PROTEIN EXPRESSION AND PURIFICATION
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2022
·
IF 2.025
Chengyao Xia,Ding Li,Mengyi Qi,Yanxin Wang,Yue Zhang,Yiheng Yang,Zejia Hu,Xin Du,Yuqiang Zhao,Kuai Yu,Yan Huang,Zhoukun Li,Xianfeng Ye,Zhongli Cui
Chitosanases hydrolyze chitosan into chitooligosaccharides (COSs) with various biological activities, which are widely employed in many areas including plant disease management. In this study, the novel chitosanase Aq Csn1 belonging to the glycoside hydrolase family 46 (GH46) was cloned from Aquabacterium sp. A7-Y and heterologously expressed in Escherichia coli BL21 (DE3). Aq Csn1 displayed the highest hydrolytic activity towards chitosan with 95% degree of deacetylation at 40 °C and pH 5.0, with a specific activity of 13.18 U/mg. Product analysis showed that Aq Csn1 hydrolyzed chitosan into (GlcN) 2 and (GlcN) 3 as the main products, demonstrating an endo-type cleavage pattern. Evaluation of antagonistic activity showed that the hydrolysis products of Aq Csn1 suppress the mycelial growth of Magnaporthe oryzae and Phytophthora sojae in a concentration-dependent manner, and the inhibition rate of P. sojae reached 39.82% at a concentration of 8 g/L. Our study demonstrates that Aq Csn1 and hydrolysis products with a low degree of polymerization might have potential applications in the biological control of agricultural diseases.
研究领域:
生物技术
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微生物学
·
植物病理学
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DOI
PubMed
高影响因子
Marine Drugs
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2021
·Vol. 19
·期 11
·596
·
IF 5.118
Lin-Lin Zhang,Xiao-Hua Jiang,Xin-Feng Xiao,Wen-Xiu Zhang,Yi-Qian Shi,Zhi-Peng Wang,Hai-Xiang Zhou
(1) Background: Chitooligosaccharides (COS) have numerous applications due to their excellent properties. Chitosan hydrolysis using chitosanases has been proposed as an advisable method for COS preparation. Although many chitosanases from various sources have been identified, the cold-adapted ones with high stability are still rather rare but required. (2) Methods: A novel chitosanase named CsnY from marine bacteriumRenibacteriumsp. Y82 was expressed inEscherichia coli, following sequence analysis. Then, the characterizations of recombinant CsnY purified through Ni–NTA affinity chromatography were conducted, including effects of pH and temperature, effects of metal ions and chemicals, and final product analysis. (3) Results: The GH46 family chitosanase CsnY possessed promising thermostability at broad temperature range (0–50 °C), and with optimal activity at 40 °C and pH 6.0, especially showing relatively high activity (over 80% of its maximum activity) at low temperatures (20–30 °C), which demonstrated the cold-adapted property. Common metal ions or chemicals had no obvious effect on CsnY except Mn2+and Co2+. Finally, CsnY was determined to be an endo-type chitosanase generating chitodisaccharides and -trisaccharides as main products, whose total concentration reached 56.74 mM within 2 h against 2% (w/v) initial chitosan substrate. (4) Conclusions: The results suggest the cold-adapted CsnY with favorable stability has desirable potential for the industrial production of COS.Keywords:chitosanase;cold adaptation;chitooligosaccharide;Renibacteriumsp.
研究领域:
生物化学
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生物技术
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海洋生物学
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产品关联
DOI
PubMed
高影响因子
CARBOHYDRATE POLYMERS
·
2024
·
IF 10.7
Jinxin Lan,Yao Wu,Jiazhen Chen,Peng Wang,Hui Chen,Jinfeng Huang,Dongdong Lu,Changmei Lin,Xiaojuan Ma,Shilin Cao
As the most abundant renewable resource, cellulose fibers are potential candidates for use in health-protective clothing. Herein, we demonstrate a novel strategy for preparing cellulose fiber with prominent antibacterial and antiviral performance by the synergistic effect of amino groups and sulfonic acid groups. Specifically, guanylated chitosan oligosaccharide (GCOS) and N-sulfopropyl chitosan oligosaccharide (SCOS) were synthesized and chemically grafted onto cellulose fibers (CFs) to endow the fibers with antibacterial and antiviral properties . Moreover, a compounding strategy was applied to make the fibers with simultaneously high antibacterial and antiviral activity , especially in short contact time. The bacteriostatic rate (against S. aureus : 95.81 %, against E. coli : 92.07 %, 1 h) of the compounded fibers improved substantially when a few GCOS-CFs were mixed with SCOS-CFs; especially, it was much higher than both the individual GCOS-CFs and SCOS-CFs. By contrast, the improvement of the antiviral properties was less dramatic; however, even a few SCOS-CFs was mixed, the antiviral properties increased pronouncedly. Although the electrostatic interaction between SCOS and GCOS can make the SCOS-GCOS mixture lose some extent of antibacterial activity , the long chains of cellulose restrain the electrostatic interaction between sulfonic and amino groups, leading to their synergistic action and eventually superior antibacterial and antiviral effects.
研究领域:
材料科学
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生物技术
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纺织工程
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DOI
PubMed
高影响因子
CARBOHYDRATE POLYMERS
·
2017
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IF 4.811
Lin Yue,Jingru Li,Wanwen Chen,Xiaoli Liu,Qixing Jiang,Wenshui Xia
The novel derivatives of geraniol grafted chitosan oligosaccharide were synthesized via substitution and deprotection reaction, respectively. The products were identified by Fourier transform infrared (FT-IR), 1 H nuclear magnetic resonance (NMR), X-ray diffraction analysis (XRD), Thermogravimetric analysis (TGA) and UV–vis absorption spectroscopy. It is revealed that the derivatives exhibited a good solubility, thermal stability and antibacterial properties.
研究领域:
生物材料学
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微生物学
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产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
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2018
·
IF 4.946
Zhen Qin,Qiming Chen,Si Lin,Sa Luo,Yongjun Qiu,Liming Zhao
Chitooligosaccharide is widely used as a functional food additive and a valuable pharmacological agent. The transformation of chitinous biomass into valuable bioactive chitooligosaccharides is one of the most exciting applications of chitosanase. A novel glycoside hydrolase (GH) family 46 chitosanase (GsCsn46A) from rhizobacterium Gynuella sunshinyii was cloned and heterologously expressed in Escherichia coli . GsCsn46A showed maximal activity at pH 5.5 and 30 °C. GsCsn46A featured remarkable cold-adapted property, which controllably hydrolyzed chitosan to three types of chitooligosaccharides at the mild reaction condition (reaction condition: pH 5.5 at 30 °C; method for stopping the reaction: 50 °C for 30 min). The yields of three types of chitooligosaccharides products (degree of polymerization (DP): 2–7, 2–5 and 2–3) were 70.9%, 87.1% and 94.6% respectively. This novel cold-adapted chitosanase provides a cleaner production process for the controllable preparation of chitooligosaccharides with the specific DP.
研究领域:
生物化学
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酶工程
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食品科学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2019
·
IF 3.571
Xingchen Zhai,Shoujun Yuan,Xin Yang,Pan Zou,Linna Li,Guoyou Li,Yong Shao,A. M. Abd El-Aty,Ahmet Hacımüftüoğlu,Jing Wang
In recent years, various studies have confirmed the role of natural products as effective cancer prevention and treatment drugs. The present study demonstrated that chitosan oligosaccharide (COS) from shells of shrimp and crab caused an inhibitory effect on the proliferation of human renal carcinoma in vitro and in vivo. First, the in vivo biodistribution of COS was investigated by the synthesis of cyanine-7-labeled COS (COS–Cy7) following tail vein injection. The kidney was found to be a major target organ. Then, the impacts on renal carcinoma cell proliferation, apoptosis, and reactive oxygen species (ROS) production were observed in vitro, and an orthotopic xenograft tumor model was designed to evaluate the antitumor efficacy of COS in vivo. In renal carcinoma cells, COS induced G2/M phase arrest and apoptosis in a ROS-dependent fashion. COS significantly promoted mRNA expression of nuclear factor erythroid 2-related factor (Nrf2) and Nrf2 target genes, such as heme oxygenase 1, modifier subunit of glutamate cysteine ligase, and solute carrier family 7 member 11. Additionally, COS significantly upregulated the protein expression of glucose-regulated protein 78, protein RNA-like endoplasmic reticulum (ER) kinase, eukaryotic initiation factor 2α, activating transcription factor 4, C/EBP homologous protein, and cytochrome c, which justified the activation of the ER stress signaling pathway. In vivo, COS repressed tumor growth and induced apoptosis and ROS accumulation, consistent with the in vitro results. Taken together, COS repressed human renal carcinoma growth and induced apoptosis both in vitro and in vivo, mainly via ROS-dependent ER stress pathways.
研究领域:
肿瘤学
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药理学
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生物化学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2018
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IF 3.909
Qian Zhou,Li Cui,Lamei Ren,Ping Wang,Chao Deng,Qiang Wang,Xuedong Fan
Chitooligosaccharide (COS) has the characteristic of antioxidant and antibacterial effects. To improve the properties of silk fibroin (SF), COS was enzymatically grafted onto SF membranes using a laccase from Trametes versicolor . d -Glucosamine hydrochloride (GAH) and p -hydroxyphenylacetamide (PHAD), as the model compound of COS and tyrosine residues in SF were utilized to disclose the grafting mechanism, respectively. The data from UPLC-TQD and GPC analysis implied that laccase might catalyze the oxidation of PHAD and led to the formation of self-polymerized products. FTIR and 1 H NMR results verified the occurrence of the laccase-assisted reactions between COS and PHAD. For the fibroin samples with different treatments, incubation with laccase alone led to remarkable increase in the molecular weight of SF, mainly owing to the efficient self-crosslinks of the fibroin chains. For the COS-grafted SF membrane, there was no obvious change in the thermal behavior, while the antioxidant and antibacterial properties were evidently improved when compared to that of the untreated. Meanwhile, biocompatibility of the COS grafted SF membrane was acceptable according to the cell viability of NIH/3T3 cells. The present work provides a novel method for preparation of the multifunctional fibroin-based biomaterials.
研究领域:
生物材料学
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酶工程
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医学材料
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产品关联
DOI
PubMed
高影响因子
Frontiers in Pharmacology
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2018
·
IF 3.831
Guo Xi,Sun Tong,Zhong Rui,Ma Lu,You Chao,Tian Meng,Li Hao,Wang Chengwei
Chitosan oligosaccharide (COS) is known for its unique biological activities such as anti-tumor, anti-inflammatory, anti-oxidant, anti-bacterial activity, biological recognition, and immune enhancing effects, and thus continuous attracting many research interests in drug, food, cosmetics, biomaterials and tissue engineering fields. In comparison to its corresponding polymer, COS has much higher absorption profiles at the intestinal level, which results in permitting its quick access to the blood flow and potential contacting with blood components. However, the effects of COS on blood components remain unclear to date. Herein, two COS with different molecular weight (MW) were characterized by FTIR and 1H NMR, and then their effects on human blood components, including red blood cells (RBCs) (hemolysis, deformability, and aggregation), coagulation system [activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), and the concentration of fibrinogen (Fib)], complement (C3a and C5a activation), and platelet (activation and aggregation), were comprehensively studied. In the case of RBCs, COS exhibited a low risk of hemolysis in a dose and molecular weight dependent manner and the irreversible aggregation was observed in their high concentration. For coagulation system, COS has a mild anticoagulation activity through blocking the intrinsic coagulation pathway. In addition, COS showed no effect on complement activation in C3a and C5a and on platelet activation while inhibition of platelet aggregation was evident. Finally, the mechanism that effects of COS on blood components was discussed and proposed.
研究领域:
医学
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生物化学
·
药理学
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产品关联
DOI
PubMed
高影响因子
ARABIAN JOURNAL FOR SCIENCE AND ENGINEERING
·
2019
·Vol. 45
·期 1
·29-40
·
IF 1.518
Liu Xiaoli,Xie Wancui,Yang Xihong,Zhan Xiaobei,Xia Wenshui
The aim of this work was to develop a novel, nontoxic, bioactive antimicrobial material based on a natural polymer, chitosan oligosaccharides (COS), a natural pyrone compound, kojic acid (KA), and piperazine. The COS-based polymer (COS-O-MB) has been synthesized by selective partial alkylation reaction to graft kojic acid, KA, and piperazine onto the COS molecular chain. Then, physicochemical properties and bioactivity of the COS-O-MBs were evaluated and it was characterized by FT-IR, NMR spectroscopy, Mw, PID, antimicrobial activity, hemolysis assay, biocompatibility, animal toxicity, and antimicrobial action mode studies. The results revealed that the alkylation reaction took place at the C-6 position of COS, and the as-prepared COS-O-MB significantly enhanced the antimicrobial activity and biocompatibility and reduced the hemolytic activity in vitro in comparison with that of the free COS and KA. Meanwhile, the antimicrobial action mode studies proved that COS-O-MB can pass through the anionic microbial membrane, destroy the cytomembrane integrity, and potentially affect protein denaturation, demonstrating that exposure to COS-O-MB results in the death of microbial cells. But importantly, COS-O-MB is not toxic to human cells. Hence, low hemolytic activity to human red blood cells, and nontoxic nature to female mice of SLAC KM strain, made this novel polymer material a promising and effective compound for food and pharmaceutical industries.
研究领域:
生物材料学
·
微生物学
·
药理学
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产品关联
DOI
高影响因子
FOOD CHEMISTRY
·
2019
·
IF 5.399
Lin Yue,Dan Sun,Imran Mahmood Khan,Xiaoli Liu,Qixing Jiang,Wenshui Xia
The present study aims at synthesizing and in vitro antibacterial activity evaluation of chitosan oligosaccharide (COS) modified by Cinnamyl alcohol (Cin) onto the OH position of COS . Three different degrees of substitution (DS) COS-O-Cin1-3 were synthesized by changing different molar ratios of COS to Cin. UV–visible spectroscopy (UV–vis), Fourier transform infrared (FT-IR), 1 H nuclear magnetic resonance ( 1 H NMR), thermogravimetric analysis (TGA), X-ray diffraction (XRD) and elemental analysis were conducted to characterize the successful synthesis of COS-O-Cin1-3. The results showed that they exhibited higher thermal stability, weaker crystallinity and better antibacterial properties than that of COS . These results aided in obtaining the important supports for exploring new functional antibacterial agents, which expand the scope of COS’s application in the food industry.
研究领域:
食品科学
·
生物化学
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DOI
高影响因子
ENERGY & FUELS
·
2024
·
IF 5.2
Jie Yu,Hongping Quan,Jia Xiao,Jingran Wang,Lilong Zhang,Xueshuo Chang,Shuaibin Peng,Jinyu Wang,Zhiyu Huang
Based on dispersing the stacking structure of asphaltenes and inhibiting the reaggregation of asphaltenes, functional chitosan oligosaccharide (COS) derivatives were prepared using COS in this study. Driven by the conjugated intercalation of aromatic rings, COS derivatives could move among the lamellae of asphaltene stacked structures. The effect of COS derivatives on asphaltene aggregates for face-to-face stacking and the inner diameter of the aromatic sheet was characterized by X-ray diffraction and Raman. The microstructure of asphaltene in this mode of interaction could be observed by field emission scanning electron microscopy and high-resolution transmission electron microscopy. The effect of COS derivatives on the stacking structure and aggregation behavior of asphaltenes could enhance the fluidity of heavy oil, resulting in a change in the viscosity-temperature curve of heavy oil. This demonstrates the enormous research value of COS derivatives in the extraction and transportation of heavy oil.
研究领域:
石油化学
·
材料科学
·
生物学
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产品关联
DOI
高影响因子
Polymers
·
2021
·Vol. 13
·期 17
·2929
·
IF 4.329
Jing Miao,Liwen Zhang,Peng Gao,Huawei Zhao,Xianji Xie,Junyan Wang
Glucose Transporter-1 (GLUT-1) is considered to be a possible intrinsic marker of hypoxia in malignant tumors, which is an important factor in radioresistance of laryngocarcinoma. We speculated that the inhibition of GLUT-1 expression might improve the radiosensitivity of laryngocarcinoma. GLUT-1 siRNA was designed to inhibit the GLUT-1 expression, but the high molecular weight and difficult drug delivery limited the application. Herein, we constructed a glycolipid polymer chitosan oligosaccharide grafted stearic acid (CSSA) to conjugate siRNA via electrostatic interaction. The characteristics of CSSA and CSSA/siRNA were studied, as well as the radiosensitization effect of siRNA on human laryngocarcinoma epithelial (Hep-2) cells. Compared with the traditional commercial vector LipofectamineTM2000 (Lipo), CSSA exhibited lower cytotoxicity, more efficiently cellular uptake. Incubating with CSSA/siRNA, the survival rates of Hep-2 cells were significantly decreased comparing with either the group before transfection or Lipo/siRNA. CSSA is a promising carrier for efficient siRNA delivery and radiosensitization of laryngocarcinoma.Keywords:GLUT-1;siRNA;chitosan oligosaccharide grafted stearic acid;laryngocarcinoma;radiosensitization
研究领域:
癌症生物学
·
药学
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
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