing Chun
Some anticonvulsant drugs are associated with cognitive ability in patients; Topiramate (TPM) is well known as an effective anticonvulsant agent applied in clinical settings. However, the effect of TPM on the cognitive function is rarely studied. In this study, we aimed to observe the effects of TPM on cell proliferation and neuronal differentiation in the dentate gyrus (DG) of the d -galactose-induced aging mice by Ki-67 and doublecortin (DCX) immunohistochemistry. The study is divided into four groups including control, d -galactose-treated group, 25 and 50 mg/kg TPM-treated plus d -galactose-treated groups. We found, 50 mg/kg (not 25 mg/kg) TPM treatment significantly increased the numbers of Ki-67 + cells and DCX immunoreactivity, and improved neuroblast injury induced by d -galactose treatment. In addition, we also found that decreased immunoreactivities and protein levels of antioxidants including superoxide dismutase and catalase induced by d -galactose treatment were significantly recovered by 50 mg/kg TPM treatment in the mice hippocampal DG ( P < 0.05). In conclusion, our present results indicate that TPM can ameliorate neuroblast damage and promote cell proliferation and neuroblast differentiation in the hippocampal DG via increasing SODs and catalase levels in the d -galactose mice.
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JOURNAL OF PHARMACOLOGICAL SCIENCES
·
2014
·
IF 2.114
Jing Lu,Duanping Sun,Si Gao,Ying Gao,Jiantao Ye,Peiqing Liu
Autophagy is a highly regulated and multi-step biological process that serves to remove damaged cytoplasmic components and organelles. It has been suggested that the activation of autophagy may be a promising therapeutic strategy for cancer treatment by triggering cell death. In this study, we reported that cyclovirobuxine D (CVB-D), an alkaloid component in a traditional Chinese herb, could induce autophagy in the MCF-7 human breast cancer cell line. CVB-D inhibited the viability of MCF-7 cells in a concentration- and time-dependent manner. Activation of autophagy was characterized by transmission electron microscopy, monodansylcadaverine staining, and expression of autophagy marker microtubule-associated protein 1 light chain 3 (LC3). After CVB-D treatment, a clear accumulation of autophagosomes was observed accompanied with elevated LC3 fluorescent puncta. Western blot analysis revealed that CVB-D significantly promoted the conversion from LC3-I to LC3-II and the expression of autophagy-related protein 5 (ATG5), which are both essential for autophagosome formation. On the other hand, CVB-D–induced autophagy and decrease in cell viability could be blocked by 3-methyladenine, a well-established autophagy inhibitor. Moreover, CVB-D attenuated the phosphorylation of Akt and mTOR, two pivotal suppressors in autophagy pathways. These findings shed new light on the pharmacological actions and mechanism of CVB-D and may support the potential utility of autophagy inducers in cancer treatment.
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JOURNAL OF CHEMICAL NEUROANATOMY
·
2020
·
IF 2.353
Hongqing Liu,Shiqi Li,Cuizhu Yang,Hui Jia,Ziting Gu,Xing Tu,Sumin Tian,Jing Liu,Guoying Li,Yuxin Ma
The senile plaque formed by β-amyloid (Aβ) deposition in the brain is one of the main pathological features of Alzheimer’s disease (AD), and the c-Jun N-terminal kinase (JNK) signaling pathway plays an important role in the pathogenesis of AD. This study aimed to investigate that D-serine may ameliorate motor and cognitive impairment in Aβ injected mice by inhibiting JNK signaling pathway. Firstly, Kunming mice were injected intrahippocampally with Aβ 1-42 to build AD model. The mice were injected intraperitoneally with saline, D-serine, D-amino acid oxidase (DAAO), and Sodium benzoate (BE) for 10 consecutive days, respectively. Subsequently, the motor and cognitive functions of mice were detected by behavioral tests. The silver staining and immunohistochemical methods were used to detect the distributions of Aβ in the hippocampus of mice. 18 F-2-Fluro-D-deoxy-glucose positron emission tomography/computed tomography ( 18 F-FDG PET/CT) scans were performed to detected glucose metabolism of Aβ 1-42 induced lesions. The expressions of relative JNK factors were detected by immunohistochemistry and Western blot methods. These results showed that Aβ severely impaired the motor and memory abilities of mice. The expressions of glial fibrillary acidic protein (GFAP), tumor necrosis factor (TNF-α), N-methyl-D-aspartate receptor 1 (NMDAR1), phospho-JNK (p-JNK), p-c-Jun and activating transcription factor 2 (ATF2) increased significantly. After D-serine treatment, the abilities of movement and memory of mice were improved, and the clearance rate of Aβ was accelerated. The expressions of GFAP, TNF-α, NMDAR1, p-JNK, p-c-Jun and ATF2 decreased significantly. DAAO and BE were administered to further validate these results. Therefore, this study showed that D-serine could alleviate the cognitive impairment of Aβ 1-42 injected mice by inhibiting JNK signaling pathway. These results provide more evidences for the effect of D-serine on AD and relevant mechanism to treat AD.
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ENERGY & FUELS
·
2020
·
IF 3.421
Chunmei Deng,Mingyuan Zhu
In this research, we have prepared a new type of multistage aperture nitrogen-doped carbon materials [CN-x, (x = 600, 700, 800, 900, 1000)] by one-step carbonization at different calcination temperatures. The structure of the CN-x is characterized by X-ray diffraction, X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, Brunauer–Emmett–Teller, Scanning electron microscopy, and Raman spectroscopy. The effects of pore structure, nitrogen morphology, and defects on the adsorption properties are analyzed. We have also studied the desulfurization activity of CN-x for dibenzothiophene (DBT). Experimental data show that CN-900 has very good adsorption desulfurization activity, exhibiting a theoretical Langmuir adsorption capacity of up to 37.28 mg/g. When the DBT concentration is 100 ppm, CN-900 reaches a DBT removal rate of over 99%. The effects of calcination temperature and nitrogen content on the adsorption properties of the CN-x are examined. The experimental results show that the nitrogen content is positively correlated with the adsorption performance. The cheap and readily available d-(+)-glucosamine hydrochloride is used as the precursor of the CN-x, and the adsorbent prepared by a simple one-step carbonization method has good application prospects in the desulfurization process.
研究领域:
材料科学
·
环境工程
·
化学工程
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Medicinal Chemistry
·
2017
·Vol. 13
·期 8
·753-760
·
IF 2.331
Guo Ping,Xie Zixin,Zhang Huan,Zhang Zaikui,Han Chao,Cheng Donghua,Lin Dan,Zhang Yuan,Wang Xuebao,Guo Xin,Ye Faqing
Background: Thienopyrimidinone is a newly designed, selective fibroblast growth factor receptor 1 (FGFR1) inhibitor with an excellent anticancer effect. Objective: The goal of the present study was to design and synthesize better FGFR1 inhibitors through modifications of the lead
compound thienopyrimidinone. Methods: In the present study, a series of C-2 substituted derivatives of thienopyrimidinone, namely L1-L16, were synthesized, and their inhibitory effects on FGFR1 were evaluated. The anti-proliferative activities of these compounds were assessed by MTT assay.
Results: Among the novel derivatives, L11 was found to exert remarkable FGFR1 inhibitory activity (79.93% at 10 μM) and anti-proliferative activity, with IC50 values of 2.1, 2.5, and 3.5 µM in the FGFR1-overexpressing cell lines, H460, HT-1197, and B16F10, respectively.
Conclusion: Our newly synthesized thienopyrimidinone derivatives may be candidate FGFR1 inhibitors for future development as novel anticancer agents.
研究领域:
药理学
·
肿瘤学
·
药物化学
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