Pretreated wheat insoluble arabinoxylan was converted to oligosaccharides of structural variants using combinatorial enzyme approach. The digestive products were separated by preparative scale chromatographic Amberlit...Pretreated wheat insoluble arabinoxylan was converted to oligosaccharides of structural variants using combinatorial enzyme approach. The digestive products were separated by preparative scale chromatographic Amberlite XAD-2 column. Fractions containing feruloyl oligosaccharides (FOS) were isolated, pooled, freeze-dried, and demonstrated to possess antimicrobial activity. The FOS suppressed cell growth of the test organism ATCC 8739 E. coli with a MIC value of 0.028% (w/v, 35˚C, 24 hr). The antimicrobial action was observed exceeding 72 hr of culture incubation. The FOS product could be a useful source of prebiotics or preservatives. The present results further confirm the science and application of the concept of combinatorial enzyme technique.展开更多
Unlike chemosynthetic drugs designed for specific molecular and disease targets,active small-molecule natural products typically have a wide range of bioactivities and multiple targets,necessitating extensive screenin...Unlike chemosynthetic drugs designed for specific molecular and disease targets,active small-molecule natural products typically have a wide range of bioactivities and multiple targets,necessitating extensive screening and development.To address this issue,we propose a strategy for the direct in situ microdynamic examination of potential drug candidates to rapidly identify their effects and mechanisms of action.As a proof-of-concept,we investigated the behavior of mussel oligosaccharide(MOS-1)by tracking the subcellular dynamics of fluorescently labeled MOS-1 in cultured cells.We recorded the entire dynamic process of the localization of fluorescein isothiocyanate(FITC)-MOS-1 to the lysosomes and visualized the distribution of the drug within the cell.Remarkably,lysosomes containing FITC-MOS-1 actively recruited lipid droplets,leading to fusion events and increased cellular lipid consumption.These drug behaviors confirmed MOS-1 is a candidate for the treatment of lipid-related diseases.Furthermore,in a high-fat HepG2 cell model and in high-fat diet-fed apolipoprotein E(ApoE)^(-/-)mice,MOS-1 significantly promoted triglyceride degradation,reduced lipid droplet accumulation,lowered serum triglyceride levels,and mitigated liver damage and steatosis.Overall,our work supports the prioritization of in situ visual monitoring of drug location and distribution in subcellular compartments during the drug development phase,as this methodology contributes to the rapid identification of drug indications.Collectively,this methodology is significant for the screening and development of selective small-molecule drugs,and is expected to expedite the identification of candidate molecules with medicinal effects.展开更多
Alginate oligosaccharides(AOS)and fucoidan are two important bioactive carbohydrate which that can be prepared from brown seaweeds.We investigated a novel enzyme-assisted one-pot method on AOS and fucoidan extraction ...Alginate oligosaccharides(AOS)and fucoidan are two important bioactive carbohydrate which that can be prepared from brown seaweeds.We investigated a novel enzyme-assisted one-pot method on AOS and fucoidan extraction from Lessonia trabeculata,and found that the optimum extraction procedure was:first,using 10-g/L Na_2CO_3 solution to digest the seaweeds for 2 h at 80℃ with the material-liquid ratio 1?20(g/mL),and then adding 4.0%fucoidan cleavage enzyme for reaction for 12 h at 45℃.The yield of AOS and fucoidan was 21.36%and 14.40%,respectively,which was higher than those using traditional methods.The degree of polymerization of AOS was determined to be 3-5,confirming the results through thin layer chromatography(TLC)and electrospray ionization mass spectrometry(ESI-MS).The ratio of guluronic acid to mannuronic acid in AOS was found to be 1.97.Through anion-exchange column chromatography,four sulfated polysaccharide fractions(F0.5,F1.0,F1.5,and F2.0)were successfully isolated from fucoidan.Chemical analysis revealed that fucoidan and F0.5 were heteropolysaccharides,consisting mainly of mannuronic acid,fucose,and xylose,while the other fractions were predominantly composed of fucose and galactose.The sulfate group content increased from 8.74%to 18.76%,while the uronic acid content decreased from 23.79%to 5.58%across the fractions.All fractions of fucoidan exhibited significant antioxidant activity in various tests.There was a positive correlation between the sulfate content and the ability to scavenge superoxide radicals.Additionally,both AOS and the fucoidan fractions showed immunostimulation activity and were nontoxic to RAW264.7 cells.These results indicate that the novel enzyme-assisted one-pot method is a green and efficient approach for the extraction of fucoidan and alginate oligosaccharides from brown seaweeds.It has potential applications in the processing industry of brown seaweeds.展开更多
Alginate oligosaccharides(AOS)enhance drought resistance in wheat(Triticum aestivum L.),but the definite mechanisms remain largely unknown.The physiological and transcriptome responses of wheat seedlings treated with ...Alginate oligosaccharides(AOS)enhance drought resistance in wheat(Triticum aestivum L.),but the definite mechanisms remain largely unknown.The physiological and transcriptome responses of wheat seedlings treated with AOS were analyzed under drought stress simulated with polyethylene glycol-6000.The results showed that AOS promoted the growth of wheat seedlings and reduced oxidative damage by improving peroxidase and superoxide dismutase activities under drought stress.A total of 10,064 and 15,208 differentially expressed unigenes(DEGs)obtained from the AOS treatment and control samples at 24 and 72 h after dehydration,respectively,were mainly enriched in the biosynthesis of secondary metabolites(phenylpropanoid biosynthesis,flavonoid biosynthesis),carbohydrate metabolism(starch and sucrose metabolism,carbon fixation in photosynthetic organisms),lipid metabolism(fatty acid elongation,biosynthesis of unsaturated fatty acids,alpha-linolenic acid metabolism,cutin,suberine and wax biosynthesis),and signaling transduction pathways.The up-regulated genes were related to,for example,chlorophyll a-b binding protein,amylosynthease,phosphotransferase,peroxidase,phenylalanine ammonia lyase,flavone synthase,glutathione synthetase.Signaling molecules(including MAPK,plant hormones,H_(2)O_(2) and calcium)and transcription factors(mainly including NAC,MYB,MYB-related,WRKY,bZIP family members)were involved in the AOS-induced wheat drought resistance.The results obtained in this study help underpin the mechanisms of wheat drought resistance improved by AOS,and provides a theoretical basis for the application of AOS as an environmentally sustainable biological method to improve drought resistance in agriculture.展开更多
利用实际航摄资料评价了WGS 84和国家80坐标系下机载POS(position and orientation system)系统直接对地目标定位的精度,分析了不同大地水准面拟合方法对高程精度的影响,检校了POS系统的视准轴误差,讨论了检校场的检校结果。试验表明,基...利用实际航摄资料评价了WGS 84和国家80坐标系下机载POS(position and orientation system)系统直接对地目标定位的精度,分析了不同大地水准面拟合方法对高程精度的影响,检校了POS系统的视准轴误差,讨论了检校场的检校结果。试验表明,基于POS系统的航空遥感直接对地目标定位至少需要利用带有1个平高地面控制点的检校场对POS系统进行检校才能消除系统误差;在WGS84坐标系中,可以获得较高的定位精度,而转换到国家80坐标系下,必须对高程进行大地水准面拟合改正。展开更多
文摘Pretreated wheat insoluble arabinoxylan was converted to oligosaccharides of structural variants using combinatorial enzyme approach. The digestive products were separated by preparative scale chromatographic Amberlite XAD-2 column. Fractions containing feruloyl oligosaccharides (FOS) were isolated, pooled, freeze-dried, and demonstrated to possess antimicrobial activity. The FOS suppressed cell growth of the test organism ATCC 8739 E. coli with a MIC value of 0.028% (w/v, 35˚C, 24 hr). The antimicrobial action was observed exceeding 72 hr of culture incubation. The FOS product could be a useful source of prebiotics or preservatives. The present results further confirm the science and application of the concept of combinatorial enzyme technique.
基金supported by Shandong Province Key R&D Program,China(Major Technological Innovation Project)(Grant No.:2021CXGC010501)Young Elite Scientists Sponsorship Program by China Association of Chinese Medicine,China(Grant No.:CACM-2023-QNRC1-02)+8 种基金the National Natural Science Foundation of China(Grant Nos.:22107059,22007060,82302743)the Natural Science Foundation of Shandong Province,China(Grant Nos.:ZR2022QH304,ZR2021QH057,ZR2020QB166)the Program for Youth Innovation Technology in Colleges and Universities of Shandong Province of China(Grant No.:2021KJ035)Taishan Scholars Program,China(Grant Nos.:TSQN202211221,TSPD20181218)Shandong Science Fund for Excellent Young Scholars,China(Grant No.:ZR2022YQ66)Shandong Province Traditional Chinese Medicine Science and Technology Project,China(Grant No.:Q-2023059)Shenzhen Basic Research Project,China(Grant No.:JCYJ20190809160209449)the General Project of Shandong Natural Science Foundation,China(Grant No.:ZR2021MH341)Jinan Innovation Team Project of Colleges and Universities,China(Grant No.:2021GXRC072).
文摘Unlike chemosynthetic drugs designed for specific molecular and disease targets,active small-molecule natural products typically have a wide range of bioactivities and multiple targets,necessitating extensive screening and development.To address this issue,we propose a strategy for the direct in situ microdynamic examination of potential drug candidates to rapidly identify their effects and mechanisms of action.As a proof-of-concept,we investigated the behavior of mussel oligosaccharide(MOS-1)by tracking the subcellular dynamics of fluorescently labeled MOS-1 in cultured cells.We recorded the entire dynamic process of the localization of fluorescein isothiocyanate(FITC)-MOS-1 to the lysosomes and visualized the distribution of the drug within the cell.Remarkably,lysosomes containing FITC-MOS-1 actively recruited lipid droplets,leading to fusion events and increased cellular lipid consumption.These drug behaviors confirmed MOS-1 is a candidate for the treatment of lipid-related diseases.Furthermore,in a high-fat HepG2 cell model and in high-fat diet-fed apolipoprotein E(ApoE)^(-/-)mice,MOS-1 significantly promoted triglyceride degradation,reduced lipid droplet accumulation,lowered serum triglyceride levels,and mitigated liver damage and steatosis.Overall,our work supports the prioritization of in situ visual monitoring of drug location and distribution in subcellular compartments during the drug development phase,as this methodology contributes to the rapid identification of drug indications.Collectively,this methodology is significant for the screening and development of selective small-molecule drugs,and is expected to expedite the identification of candidate molecules with medicinal effects.
基金Supported by the Jiangsu Coast Development Group Co.,Ltd.the Marine Biological High Value Development and Utilization Project(No.2023YHTZZZ04)+1 种基金the Natural Science Foundation of Shandong Province(No.ZR2022MD007)the Science and Technology Project of Fujian Province(Nos.2022T3024,2023T3057)。
文摘Alginate oligosaccharides(AOS)and fucoidan are two important bioactive carbohydrate which that can be prepared from brown seaweeds.We investigated a novel enzyme-assisted one-pot method on AOS and fucoidan extraction from Lessonia trabeculata,and found that the optimum extraction procedure was:first,using 10-g/L Na_2CO_3 solution to digest the seaweeds for 2 h at 80℃ with the material-liquid ratio 1?20(g/mL),and then adding 4.0%fucoidan cleavage enzyme for reaction for 12 h at 45℃.The yield of AOS and fucoidan was 21.36%and 14.40%,respectively,which was higher than those using traditional methods.The degree of polymerization of AOS was determined to be 3-5,confirming the results through thin layer chromatography(TLC)and electrospray ionization mass spectrometry(ESI-MS).The ratio of guluronic acid to mannuronic acid in AOS was found to be 1.97.Through anion-exchange column chromatography,four sulfated polysaccharide fractions(F0.5,F1.0,F1.5,and F2.0)were successfully isolated from fucoidan.Chemical analysis revealed that fucoidan and F0.5 were heteropolysaccharides,consisting mainly of mannuronic acid,fucose,and xylose,while the other fractions were predominantly composed of fucose and galactose.The sulfate group content increased from 8.74%to 18.76%,while the uronic acid content decreased from 23.79%to 5.58%across the fractions.All fractions of fucoidan exhibited significant antioxidant activity in various tests.There was a positive correlation between the sulfate content and the ability to scavenge superoxide radicals.Additionally,both AOS and the fucoidan fractions showed immunostimulation activity and were nontoxic to RAW264.7 cells.These results indicate that the novel enzyme-assisted one-pot method is a green and efficient approach for the extraction of fucoidan and alginate oligosaccharides from brown seaweeds.It has potential applications in the processing industry of brown seaweeds.
基金This research was funded and supported by the National Natural Science Foundation of China(Grant Number 32001443)Zhengzhou Major Science and Technology Innovation Project of Henan Province of China(Grant Number 2020CXZX0085)Science and Technology Inovation Team of Henan Academy of Agricultural Sciences(Grant Number 2024TD2).
文摘Alginate oligosaccharides(AOS)enhance drought resistance in wheat(Triticum aestivum L.),but the definite mechanisms remain largely unknown.The physiological and transcriptome responses of wheat seedlings treated with AOS were analyzed under drought stress simulated with polyethylene glycol-6000.The results showed that AOS promoted the growth of wheat seedlings and reduced oxidative damage by improving peroxidase and superoxide dismutase activities under drought stress.A total of 10,064 and 15,208 differentially expressed unigenes(DEGs)obtained from the AOS treatment and control samples at 24 and 72 h after dehydration,respectively,were mainly enriched in the biosynthesis of secondary metabolites(phenylpropanoid biosynthesis,flavonoid biosynthesis),carbohydrate metabolism(starch and sucrose metabolism,carbon fixation in photosynthetic organisms),lipid metabolism(fatty acid elongation,biosynthesis of unsaturated fatty acids,alpha-linolenic acid metabolism,cutin,suberine and wax biosynthesis),and signaling transduction pathways.The up-regulated genes were related to,for example,chlorophyll a-b binding protein,amylosynthease,phosphotransferase,peroxidase,phenylalanine ammonia lyase,flavone synthase,glutathione synthetase.Signaling molecules(including MAPK,plant hormones,H_(2)O_(2) and calcium)and transcription factors(mainly including NAC,MYB,MYB-related,WRKY,bZIP family members)were involved in the AOS-induced wheat drought resistance.The results obtained in this study help underpin the mechanisms of wheat drought resistance improved by AOS,and provides a theoretical basis for the application of AOS as an environmentally sustainable biological method to improve drought resistance in agriculture.
文摘利用实际航摄资料评价了WGS 84和国家80坐标系下机载POS(position and orientation system)系统直接对地目标定位的精度,分析了不同大地水准面拟合方法对高程精度的影响,检校了POS系统的视准轴误差,讨论了检校场的检校结果。试验表明,基于POS系统的航空遥感直接对地目标定位至少需要利用带有1个平高地面控制点的检校场对POS系统进行检校才能消除系统误差;在WGS84坐标系中,可以获得较高的定位精度,而转换到国家80坐标系下,必须对高程进行大地水准面拟合改正。
