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superoxide dismutase genes in Pyropia haitanensis:molecular cloning,characterization and mRNA expression

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Acta Oceanologica Sinica 2016年第6期35卷 101-111页

作者： JI Dehua XU Yan XIAO Haidong CHEN Changsheng XU Kai XIE Chaotian Fisheries College Jimei University

superoxide dismutase（SOD,EC 1.***.1.1） is the first and most important line of cellular defense against oxidative stress.In this study,based on unigene sequences of P.haitanensis,three full-length Ph SOD genes were obtained by RACE technology,and named Ph MSD,Ph CSD1 and Ph CSD2.The full-length c DNAs of these genes comprised973,1 029 and 954 nucleotides,respectively.The c DNAs encoded proteins of 224,134 and 216 amino acids,with isoelectric points of 5.75,4.65 and 10.74,respectively.Based on their conserved motifs and phylogenetic tree analysis,the three Ph SODs were divided into two SOD types：Ph MSD is a Mn-SOD,and Ph CSD1 and Ph CSD2 are Cu Zn-SODs.q PCR was used to measure the expression of the three Ph SOD genes in different life phases of P.haitanensis,and under different periods of high-temperature stress and different levels of desiccation.In the different life phases of P.haitanensis,the expression levels of the three Ph SODs were all significantly higher in the thallus than in the conchocelis.During high-temperature and desiccation stress,the expression levels of Ph CSD1 and Ph CSD2 were highly induced by the O2-content,but the expression level of Ph MSD was highly depressed by high temperature stress and showed no significant change during desiccation.

关键词： superoxide dismutase gene cloning desiccation stress high-temperature stress qPCR

superoxide dismutase mimetic ability of Mn-doped ZnS QDs

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Chinese Chemical Letters 2018年第12期29卷 1865-1868页

作者： Xiangling Ren Mingqian Wang Xing He Zheng Li Jing Zhang Wei Zhang Xudong Chen Hong Ren Xianwei Meng Laboratory of Controllable Preparation and Application of Nanomaterials Key Laboratory of CryogenicsTechnical Institute of Physics and ChemistryChinese Academy of SciencesBeijing 100190China First Affiliated Hospital of Southern University of Science and Technology Shenzhen People's Hospital&Shenzhen Interventional Medical Engineering Research and Development CenterShenzhen 518020China Beijing University of Chemical Technology Beijing 100029China Beijing Technology&Business University Beijing Higher Institution Engineering Research Center of Food Additives and IngredientsBeijing Key Laboratory of Flavor ChemistryBeijing Laboratory for Food Quality and SafetyBeijing 100048China University of Chinese Academy of Sciences Beijing 100049China

superoxide dismutase(SOD) is an important antioxidant enzyme in the body. SOD has special physiological activity and is the primary substance for scavenging free radicals in living organisms.However, the expensive and complex extraction processes, low SOD yield, as well as difficult to store at room temperature have seriously hindered its application pace. Herein, the enzyme mimetic function of Mn-doped ZnS quantum dots(QDs) was discovered. The improved Marklund and McCord method both showed that Mn-doped ZnS QDs possess intrinsic SOD-like activity. The effects of temperature and pH on the mimetic enzyme activity of Mn-doped ZnS QDs have been investigated compared with SOD enzymes.The low cost and easy to synthesize white Mn-doped ZnS QDs with good biocompatibility are expected to be used as a new type of SOD nanozymes in the biology-relevant fields.

关键词： Mn-doped ZnS QDs superoxide dismutase Nanozymes Marklund method Pyrogallol

Vacancies-rich CoAl monolayer layered double hydroxide as efficient superoxide dismutase-like nanozyme

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Nano Research 2022年第9期15卷 7940-7950页

作者： Shuo Zhang Jun Chen Wen-Sheng Yang Xu Chen State Key Laboratory of Chemical Resource Engineering Beijing University of Chemical TechnologyBeijing 100029China

With the extensive development of nanozymes in recent years,catalytic efficiency has been considered as the Gordian knot that restricts the further applications of nanozyme.Moreover,the usage of layered double hydroxides(LDHs)as antioxidant nanozymes for scavenging reactive oxygen species(ROS)has not been studied.Herein,we report a vacancies-rich monolayer cobalt-alumina LDH nanosheet(m-CoAl-LDH)through a facile direct synthesis method as an efficient nanozyme with superoxide dismutase(SOD)-like activity.The m-CoAl-LDH exhibits a record-breaking catalytic efficiency with the catalytic constant(Kcat)as high as 4.33×10^(11)M^(−1)·s−1.Elucidated by the experimental and theoretical studies,the abundant oxygen and metal vacancies were existed in the single layer for the outstanding performance owing to the increased active sites.In addition,density functional theory(DFT)calculations further reveal the significant role of synergistic effect of oxygen and metal vacancies in reducing the adsorption energy of superoxide(O_(2)^(•−)),which improves the catalytic performance.The m-CoAl-LDH nanosheets were applied in cells to relieving the oxidative damage caused by O_(2)^(•−)in mitochondria.Such vacancy-rich monolayer CoAl-LDH nanosheets represent the first example of LDH nanozyme with specific SOD-like activity as well as record high catalytic efficiency,which may provide deeper insight into efficient nanozyme design by defect-engineering.

关键词： nanozymes superoxide dismutase layered double hydroxides defect catalytic efficiency

Oral delivery of superoxide dismutase by lipid polymer hybrid nanoparticles for the treatment of ulcerative colitis

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Chinese Chemical Letters 2022年第10期33卷 4617-4622页

作者： Yaxin Cui Tianyu Zhu Xueyan Zhang Jicong Chen Fengying Sun Youxin Li Lesheng Teng School of Life Sciences Jilin UniversityChangchun 130012China

Protein-based drugs have received extensive attention in the field of drug research in recent years.However,protein-based drug activity is difficult to maintain during oral delivery,which limits its application.This study developed bifunctional oral lipid polymer hybrid nanoparticles(R8-PEG-PPNPs)that deliver superoxide dismutase(SOD)for the treatment of ulcerative colitis(UC).R8-PEG-PPNPs was composed of PCADK,PLGA,lecithin,and co-modified with stearic acid-octa-arginine and polyethylene glycol.The nanoparticles(NPs)are uniformly dispersed with a complete spherical structure.In vitro stability and release studies showed that R8-PEG-PPNPs exhibited good stability and protection.In vitro cell culture experiments demonstrated that R8-PEG-PPNPs as carriers have no significant toxic effects on cells at concentration below 1000µg/mL and promote cellular uptake.In experiments with ulcerative colitis mice,R8-PEG-PPNPs were able to enhance drug absorption by intestinal epithelial cells and accumulate effectively at the site of inflammation.Its therapeutic effect further demonstrates that R8-PEG-PPNPs are a promising delivery system for oral delivery of protein-based drugs.

关键词： superoxide dismutase PCADK DSPE-PEG Cell-penetrating peptide Ulcerative colitis Lipid polymer hybrid nanoparticles

Oxygen vacancy enhanced biomimetic superoxide dismutase activity of CeO_(2)-Gd nanozymes

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Journal of Rare Earths 2021年第9期39卷 1108-1116页

作者： Xiangcheng Shi Jingjie Yang Xintong Wen Fuii Tian Changyan Li College of Chemistry and Chemical Engineering Inner Mongolia UniversityHohhot010021China Department of Chemistry National University of Singapore.Singapore 117543Singapore Key Laboratory of Medicinal and Edible Plants Resources of Hainan Province Hainan Vocational University of Science and TechnologyHaikou 571126China

Cerium oxide-based nanozymes have recently drawn much attention in the field of biomedical and antioxidant applications,because of their unique regenerative or autocatalytic properties.Herein,we studied the biomimetic superoxide dismutase(SOD)nanozymes CeO_(2)-Gd that combines the fluorescence properties of rare earth Gd with the antioxidant properties of CeO_(2)nanoparticles,which was prepared via facile route.The structure and composition of the CeO_(2)-Gd were measured and verified by X-ray powder diffraction(XRD),Raman spectroscopy,transmission electron microscopy(TEM),energydispersive X-ray spectroscopy(EDX),and X-ray electron spectroscopy(XPS).Confocal microscopy was used to image cells.Antioxidant performance and cell viability of these nanozymes were measured in vitro using BGC-803 cell.CeO_(2)-Gd nanozymes with a higher Ce^(3+)/Ce^(4+)ratio show higher superoxide dismutase(SOD)mimetic activity.Their antioxidant activity and fluorescence properties of CeO_(2)-Gd in BGC-803 cancer cells are enhanced by oxygen vacancies generated by doping rare-earth elements Gd.This work may guide the future design of CeO_(2)-Gd-based biomimetic nanozymes for anticancer and antioxidant applications.

关键词： Antioxidant enzyme CeO_(2)-Gd nanozymes superoxide dismutase Biomimetic nanozymes Oxygen vacancy Rare earths

Extracellular superoxide dismutase VdSOD5 is required for virulence in Verticillium dahliae

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Journal of Integrative Agriculture 2021年第7期20卷 1858-1870页

作者： TIAN Li HUANG Cai-min ZHANG Dan-dan LI Ran CHEN Jie-yin SUN Wei-xia QIU Nian-wei DAI Xiao-feng College of Life Science Qufu Normal UniversityQufu 273165P.R.China State Key Laboratory for Biology of Plant Diseases and Insect Pests/lnstitute of Plant Protection Chinese Academy of Agricultural SciencesBeijing 100193P.R.China

Plants produce reactive oxygen species(ROS) to defend pathogens. To counteract this attack, certain pathogens express superoxide dismutases(SODs) to scavenge host-derived ROS. However, the roles of SODs in Verticillium dahliae, an important vascular pathogen, are not clear. Our previous study has shown that a putative extracellular SOD(VdSOD5) of V. dahliae is significantly induced by culturing in cotton tissues, suggesting that VdSOD5 may play an important role in host–pathogen interactions and virulence. Here, we showed that VdSOD5 encoded a superoxide dismutase with a cofactor copper-binding site and a functional signal peptide that can conduct protein secretion in an invertase-mutated yeast strain. The mutations in VdSOD5(ΔVdSOD5) did not change the normal vegetative growth and conidial production but reduced the virulence of V. dahliae on susceptible host cotton. Further studies showed that the transcription of Vd SOD5 was significantly up-regulated during the early stage of infection, and the loss-of-function of VdSOD5 decreased culture filtrate and fungal tissue SOD activities of V. dahliae by 74 and 28%, respectively. Compared to the wild-type strain Vd991, the ΔVdSOD5 showed the same sensitivity to the intracellular ROS generator menadione. Furthermore, nitroblue tetrazolium(NBT) staining demonstrated that VdSOD5 functioned in the detoxification of superoxides generated by host roots during infection. These results suggest that VdSOD5 of V. dahliae is an important virulence factor, secreted out of cells to combat host-derived ROS.

关键词： Verticillium dahliae superoxide dismutase secretion virulence ROS detoxification

Pulmonary endothelium-targeted nanoassembly of indomethacin and superoxide dismutase relieves lung inflammation

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Acta Pharmaceutica Sinica B 2023年第11期13卷 4607-4620页

作者： Yi Yang Makhloufi Zoulikha Qingqing Xiao Feifei Huang Qi Jiang Xiaotong Li Zhenfeng Wu Wei He Shanghai Skin Disease Hospital Tongji University School of MedicineShanghai 200443China School of Pharmacy China Pharmaceutical UniversityNanjing 2111198China Department of Traditional Chinese Medicine Processing and Preparation Nanjing University of Chinese MedicineNanjing 210023China Key Laboratory of Modern Preparation of TCM Ministry of EducationJiangxi University of Chinese MedicineNanchang 330004China

Lung inflammation is an essential inducer of various diseases and is closely related to pulmonary-endothelium dysfunction.Herein,we propose a pulmonary endothelium-targeted codelivery system of anti-inflammatory indomethacin(IND)and antioxidant superoxide dismutase(SOD)by assembling the biopharmaceutical SOD onto the“vector”of rod-like pure IND crystals,followed by coating with anti-ICAM-1 antibody(Ab)for targeting endothelial cells.The codelivery system has a 237 nm diameter in length and extremely high drug loading of 39%IND and 2.3%SOD.Pharmacokinetics and biodistribution studies demonstrate the extended blood circulation and the strong pulmonary accumulation of the system after intravenous injection in the lipopolysaccharide(LPS)-induced inflammatory murine model.Particularly,the system allows a robust capacity to target pulmonary endothelium mostly due to the rod-shape and Ab coating effect.In vitro,the preparation shows the synergistic antiinflammatory and antioxidant effects in LPS-activated endothelial cells.In vivo,the preparation exhibits superior pharmacodynamic efficacy revealed by significantly downregulating the inflammatory/oxidative stress markers,such as TNF-a,IL-6,COX-2,and reactive oxygen species(ROS),in the lungs.In conclusion,the codelivery system based on rod-like pure crystals could well target the pulmonary endothelium and effectively alleviate lung inflammation.The study offers a promising approach to combat pulmonary endothelium-associated diseases.

关键词： Codelivery Nanocrystals superoxide dismutase Indomethacin Pulmonary endothelium Inflammation Acute lung injury

Cloning of a putative extracellular Cu/Zn superoxide dismutase and functional differences of superoxide dismutases in invasive and indigenous whiteflies

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Insect Science 2015年第1期22卷 52-64页

作者： Xian-Long Gao Jun-Min Li Hong-Xing Xu Gen-Hong Yan Min Jiu Shu-Sheng Liu Xiao-Wei Wang Ministry of Agriculture Key Laboratory of Agricultural Entomology Institute of Insect Sciences Zhejiang University Hangzhou Instituteof Virology and Biotechnology Zhejiang Academy of Agricultural Sciences Hangzhou College of Food and Bioengineering HenanUniversity of Science and Technology Luoyang China

superoxide dismutases （SODs） are a group of important antioxidant defense enzymes. In this study, a putative extracellular Cu/Zn superoxide dismutase （ecCuZnSOD） complementary DNA was cloned and characterized from the whitefly, Bemisia tabaci. Quantitative polymerase chain reaction analysis showed that the expression level of BtecCuZnSOD was more than 10-fold higher in the invasive Middle East Asia Minor 1 （MEAM1） than in the native Asia II 3 species of the B. tabaci species complex. After exposure to low temperature （4 ℃）, the expression of Bt-ecCuZnSOD gene was significantly up-regulated in MEAM1 but not in Asia II 3. Furthermore, the expression level ofB. tabaci intracellular CuZnSOD （Bt-icCuZnSOD）, Bt-ecCuZnSOD and mitochondrial MnSOD （Bt-mMnSOD） was compared after transferring MEAM1 and Asia II 3 whiteflies from favorable （cotton） to unfavorable host plants （tobacco）. On cotton, both CuZnSOD genes were expressed at a higher level in MEAM1 compared with Asia II 3. Interestingly, after transferring onto tobacco, the expression of Bt-ecCuZnSOD was significantly induced in Asia II 3 but not in MEAM1. On the other hand, while Bt-mMnSOD was expressed equally in both species on cotton, Bt-mMnSOD messenger RNA was up-regulated in MEAM 1 on tobacco. Consistently, enzymatic activity assays of CuZnSOD and MnSOD demonstrated that CuZnSOD might play an important protective role against oxidative stress in Asia II 3, whereas MnSOD activation was critical for MEAM1 whiteflies during host adaptation. Taken together, our results suggest that the successful invasion ofMEAM 1 is correlated with its constitutive high activity of CuZnSOD and inducible expression of MnSOD under stress conditions.

关键词： Bemisia tabaci extracellular Cu/Zn superoxide dismutase oxidative strees superoxide dismutase whitefly

Purification and biochemical characterization of manganesecontaining superoxide dismutase from deep-sea thermophile Geobacillus sp. EPT3

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Acta Oceanologica Sinica 2014年第12期33卷 163-169页

作者： ZHU Yanbing LI Hebin NI Hui LIU Jingwen XIAO Anfeng CAI Huinong College of Food and Biological Engineering Jimei University Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering Research Center of Food Microbiology and Enzyme Engineering Technology of Fujian Province Xiamen Medical College

Thermostable SOD is a promising enzyme in biotechnological applications. In the present study, thermo-phileGeobacillussp.EPT3 was isolated from a deep-sea hydrothermal field in the East Pacific. A thermo-stable superoxide dismutase （SOD） from this strain was purified to homogeneity by steps of fractional am-monium sulfate precipitation, DEAE-Sepharose chromatography, and Phenyl-Sepharose chromatography. SOD was purified 13.4 fold to homogeneity with a specific activity of 3 354 U/mg and 11.1% recovery. SOD fromGeobacillussp. EPT3 was of the Mn-SOD type, judged by the insensitivity of the enzyme to both KCN and H2O2. SOD was determined to be a homodimer with monomeric molecular mass of 26.0 kDa. It had high thermostability at 50＆#176;C and 60＆#176;C. At tested conditions,SOD was relatively stable in the presence of some inhibitors and denaturants, such asβ-mercaptoethanol （β-ME）, dithiothreitol （DTT）, phenylmethylsulfonyl fluoride （PMSF）, urea, and guanidine hydrochloride.Geobacillussp. EPT3 SOD showed striking stability across a wide pH range from 5.0 to 11.0. It could withstand denaturants of extremely acidic and alkaline conditions, which makes it useful in the industrial applications.

关键词： Geobacillus sp. superoxide dismutase thermostability purification