Materials Science & Engineering C-Materials for Biological Applications
·
2019
·
IF 4.959
Na Wen,Shaoyu Lü,Xiubin Xu,Piao Ning,Zengqiang Wang,Zinan Zhang,Chunmei Gao,Yongqi Liu,Mingzhu Liu
Various glucose-sensitive drug delivery platforms have been developed recently to treat diabetes. However, there is much less work has been reported on treatment of diabetes and vascular diabetes complications simultaneously. In this work, a novel polysaccharide-based micelle-hydrogel synergistic therapy system was fabricated to address this limitation. Zwitterionic dialdehyde starch-based micelles (SB-DAS-VPBA) were synthesized via single electron transfer-living radical polymerization (SET-LRP). Hydrophilic segment sulfobetaine (SB) and hydrophobic segment 4‑vinylphenylboronic acid (VPBA) were grafted to the dialdehyde starch (DAS) backbones. Then, chitosan/dialdehyde starch derivatives (CS/SB-DAS-VPBA) micelle-hydrogel was synthesized by Schiff-base bonds. Insulin and nattokinase were loaded to obtain the micelle-hydrogel synergistic therapy system. In vitro drug delivery and blood clots dissolution behaviors were determined. Results suggest that the micelle-hydrogel synergistic therapy system not only possesses glucose-responsive insulin delivery property, but also provides good thrombolytic capacity. Thus, this micelle-hydrogel synergistic therapy system can be used as a platform for diabetes and vascular diabetes complications treatment.
研究领域:
生物材料学
·
药物递送系统
·
糖尿病治疗
✓ 已验证
DOI
高影响因子
COLLOIDS AND SURFACES B-BIOINTERFACES
·
2017
·
IF 3.887
Shao-Fei Zhang,Chunmei Gao,Shaoyu Lü,Jiujun He,Mingzhu Liu,Can Wu,Yijing Liu,Xinyu Zhang,Zhen Liu
Nattokinase (NK) has been used as a new generation thrombolytic drug, due to its high safety, low cost and low side effects. However, it is sensitive to external environment and may lose the enzyme activity easily. Peptide dendrimer possesses functional groups on its surface, adjustable sizes, biodegradability, biocompatibility, and low toxicity, which could be used as ideal carrier for drug protection and delivery. Demonstrated for the first time in this paper, a PEGylated dendrimer (G n -PEG-G n ) composed of polyglutamic acid is designed and synthesized as delivery platform of NK for thrombus treatment. A panel of PEGylated dendrimers with three different generations of 2, 3, 4 was prepared to investigate the effect of dendrimer architecture on the properties and therapeutic efficacy of the resultant NK-loaded delivery systems in terms of the morphology, dimension and enzyme activity. The results demonstrated that the NK-loaded G 3 -PEG-G 3 (G 3 -PEG-G 3 /NK ratio of 6/1), of all the formulations, displayed the optimal enzyme activity for dissolving thrombus in vitro , thus offering great potential for the treatment of thrombus.
研究领域:
生物材料
·
药物递送
·
医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOPROCESS AND BIOSYSTEMS ENGINEERING
·
2025
·
IF 3.6
Zheng Jiawen,Sun Yaping,Liao Yunyu,Qin Peng,Che Rongzhen,Zhao Jing-yi,Xiao Zijun
Nattokinase (NK) is one of the most important functional components in natto, but its content is low. In this study, the fermentation conditions using Bacillus subtilis JZ08-02 for high-yield NK production were investigated, and the residual bacterial pellets were used to prepare a natto starter. Batch fermentation of NK was conducted using a 5 L fermenter, and soybean milk and glucose were used as the substrates. When the stirring speed was increased from 450 to 650 rpm with air supply at 1.0 vvm, NK was increased from 4859 ± 142 to 12,294 ± 226 IU/mL. When pure oxygen was supplied, 15,013 ± 550 IU/mL of NK was obtained. When fed-batch fermentation was conducted, the titer was further elevated to 18,014 ± 112 IU/mL, which was increased by about 76% compared with the previous result. The experimental findings revealed that aeration control and nutrient feeding regimens exerted pronounced effects on NK productivity during submerged fermentation. The crude enzyme supernatant was obtained by centrifugation and the precipitate was collected. With optimized protectant, the bacterial pellets were freeze-dried with 90.1% cell survival rate. Using economical and edible feedstocks, this study achieved a significant enhancement in NK fermentation yield via oxygen-enriched fed-batch cultivation. At the same time, a natto starter was prepared as a by-product using the residual cell waste.
研究领域:
微生物学
·
生物工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
POLYMERS FOR ADVANCED TECHNOLOGIES
·
2019
·Vol. 30
·期 9
·2353-2360
·
IF 2.162
Mengjie Huang,Shaoyu Lü,Yanzheng Ji,Zengqiang Wang,Shao-Fei Zhang,Taomei Qi,Jia Yan,Tao Li,Yongming Liu,Mingzhu Liu
Thrombosis has become one of the major diseases harmful to human health. The new-generation thrombolytic drug nattokinase (NK) has been used in the treatment of thrombosis because of its high safety, small side effects, and good thrombolytic effect. However, its application is limited because of its short half-life and inactivation in external environments. Therefore, it is necessary to retain the enzyme activity of NK to improve its environmental stability. In this study, NK was decorated by bonding peptide dendrimers to improve its enzymatic activity. Polyglutamic acid dendrimers of generation 2 were firstly synthesized, and then the targeting molecule arginine-glycine-aspartic acid (RGD) was linked to dendrimers by reaction of carboxyl and amino groups. Subsequently, polyglutamic dendrimer particles were bonded on the periphery of NK by chemical reaction and were used as carriers for NK. The carriers were characterized by Fourier transform infrared (FTIR) and thermogravimetric (TG) analysis, and their size and morphology were evaluated. Their enzyme activity at different pH and temperature was determined, and improved NK activity was achieved after bonding with dendrimers under acid and alkaline conditions and high temperature. Their thrombolytic properties were evaluated by in vitro and in vivo thrombolytic experiments. Effective thrombolytic effect was observed. This study indicated that the NK carrier prepared by chemical bonding may have applications in the treatment of thrombosis.
研究领域:
生物化学
·
药学
·
医学
✓ 已验证
产品关联
DOI
高影响因子
Food Bioscience
·
2025
·
IF 5.9
Yongjie Zhou,Zixin Fu,Peipei Dou,Chen Yu,Xianyou Dong,Hui Hong,Yongkang Luo,Yuqing Tan
Nattokinase (NK), an alkaline serine protease, is recognized for its cardiovascular benefits through the modulation of blood coagulation and fibrinolytic pathways. Atherosclerosis (AS), a primary cause of cardiovascular disease (CVD) worldwide, is responsive to dietary interventions. However, studies investigating NK’s digestion, absorption, and mechanisms in mitigating AS remain limited. This study evaluated the effects of high-dose NK treatment (H-NK, 900 FU/kg body weight) administered over six weeks in ApoE -/- mice, which resulted in significant reductions in atherosclerotic lesions. In vitro digestion simulations showed that 25.8% of NK’s enzymatic activity was retained post-digestion, and thrombolysis assays demonstrated a thrombolytic efficacy of 30%-40% at concentrations ≥ 5,000 μg/mL, regardless of clot age. Absorption experiments using a rat everted intestinal sac model revealed a small intestine absorption rate of 37.0%, with the absorbed peptides being predominantly hydrophobic, uncharged, and shorter than 10 amino acids. Proteomic analysis of mouse liver tissues identified that high-dose NK down-regulated PXDN (peroxidasin) and PNLIP (pancreatic lipase), proteins involved in promoting lipid oxidation, and intestinal lipid absorption. These proteomic changes enhanced lipid metabolism, reduced lipid peroxidation, and alleviated AS through the "Lipid and Atherosclerosis" pathway. Overall, this study provides a deeper understanding of NK’s biological activity and supports its potential as a novel dietary supplement for the management of CVD.
研究领域:
心血管疾病研究
·
营养学
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART A
·
2017
·Vol. 106
·期 2
·440-449
·
IF 3.076
Can Wu,Chunmei Gao,Shaoyu Lü,Xiubin Xu,Na Wen,Shaofei Zhang,Mingzhu Liu
Thrombotic disease has become one of the leading causes of mortality among humans globally. Nattokinase (NK), a novel thrombolytic agent, has attracted the attention of researchers. However, NK is a serine protease that is vulnerable to environmental effects resulting in its inactivation. In this study, polylysine dendrimer (PLLD) was synthesized through divergence-convergence method, and a series of NK/PLLD nanocomposites with different molar ratio was prepared. In addition, NK was successfully incorporated into the cores of PLLD G4 through hydrogen bonds and van der Waals forces. In NK/PLLD nanocomposites, when the molar ratio of NK to PLLD is 1:30, a high relative enzyme activity level (up to 117%) was achieved and was more stable at different temperatures and pH than free NK. In in vitro thrombolysis experiment, compared with free NK, NK/PLLD nanocomposites could control the release of NK. The thrombolysis rate of NK/PLLD nanocomposites reached 50% at 12 h, which can effectively avoid other complications such as hemorrhage. Interestingly, NK/PLLD nanocomposites with positive charge can penetrate into the negatively charged thrombus through electrostatic interaction, thus providing a good thrombolytic effect. Hemolysis and MTT experiments show that PLLD nanomaterials can serve as ideal carriers of protein drugs. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 440–449, 2018.
研究领域:
生物材料学
·
医学
·
药剂学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
PROCESS BIOCHEMISTRY
·
2024
·
IF 3.7
Xilin Huang,Xuli Gao,Junbao Huang,Yani Luo,Wei Tao,Mingyu Guo,Yongyuan Liu,Jing Wu,Yu Chen,Yan Liu
A recombinant strain (BS168-Δ bdhA ) that increased co-production of MK-7 and NK was constructed by knocking out the 2,3-butanediol dehydrogenase gene ( bdhA ) of Bacillus subtilis 168 to reduce the carbon spillover. The results showed that the 2,3- butanediol content in the recombinant strain was reduced by 64% to 2.76 g/L. The recombinant strain had an MK-7 production of 30.6 mg/L and an NK activity of 6.5 FU/mL. RNA-Seq analysis showed that: bdhA knockdown blocked the carbon flux of 2,3-butanediol and promoted glycerol uptake, and the carbon flux flowed more to the MK-7 and NK synthesis pathways. The negative regulator of NK, codY , was down-regulated by 2.19-fold. secA , which is responsible for the energy supply of transmembrane proteins, was down-regulated by 0.37-fold, the Tat secretion pathway was up-regulated by 2.8-fold and 0.5-fold for tatAD and tatC , respectively. This study revealed the mechanism of co-production of MK-7 and NK in recombinant strains for the first time and revealed potential molecular targets to promote the co-production of MK-7 and NK, which will provide a new strategy for metabolic engineering to increase the production of MK-7 and NK.
研究领域:
微生物学
·
分子生物学
·
代谢工程
✓ 已验证
产品关联
DOI
高影响因子
RSC Advances
·
2014
·Vol. 4
·期 52
·27422-27429
·
IF 3.708
Chen Chen,Haogang Duan,Chunmei Gao,Mingzhu Liu,Xin'an Wu,Yuhui Wei,Xinyu Zhang,Zhen Liu
Cardiovascular disease (CVD) has become the leading cause of death globally. Nattokinase (NK), as a novel thrombolytic agent, has gained more and more attention. However, NK is susceptible to chemical oxidation, and subsequent inactivation denaturation. Modification of an enzyme to improve its stability usually brings about falling enzymatic activity. In this study, folic acid modified chitosan (CS-FA) was synthesized, and a series of NK/CS-FA complexes with different mass ratios were prepared. In vitro thrombolysis experiments indicated that NK/CS-FA showed obvious advantages in its fibrinolysis activity and stability. With the NK : CS-FA mass ratio of 100 : 1 (molar ratio of NK : chitosan pyranose ring = 3 : 5), NK/CS-FA 100 : 1 exhibited the best fibrinolysis activity and the thrombi dissolved completely within 17 hours at a constant rate, while the un-modified NK dissolved only 35 wt% of the thrombus in the same duration. NK/CS-FA 100 : 1 retained more than 53% enzymatic activity at 80 °C or pH 2, as a comparison, only 17% enzymatic activity was left for un-modified NK under the same conditions.
研究领域:
生物化学
·
医学
✓ 已验证
产品关联
DOI
高影响因子
Materials Science & Engineering C-Materials for Biological Applications
·
2020
·
IF 5.88
Mengjie Huang,Yanzheng Ji,Jia Yan,Taomei Qi,Shao-Fei Zhang,Tao Li,Shaoyu Lü,Yongming Liu,Mingzhu Liu
Thrombotic complications turn into the second leading cause of death in colon cancer patients due to the hypercoagulable state caused by malignancy. Therefore, it is necessary to treat colon cancer and its thrombosis complications simultaneously. Herein, a nano polymer conjugate based on disulfide cross-linked low-generation peptide dendrimers was developed to treat colon cancer and its thrombotic complications. First, two-generation polyglutamic acid dendrimer was bonded to nattokinase (NK) and then cross-linkers containing disulfide linkages were used to obtain polymer conjugates (NK-G 2 ) n . Then doxorubicin (Dox) was encapsulated. The system can release drugs sequentially due to the dissociation of the polymer conjugates. In vitro thrombolytic experiments exhibited a significant thrombolysis ability of (NK-G 2 ) n . The toxicity and cellular uptake tests on HCT116 cells showed that Dox loaded polymer conjugates had good endocytosis ability and anti-cancer effect. Therefore, this drug delivery system will be a promising strategy to the combined treatment of colon cancer and thrombotic complications.
研究领域:
肿瘤学
·
药剂学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART A
·
2018
·Vol. 106
·期 6
·1687-1696
·
IF 3.231
Shao-Fei Zhang,Shaoyu Lü,Chunmei Gao,Jiandong Yang,Xiang Yan,Tao Li,Na Wen,Mengjie Huang,Mingzhu Liu
Thrombotic events affect many individuals in a number of ways, all of which can cause significant morbidity and mortality. Nattokinase (NK), as a novel thrombolytic drug, has been used for thrombolytic therapy. It not only possesses plasminogen activator activity, but also directly digests fibrin through limited proteolysis. However, it may undergo inactivation and denaturation in the harsh external environment. In this study, a multiarm-polyethylene glycol-polyglutamic acid peptide dendrimer was fabricated and used as a carrier for NK protection and delivery. Different arm numbers of polyethylene glycol-polyglutamic acid peptide dendrimers ( x -PEG(G 3 ) x , x = 2, 4, 6, 8) were designed, prepared, and characterized by 1 H NMR and FTIR. Then, x -PEG(G 3 ) x were loaded with NK to form nanocomposites. Their size and morphology were determined by dynamic light scattering and transmission electron microscopy. Enzyme activity was evaluated via UV–Vis absorbance spectra, fluorescence spectra, circular dichroism spectra, and zeta potential measurements. The study reveals that the obtained x -PEG(G 3 ) x /NK nanocomposites possess high enzyme activity. In addition, the nanocomposites show increased viability of rat macrophage cells, and excellent thrombolysis ability in vitro and in vivo . This work establishes a multiarm-polyethylene glycol-polyglutamic acid peptide dendrimer with potential application in NK carrier and thrombolytic therapy. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1687–1696, 2018.
研究领域:
生物医学材料
·
药物递送系统
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF FOOD BIOCHEMISTRY
·
2019
·Vol. 43
·期 5
·e12820
·
IF 1.358
Na Guo,Yi-Wei Jiang,Xin-Rui Song,Yan-Yan Li,Zhi-Ming Liu,Yu-Jie Fu
In the present investigation, fibrinolytic Ginkgo seeds were produced by solid-state fermentation (SSF) with Bacillus natto strains, and some parameters of the fermentation processes were investigated. Under optimal fermentation conditions, the fibrinolytic activity of Ginkgo seeds reached 3,682 ± 43 IU/g with the fermentation parameters of relative humidity 80%, initial water content 73%, fermentation temperature 38°C, inoculation volume 18%, and fermentation time 38 hr, respectively. The fermented Ginkgo seeds possessed a superior potential for the production of Nattokinase. What's more, the fermented Ginkgo seeds possessed higher total flavonoid and lower ginkgolic acids contents, which could enhance bioactivity and guarantee food safety. Sensory evaluations indicated that Ginkgo seeds produced by SSF could also be consumed as a kind of popular food. Practical applications Fermented food is popular in countries. It can not only improve the sensory properties of the products, reduce undesirable constituents, and make nutrients easily absorbable, but also improve the nutritional properties. Ginkgo biloba L is one of the oldest species that has existed on earth for more than 200 million years. However, the application of Ginkgo seeds has been limited because of the ginkgolic acids. In a previous study, immobilized Bacillus natto acted upon Ginkgo seeds to enhance the bioactivity and safety of fermented Ginkgo seeds. However, separating the fermented Ginkgo seeds from the liquid needs a large amount of energy. The solid-state fermentation of Ginkgo seeds is a good choice to produce functional Ginkgo seed products.
研究领域:
食品科学
·
微生物学
·
营养学
·
植物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD RESEARCH INTERNATIONAL
·
2022
·
IF 7.425
Yejun Zhong,Lei Yang,Ziyi Zhu,Hongyu Chen,Chengmei Liu,Taotao Dai,Er Sheng Gong
The protective effect of ovalbumin-flavonoids (naringenin, genistein, naringin, puerarin, and daidzein) hydrogels on the thrombolytic activity and stability of nattokinase were investigated. The results suggested that flavonoids promoted the gelation of ovalbumin solution, which was not able to form hydrogel at the same concentration upon heating. The nattokinase/ovalbumin-naringenin hydrogel had the strongest hardness and springiness. All nattokinase/ovalbumin-flavonoids hydrogels were more elastic than viscous. After in vitro digestion, the thrombolytic capacities of all nattokinase/ovalbumin-flavonoids hydrogels were significantly higher than that of free nattokinase. Moreover, nattokinase/ovalbumin-flavonoids hydrogels showed higher thermal and pH stability than free nattokinase. Fluorescence spectroscopy and molecular docking analysis revealed that the main interactions between nattokinase and daidzein, nattokinase and genistein were mainly hydrogen bond, while the main interactions between nattokinase and naringin, nattokinase and puerarin, nattokinase and naringenin were hydrophobic interaction. This research suggested that nattokinase/ovalbumin-flavonoids had great potential for applications in the treatment of thrombus.
研究领域:
药学
·
食品科学
·
生物技术
✓ 已验证
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）