WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
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2024
·Vol. 41
·期 1
·1-12
·
IF 4
Li Yang-Nan,Zhu Chen-Yuan,Xu Chen-Lu,Yu Shen,Huan Tong,Zhang Ye-Wang
A heparinase III ( Ns Hep-III) from Niabella sp . was identified, cloned, and expressed as soluble form in E. coli BL21 (DE3). With heparin as substrate, the maximum activity of Ns Hep-III of 90.0 U·mg − 1 was achieved at pH 7.1 Tris-HCl (containing 15 mM Mg 2+ ) and 30 o C. The half-life of Ns Hep-III was determined to be 5 h at 30 o C. The interactions between Ns Hep-III and substrates were studied by molecular docking. The combination of Ns Hep-III and a heparinase I from Bacteroides eggthii ( Be Hep-I) was employed to cleave heparin. Analysis of the enzymatic products of Ns Hep-III by SAX-HPLC showed seven different modified disaccharides, indicating that Ns Hep-III has a wide range of substrate specificity. The results of GPC analysis demonstrated that the average molecular weight of the product of heparin cleavage by the combination of Ns Hep-III/ Be Hep-I was reduced to 3969 Da, which accounted for 90% of all the components, and complied with the requirements of the European Pharmacopoeia. Ns Hep-III has notable activity and efficiency in cleaving heparin, which is potentially useful for the industrial production of low molecular weight heparin.
研究领域:
生物化学
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药学
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高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2024
·
IF 5.7
Chen-Lu Xu,Chen-Yuan Zhu,Yang-Nan Li,Jian Gao,Ye-Wang Zhang
A novel heparinase III from Pedobacter schmidteae (PsHep-III) with high activity and good stability was successfully cloned, expressed, and characterized. PsHep-III displayed the highest specific activity ever reported of 192.8 U mg–1 using heparin as the substrate. It was stable at 25 °C with a half-life of 323 h in an aqueous solution. PsHep-III was employed for the depolymerization of heparin, and the enzymatic hydrolyzed products were analyzed with gel permeation chromatography and high-performance liquid chromatography. PsHep-III can break glycosidic bonds in heparin like →4]GlcNAc/GlcNAc6S/GlcNS/GlcNS6S/GlcN/GlcN6S(1 → 4)ΔUA/ΔUA2S[1 → and efficiently digest heparin into seven disaccharides including N-acetylated, N-sulfated, and N-unsubstituted modification, with molecular masses of 503, 605, 563, 563, 665, 360, and 563 Da, respectively. These results indicated that PsHep-III with broad substrate specificity could be combined with heparinase I to overcome the low selectivity at the N-acetylated modification binding sites of heparinase I. This work will contribute to the application of PsHep-III for characterizing heparin and producing low-molecular-weight heparin effectively.
研究领域:
生物化学
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分子生物学
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制药工程
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BIOSENSORS & BIOELECTRONICS
·
2016
·
IF 7.476
Yihui Hu,Wenjing Guo,Yubin Ding,Hanjun Cheng,Hui Wei
The detection of heparin (Hep) and its contaminant oversulfated chondroitin sulfate (OSCS) is of great importance in clinics but remains challenging. Here, we report a sensitive and selective time-resolved luminescence (TRL) biosensing system for Hep by modulating the photoluminescence of Tb 3+ with guanine-rich ssDNA and Hep-specific AG73 peptide (RKRLQVQLSIRT). With the developed system, Hep including both unfractionated Hep (UFH) and the low molecular weight Hep (LMWH) has been successfully detected with a satisfactory detection limit. Owing to the highly specific interaction between Hep and AG73 peptide, major interfering substances in Hep detection, such as Hep analogs of chondrotin sulfate (Chs) and hyaluronic acid (HA), did not interfere with Hep detection. The established TRL sensing system was then successfully used for monitoring Hep metabolism in living rats by microdialysis . Moreover, the proposed TRL sensing system was further applied to analyze OSCS contaminant in Hep with heparinases treatment by exploring the inhibition effects of OSCS on the activity of heparinases. As low as 0.002% of OSCS in Hep was identified.
研究领域:
分析化学
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生物医学工程
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药学
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SENSORS AND ACTUATORS B-CHEMICAL
·
2016
·
IF 4.758
Dan Yan,Yu He,Yili Ge,Gongwu Song
A novel fluorescence “turn on-off” method for sensitive and selective detection of heparin and heparinase I was developed based on near infrared Polyethyleneimine Capped Ag 2 S Quantum Dots (PEI-Ag 2 S QDs). The fluorescence “turn-on” arose from aggregation-induced emission enhancement (AIEE) characteristics of PEI-Ag 2 SQDs induced by electrostatic interaction between PEI-Ag 2 SQDs and heparin. Heparinase I, which can specifically hydrolyze heparin into short fragments would de-aggregate PEI-Ag 2 S QDs/heparin system and lead to fluorescence “turn-off”. Under optimal conditions described, the fluorescence enhancement at 685 nm was linearly related to the concentration of heparin and heparinase I in the range of 0.069–2.275 μM and 0–103 ng mL −1 with the relative correlation coefficient of R 2 = 0.997 and 0.99 and detection limit as low as 6 nM and 0.5 ng mL −1 , respectively. The activity of the heparinase I was further investigated using steady-state kinetic for the reaction. In ddition, the proposed method was successfully utilized in serum samples, which presaged more opportunities for application in related biochemical and biomedical researches.
研究领域:
生物化学
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医学检测
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纳米技术
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2019
·
IF 4.784
Rui Jiang,Song Zhao,Langkun Chen,Maoyu Zhao,Wenjing Qi,Wensheng Fu,Lianzhe Hu,Yan Zhang
In this research, a new DSA (Distyryl-anthracene) derivative with four carboxyl groups was designed and synthesized. This molecule exhibits aggregation-induced emission property (AIE). With the AIE character, a convenient and sensitive fluorescent probe for the detection of protamine , heparin and heparinase has been developed. The protamine can directly induce the aggregation of probe, which caused by electrostatic attraction. In this way, the turn-on detection of protamine is achieved, and the detection limit is as low as 30 ng mL −1 . When heparin appears, the probes will be redisperse in solution, which causes a decrease in fluorescence intensity. Besides, this method also shows good selectivity and sensitivity, and the linear range of heparin is from 0.08 to 8 μg mL −1 with detection limit of 37 ng mL −1 . After hydrolyzing heparin by heparinase, the probes rebind with protamine and the fluorescence enhance. The fluorescence enhancement was linearly related to the concentration of heparinase in the range of 0.02–2.0 μg mL −1 and detection limit as low as 143.7 ng mL −1 . In addition, the results exhibited that the recovery percentage of heparinase in bovine samples reached to 96–101%.
研究领域:
生物化学
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分析化学
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高影响因子
BIOMACROMOLECULES
·
2015
·
IF 5.75
Chuanxiong Nie,Lang Ma,Chong Cheng,Jie Deng,Changsheng Zhao
Combining the advantages of the fibrous nanostructure of carbon nanotubes (CNTs) and the bioactivities of heparin/heparin-mimicking polyanions, functional nanofibrous heparin or heparin-mimicking multilayers were constructed on PVDF membrane with highly promoted endothelialization and antithrombogenic activities. Oxidized CNT (oCNT) was first functionalized with water-soluble chitosan (polycation), then enwrapped with heparin or a typical sulfonated heparin-mimicking polymers (poly(sodium 4-styrenesulfonate-co-sodium methacrylate)) to construct the multilayers. Then, the surface-deposited multilayers were constructed via electrostatic layer-by-layer assembly of the functionalized oCNTs. The scanning electron microscope and atom force microscope images confirmed that the coated multilayers exhibited nanofibrous and porous structure. The live/dead cell staining and cell viability assay results indicated that the coated nanofibrous multilayers had excellent compatibility with endothelial cells. The cell morphology observation further confirmed the promotion ability of surface endothelialization due to the coated heparin/heparin-mimicking multilayers. Further systematical evaluation on blood compatibility revealed that the surface heparin/heparin-mimicking multilayer-coated membranes also had significantly improved blood compatibility including restrained platelet adhesion and activation, prolonged blood clotting times, and inhibited activation of coagulation and complement factors. In summary, the proposed nanofibrous multilayers integrated endothelialization and antithrombogenic properties; meanwhile, the heparin-mimicking coating validated comparable performances as heparin coating. Herein, it is expected that the surface coating of nanofibrous multilayers, especially the facilely constructed heparin-mimicking coating, may have great application potential in biomedical fields.
研究领域:
生物医学工程
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材料科学
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细胞生物学
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高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
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2021
·Vol. 22
·期 8
·4077
·
IF 5.924
Xiao-Lei Qiu,Zi-Rui Fan,Yang-Yang Liu,Ding-Fu Wang,Shi-Xin Wang,Chun-Xia Li
A self-nanoemulsifying drug delivery system (SNEDDS) was developed to enhance the absorption of heparin after oral administration, in which heparin was compounded with phospholipids to achieve better fat solubility in the form of heparin-phospholipid (HEP-Pc) complex. HEP-Pc complex was prepared using the solvent evaporation method, which increased the solubility of heparin inn-octanol. The successful preparation of HEP-Pc complex was confirmed by differential scanning calorimetry (DSC), Fourier-transform infrared (FT-IR) spectroscopy, NMR, and SEM. A heparin lipid microemulsion (HEP-LM) was prepared by high-pressure homogenization and characterized. HEP-LM can enhance the absorption of heparin after oral administration, significantly prolong activated partial thromboplastin time (APTT) and thrombin time (TT) in mice, and reduce fibrinogen (FIB) content. All these outcomes indicate that HEP-LM has great potential as an oral heparin formulation.Keywords:heparin;heparin–phospholipid complex;self-nanoemulsifying drug delivery system;oral absorption
研究领域:
药剂学
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生物医学工程
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PubMed
高影响因子
Coatings
·
2023
·Vol. 13
·期 7
·1248
·
IF 3.4
Jincan Lei,Haiyan Wang,Qi Dang,Xiang Wang,Chungong Li,Jing Huang,Jingzhou Hou,Kejing Fang,Shixian Zhao
Activated carbon (AC) is an effective adsorbent for creatinine removal in hemoperfusion. However, the hemocompatibility and adsorption capacity of AC was required to be improved further. Heparin has different anticoagulant mechanisms due to its different molecular weights. Thus, it was necessary to study the surface modification with unfractionated heparin (UFH) or low-molecular-weight heparin (LMWH) on improvement of hemocompatibility and adsorption. In this study, UFH and LMWH were, respectively, grafted on AC through polyethyleneimine as an intermediate layer. The modification of AC regarding morphology, mechanical strength, and pore structure was characterized by X-ray photoelectron spectroscopy (XPS), scanning electron microscope (SEM), texture analyzer (TA), and surface area analyzer. It was found that, compared with AC, the morphology and mechanical strength of AC-UFH and AC-LMWH could be well maintained, but the specific surface area was decreased due to the grafting of macromolecules. Furthermore, AC-UFH and AC-LMWH showed better hemocompatibility on protein adsorption, clotting time, and platelet activation compared with AC, in which AC-LMWH had lower fibrinogen adsorption and longer clotting time than AC-UFH. In addition, it was found that AC, AC-UFH, and AC-LMWH had no significant effect on blood cell composition. Finally, the adsorption capacity of adsorbents for creatinine was evaluated. Although there was no significant difference between AC-UFH and AC-LMWH, it was found that heparin could be interacted with creatinine to enhance the adsorption capacity when compared with polyethyleneimine-modified AC. This study deepened the understanding of anticoagulation of heparinized surface and provided a theoretical basis for adsorption in hemoperfusion.Keywords:heparin;hemocompatibility;activated carbon;adsorption
研究领域:
生物医学工程
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材料科学
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血液透析
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CHROMATOGRAPHIA
·
2015
·Vol. 78
·期 11
·833-837
·
IF 1.411
Zhang Mingyu,Qiu Danye,Kang Jingwu
A simple, sensitive and environmentally friendly capillary electrophoresis method for the determination of free sulfate anions in heparin and low-molecular-weight heparin with indirect UV detection is reported. The background electrolyte consists of 20 mM nitrate (pH 5.5) containing 0.2 mM CTAB. The use of nitrate as the probe anion is based on the consideration that nitrate has a very close electrophoretic mobility to that of sulfate, leading to a very narrow peak shape. Moreover, the apparent molar absorptivity of nitrate at 202 nm (8413 L mol −1 cm −1 ) is much higher than that of the commonly used probe anion chromate at 254 nm (2400 L mol −1 cm −1 ). The combination of the narrow peak shape with the high apparent molar absorptivity of the probe ion improved the limit of detection by 2.4 times and the limit of quantitation by 3.2 times. The effect of various CE parameters on the separation of sulfate from chloride was investigated and optimized. The method displays linearity in the range from 0.05 to 20 mM. The RSD % for intraday ( n = 6), and interday ( n = 3) repeatability in terms of migration times and peak areas were all less than 3 %. The limit of detection ( S / N = 3) and limit of quantitation ( S / N = 10) for sulfate were determined as 18 and 53 µM, respectively. The method was successfully applied for the quantification of sulfate anions in heparin and low-molecular-weight heparins.
研究领域:
分析化学
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药学
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高影响因子
JOURNAL OF BIOMATERIALS APPLICATIONS
·
2021
·
IF 2.646
Qing Zhu,Ping Ye,Fang Guo,Yimen Zhu,Wenbin Nan,Zhaohua Chang
In this study, the surface of the covered stent was treated by plasma technology to introduce amino functional groups, and glutaraldehyde and heparin were successfully grafted to prepare a heparin-functionalized covered stent (HPLCS). The preparation parameters such as plasma treatment power, plasma treatment time, concentration of glutaraldehyde and heparin, and pH of heparin solution were studied in detail. The functionalized heparin covered stent can make the titer of heparin reach 1.23 ± 0.03 IU/cm2. In animal experiments, after implantation in pigs for 6 months, the titer of heparin can still reach 0.93 ± 0.05 IU/cm2. This work provides a good method for preparing heparin covered stent.
研究领域:
生物医学工程
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材料科学
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Polymers
·
2022
·Vol. 14
·期 12
·2470
·
IF 4.967
Jing Yang,Xujiao Tang,Qingqing Wu,Panpan Ren,Yishu Yan,Wei Liu,Chun Pan
Heparin has shown benefits in severe acute pancreatitis (SAP) therapy, but the underlying mechanisms were unknown. Extracellular high-mobility group protein-1 (HMGB-1) has been regarded as a central mediator contributing to inflammation exacerbation and disease aggravation. We hypothesized heparin attenuated the disease by targeting HMGB-1-related pathways. In the present study, the possible therapeutic roles of heparin and its non-anticoagulant derivatives, 6-O-desulfulted heparin and N-acylated-heparin, were determined on mouse models induced by “Two-Hit” of L-arginine. The compounds exhibited potent efficiency by substantially decreasing the pancreatic necrosis, macrophage infiltration, and serum inflammatory cytokine (IL-6 and TNF-α) concentration. Moreover, they greatly reduced the rapidly increasing extracellular HMGB-1 levels in the L-arginine injured pancreases. As a result, multiple organ failure and mortality of the mice were inhibited. Furthermore, the drugs were incubated with the RAW264.7 cells activated with damaged pancreatic tissue of SAP mice in vitro. They were found to inhibit HMGB-1 transfer from the nucleus to the plasma, a critical step during HMGB-1 active secretion from macrophages. The results were carefully re-examined with a caerulein and LPS induced mouse model, and similar results were found. The paper demonstrated heparin alleviated SAP independent of the anti-coagulant functions. Therefore, non-anticoagulant heparin derivatives might become promising approaches to treat patients suffering from SAP.Keywords:heparin;severe acute pancreatitis;HMGB-1;macrophage
研究领域:
药理学
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免疫学
·
炎症
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胃肠病学
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PubMed
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BIOSENSORS & BIOELECTRONICS
·
2015
·
IF 6.409
Meiding Yang,Jian Chen,Huipeng Zhou,Wenying Li,Yan Wang,Juanmin Li,Cuiyun Zhang,Chuibei Zhou,Cong Yu
A benzoperylene probe excimer emission in an aqueous buffer solution is observed for the first time, and a novel ratiometric fluorescence method based on the probe excimer emission for the sensitive detection of heparin and heparinase is demonstrated. A negatively charged benzoperylene derivative, 6-(benzo[ghi]perylene-1,2-dicarboxylic imide-yl)hexanoic acid (BPDI), was employed. A polycation , poly(diallyldimethylammonium) chloride (poly-DDA), could induce aggregation of BPDI through noncovalent interactions. A decrease of BPDI monomer emission and a simultaneous increase of BPDI excimer emission were observed. Upon the addition of heparin, the strong binding between heparin and poly-DDA caused release of BPDI monomer molecules, and an excimer–monomer emission signal transition was detected. However, after the enzymatic hydrolysis of heparin by heparinase, heparin was hydrolyzed into small fragments, which weakened the competitive binding of heparin to poly-DDA. Poly-DDA induced aggregation of BPDI, and a monomer–excimer emission signal transition was detected. Our assay is simple, rapid, inexpensive, sensitive and selective, which could facilitate the heparin and heparinase related biochemical and biomedical research.
研究领域:
分析化学
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生物化学
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生物医学工程
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PubMed
高影响因子
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