Glyphosate, the most extensively used herbicide globally, has raised ecotoxicological concerns because it can be transported into the aquatic environment and cause adverse effects on the aquatic system. However, the f...Glyphosate, the most extensively used herbicide globally, has raised ecotoxicological concerns because it can be transported into the aquatic environment and cause adverse effects on the aquatic system. However, the functional mechanism of glyphosate on cyanobacteria are not completely disentangled. In this study, we selected six common cyanobacteria to evaluate glyphosate effects on cyanobacterial growth in monoculture experiment. Results showed that the growth of five tested cyanobacterial species were promoted under different degrees, and only Pseudanabaena was inhibited by glyphosate. In the phylogenetic tree based on gene sequences of 5-enol-pyruvylshikimate-3-phosphate synthase(EPSPS),a target for glyphosate, we found that the position of Pseudanabaena is the closest to plant,which was sensitive to glyphosate, thereby explaining the inhibitory effect of Pseudanabaena following glyphosate exposure. The primary degraded metabolites or analogs did not induce cyanobacterial growth, laterally demonstrating that glyphosate was used as a source of phosphorus to accelerate cyanobacterial growth because phosphorus levels increased in the medium of glyphosate treatment. Overall, this study provides a better understanding of the influence of glyphosate on the composition of aquatic microbiota and explains the mechanism of cyanobacterial response to glyphosate.展开更多
Aging decreases muscle mass,strength,and functional capacity.High-salt stress seems to promote muscle aging and decrease lifespan.However,exercise delays muscle aging and increases longevity,and it may protect muscle ...Aging decreases muscle mass,strength,and functional capacity.High-salt stress seems to promote muscle aging and decrease lifespan.However,exercise delays muscle aging and increases longevity,and it may protect muscle from rapid aging induced by high-salt intake(HSI),but the molecular mechanisms are poorly understood.In this study,the flies were fed a high-salt diet and trained to exercise.Muscle CG2196(salt)gene and dSir2 gene were over-expressed by building mef2-gal4/UAS system.The results showed that both physical exercise and muscle dSir2 gene overexpression prevented HSI-induced and muscle salt overexpression-induced accelerated age-related decline of climbing index,climbing endurance,muscle NAD^(+)level,SOD activity level,dSir2 expression,and dFOXO expression,and they also prevented HSI-induced and muscle salt overexpression-induced accelerated age-related increase in muscle ROS level,MDA level,and salt gene expression.Physical exercise improved lifespan decrease induced by HSI and muscle salt overexpression.Therefore,current results indicated that high-salt stress accelerated muscle aging by decreasing muscular NAD^(+)/dSir2/dFOXO pathway activity and increasing oxidative stress.Physical exercise protected muscle from accelerated aging induced by high-salt stress through activating muscle NAD^(+)/dSir2/dFOXO pathway and enhancing muscle oxidation resistance.The combination of exercise and muscle dSir2 overexpression had the best protective effect on muscle aging and lifespan in flies.展开更多
Background Cognitive impairment represents a major comorbidity among older adults with epilepsy.This study aimed to explore the association between the apolipoprotein E(APOE)ε4 allele and cognitive function in older ...Background Cognitive impairment represents a major comorbidity among older adults with epilepsy.This study aimed to explore the association between the apolipoprotein E(APOE)ε4 allele and cognitive function in older people with epilepsy.Methods People with epilepsy aged≥50 years were enrolled at an outpatient clinic of epilepsy from November 2019 to July 2024.Blood samples were collected for APOE genotyping.Participants were categorized into two groups based on the presence of the APOEε4 allele:APOEε4(+/-).Cognitive function was assessed using a battery with neuropsychologicaltests.Based on Mini-Mental State Examination(MMSE)scores,participants were defned as unimpaired cognition(UC)(MMSE≥27)and cognitive impairment(CI)(MMSE<27).Seizure frequency was categorized into low(≤3/year)and high(>3/year)groups.Multivariate logistic regression analysis and general linear models were employed to identify factors associated with cognitive function.Results Among 110 participants,51(46.4%)were defned as CI.Compared with UC group,the CI group was older(65.1±7.6 vs 60.8±6.8 years,P=0.002),with lower educational level(9.0[7.0,11.0]vs 12.0[9.0,13.0]years,P<0.001),and higher seizure frequency(12.0[1.0,24.0]vs 1.0[0.0,12.0]times/year,P=0.005).High seizure frequency(OR=3.94,95%CI[1.34,11.61],P=0.013)and more APOEε4 alleles(OR=3.28,95%CI[1.09,9.83],P=0.034)were risk factors for CI.An interactive efect between the number of APOEε4 alleles and seizure frequency was observed(P=0.002).Compared to participants with APOEε4(-)and low seizure frequency,those with APOEε4(-)and high seizure frequency showed a threefold risk of CI(OR=3.34,95%CI[0.99,11.25],P=0.051),while those with APOEε4(+)and high frequency demonstrated the highest risk of CI(OR=10.53,95%CI[1.75,63.47],P=0.010).Conclusions The synergistic efect of APOEε4 allele and seizure frequency on cognitive function suggested their importance in clinical assessments and therapeutic approaches in managing older people with epilepsy.展开更多
Inspired by bionic surface design principles,this study developed a one-step method to deposit thiol-isocyanate polymer microspheres on dopamine-treated carbon fiber surfaces,creating a raspberry-like surface architec...Inspired by bionic surface design principles,this study developed a one-step method to deposit thiol-isocyanate polymer microspheres on dopamine-treated carbon fiber surfaces,creating a raspberry-like surface architecture.The bioinspired structure serves as both physical and chemical“bridges”between carbon fibers and epoxy resin,while simultaneously enhancing the chemical activity,mechanical interlocking effect,and wettability of the fiber surfaces.Compared to untreated samples,the modified composites exhibited 63.11%improvement in interlaminar fracture toughness,with impact strength and tensile strength increasing remarkably by 167.55%and 73.59%,respectively.Furthermore,systematic investigation of the interfacial reinforcement mechanisms and energy dissipation modes was conducted through comprehensive fracture morphology analysis.展开更多
Silicosis is the most prevalent and fatal occupational disease with no effective therapeutics,and currently used drugs cannot reverse the disease progress.Worse still,there are still challenges to be addressed to full...Silicosis is the most prevalent and fatal occupational disease with no effective therapeutics,and currently used drugs cannot reverse the disease progress.Worse still,there are still challenges to be addressed to fully decipher the intricated pathogenesis.Thus,specifying the essential mechanisms and targets in silicosis progression then exploring anti-silicosis pharmacuticals are desperately needed.In this work,multi-omics atlas was constructed to depict the pivotal abnormalities of silicosis and develop targeted agents.By utilizing an unbiased and time-resolved analysis of the transcriptome,proteome and phosphoproteome of a silicosis mouse model,we have verified the significant differences in transcript,protein,kinase activity and signaling pathway level during silicosis progression,in which the importance of essential biological processes such as macrophage activation,chemotaxis,immune cell recruitment and chronic inflammation were emphasized.Notably,the phosphorylation of EGFR(p-EGFR)and SYK(pSYK)were identified as potential therapeutic targets in the progression of silicosis.To inhibit and validate these targets,we tested fostamatinib(targeting SYK)and Gefitinib(targeting EGFR),and both drugs effectively ameliorated pulmonary dysfunction and inhibited the progression of inflammation and fibrosis.Overall,our drug discovery with multi-omics approach provides novel and viable therapeutic strategies for the treatment of silicosis.展开更多
Tumor relapse and metastasis are the major causes of mortality associated with urothelial cancer.In the tumor microenvironment,negative regulatory molecules and various immune cell subtypes suppress antitumor immunity...Tumor relapse and metastasis are the major causes of mortality associated with urothelial cancer.In the tumor microenvironment,negative regulatory molecules and various immune cell subtypes suppress antitumor immunity.The inflammatory microenvironment,associated with neutrophils and neutrophil extracellular traps(NETs),promotes tumor metastasis.However,no drugs are currently available to specifically inhibit neutrophils and NETs.In this study,we first demonstrated that icaritin(ICT),a Chinese herbal remedy that is a first-line treatment for advanced and incurable hepatocellular carcinoma,reduces NETs caused by suicidal NETosis and prevents neutrophil infiltration in the tumor microenvironment.Mechanistically,ICT binds to and inhibits the expression of PADI2 in neutrophils,thereby suppressing PADI2-mediated histone citrullination.Moreover,ICT inhibits ROS generation,suppresses the MAPK signaling pathway,and inhibits NET-induced tumor metastasis.Simultaneously,ICT inhibits tumoral PADI2-mediated histone citrullination,which consequently suppresses the transcription of neutrophil-recruiting genes such as GM-CSF and IL-6.The downregulation of IL-6 expression,in turn,forms a regulatory feedback loop through the JAK2/STAT3/IL-6 axis.Through a retrospective study of clinical samples,we found a correlation between neutrophils,NETs,UCa prognosis,and immune evasion.Combining ICT with immune checkpoint inhibitors may have synergistic effects.In summary,our study demonstrated that ICT could be a novel inhibitor of NETs and a novel UCa treatment.展开更多
Locoregional therapy is playing an increasingly important role in the non-surgical management of hepatocellular carcinoma(HCC).The novel technique of non-thermal electric ablation by nanosecond pulsed electric field h...Locoregional therapy is playing an increasingly important role in the non-surgical management of hepatocellular carcinoma(HCC).The novel technique of non-thermal electric ablation by nanosecond pulsed electric field has been recognized as a potential locoregional methodology for indicated HCC.This manuscript explores the most recent studies to indicate its unique anti-tumor immune response.The possible immune mechanism,termed as nano-pulse stimulation,was also analyzed.展开更多
This study prepared buckwheat protein(BP)/soybean soluble polysaccharides(SSPS)nanoparticles for PTS encapsulation,exhibiting nanocomplexes with sizes of under 300 nm,spherical particles and strong negative charge(-20...This study prepared buckwheat protein(BP)/soybean soluble polysaccharides(SSPS)nanoparticles for PTS encapsulation,exhibiting nanocomplexes with sizes of under 300 nm,spherical particles and strong negative charge(-20~-40 mV).Moreover,the addition of SSPS enhanced the embedding effect of PTS from 61.38% to 89.78%.X-ray and fluorescence spectroscopy revealed that PTS can bind to BP and SSPS.Molecular docking analysis showed PTS can bind to BP with hydrogen bonds and hydrophobic bonds at pH 7.0,only can bind with hydrophobic bonds at pH 4.5.Stability experiments revealed that nanocomplex had higher physical and foam stability.Antioxidant experiments revealed that antioxidant capacity of PTS in the nanocomplex was significantly higher than that in the free state.Therefore,the research findings can be advantageous to the utilization of PTS in functional foods.展开更多
Chia seed(CS)expeller was hydrolyzed by 6 proteases(Alcalase,Neutrase,Protamex,Papain,Flavourzyme,Trypsin)in different hydrolytic patterns to improve bioavailability and explore the structure-enzyme active site relati...Chia seed(CS)expeller was hydrolyzed by 6 proteases(Alcalase,Neutrase,Protamex,Papain,Flavourzyme,Trypsin)in different hydrolytic patterns to improve bioavailability and explore the structure-enzyme active site relationship.At the meantime,in silico hydrolysis was performed to obtain a series of proteolytic sequences of chia seed protein treated by different enzymes.The hydrolysates of non-animal derived protease were further determined by in vitro experiment in terms of health-promoting activities.It is necessary to remove fiber before protease hydrolysis,and Alcalase and Flavorzyme had a good effect on the degradation of residual fiber of CS meal.Due to the diversity of enzyme cleave sites,the structure properties of hydrolysate treated by different proteases exhibited discrepancy.Furthermore,hydrolysates of CS meal presented varying bioactivity(antioxidant,hypoglycemic,hypotensive,hypolipidemic,anti-alcoholic activity).Among them,Neutrase,Protamex and Papain hydrolysis showed significant impact on the production of multiple active functional factors.The highest percentual of angiotensin I converting enzyme inhibitory(ACE)and alcohol dehydrogenase activation(ADH)effect were observed in Neutrase and Flavourzyme hydrolysates,with lowest IC50 of 0.219±0.030 mg/mL and 0.047±0.002 mg/mL,respectively.Ultimately,an activity-oriented protease screening method was obtained,which consists of in silico digestion and in vitro activity evaluation.展开更多
In this study,the effects of different enzyme deactivation methods(80,100,4℃ and pH 6.0)on the structural characteristics,bound polyphenol content and antioxidant activity of buckwheat protein hydrolyzate-phenol comp...In this study,the effects of different enzyme deactivation methods(80,100,4℃ and pH 6.0)on the structural characteristics,bound polyphenol content and antioxidant activity of buckwheat protein hydrolyzate-phenol complex(BHP)were investigated.Alcalase was selected for enzymolysis.Amino acid analysis showed that glutamic acid was the most important amino acid in hydrolysate.Molecular weight distribution analysis showed that the hydrolysis can be completed at 3 h.After enzyme deactivation treatment,all hydrolysates were dialyzed to remove aggregates/enzymes(15 kDa).Fourier transform infrared spectroscopy(FTIR)analysis revealed that the characteristic peaks of BHP shifted after treatment.The fluorescence spectra and surface hydrophobicity(Ho)showed that the corresponding values of BHP increased significantly when deactivated at 80℃ and pH 6.0,decreased at 100℃ and remained no change at 4℃.The content of total bound polyphenol increased when treated at 80 and 4℃,and decreased at 100℃ and pH 6.0.Meanwhile,the results showed that all BHP have high antioxidant capacity.This work can provide more support for the systematic evaluation of enzyme deactivation methods containing phenolic hydrolysates.展开更多
基金supported by the National Natural Science Foundation of China (Nos. 21976161, 21777144, 41907210)。
文摘Glyphosate, the most extensively used herbicide globally, has raised ecotoxicological concerns because it can be transported into the aquatic environment and cause adverse effects on the aquatic system. However, the functional mechanism of glyphosate on cyanobacteria are not completely disentangled. In this study, we selected six common cyanobacteria to evaluate glyphosate effects on cyanobacterial growth in monoculture experiment. Results showed that the growth of five tested cyanobacterial species were promoted under different degrees, and only Pseudanabaena was inhibited by glyphosate. In the phylogenetic tree based on gene sequences of 5-enol-pyruvylshikimate-3-phosphate synthase(EPSPS),a target for glyphosate, we found that the position of Pseudanabaena is the closest to plant,which was sensitive to glyphosate, thereby explaining the inhibitory effect of Pseudanabaena following glyphosate exposure. The primary degraded metabolites or analogs did not induce cyanobacterial growth, laterally demonstrating that glyphosate was used as a source of phosphorus to accelerate cyanobacterial growth because phosphorus levels increased in the medium of glyphosate treatment. Overall, this study provides a better understanding of the influence of glyphosate on the composition of aquatic microbiota and explains the mechanism of cyanobacterial response to glyphosate.
基金This work is supported by the National Natural Science Foundation of China(NSFC)(32000832)Province Natural Science Foundation of Shandong(ZR2020QC096).
文摘Aging decreases muscle mass,strength,and functional capacity.High-salt stress seems to promote muscle aging and decrease lifespan.However,exercise delays muscle aging and increases longevity,and it may protect muscle from rapid aging induced by high-salt intake(HSI),but the molecular mechanisms are poorly understood.In this study,the flies were fed a high-salt diet and trained to exercise.Muscle CG2196(salt)gene and dSir2 gene were over-expressed by building mef2-gal4/UAS system.The results showed that both physical exercise and muscle dSir2 gene overexpression prevented HSI-induced and muscle salt overexpression-induced accelerated age-related decline of climbing index,climbing endurance,muscle NAD^(+)level,SOD activity level,dSir2 expression,and dFOXO expression,and they also prevented HSI-induced and muscle salt overexpression-induced accelerated age-related increase in muscle ROS level,MDA level,and salt gene expression.Physical exercise improved lifespan decrease induced by HSI and muscle salt overexpression.Therefore,current results indicated that high-salt stress accelerated muscle aging by decreasing muscular NAD^(+)/dSir2/dFOXO pathway activity and increasing oxidative stress.Physical exercise protected muscle from accelerated aging induced by high-salt stress through activating muscle NAD^(+)/dSir2/dFOXO pathway and enhancing muscle oxidation resistance.The combination of exercise and muscle dSir2 overexpression had the best protective effect on muscle aging and lifespan in flies.
基金sponsored by the coordinating project of neurology department of Huashan Hospital[3030512199]the National Natural Science Foundation of China[82071456]+1 种基金Shanghai Academic Research Leader Program[21XD1420900]Shanghai Municipal Commission of Health Program[20224Z0002].
文摘Background Cognitive impairment represents a major comorbidity among older adults with epilepsy.This study aimed to explore the association between the apolipoprotein E(APOE)ε4 allele and cognitive function in older people with epilepsy.Methods People with epilepsy aged≥50 years were enrolled at an outpatient clinic of epilepsy from November 2019 to July 2024.Blood samples were collected for APOE genotyping.Participants were categorized into two groups based on the presence of the APOEε4 allele:APOEε4(+/-).Cognitive function was assessed using a battery with neuropsychologicaltests.Based on Mini-Mental State Examination(MMSE)scores,participants were defned as unimpaired cognition(UC)(MMSE≥27)and cognitive impairment(CI)(MMSE<27).Seizure frequency was categorized into low(≤3/year)and high(>3/year)groups.Multivariate logistic regression analysis and general linear models were employed to identify factors associated with cognitive function.Results Among 110 participants,51(46.4%)were defned as CI.Compared with UC group,the CI group was older(65.1±7.6 vs 60.8±6.8 years,P=0.002),with lower educational level(9.0[7.0,11.0]vs 12.0[9.0,13.0]years,P<0.001),and higher seizure frequency(12.0[1.0,24.0]vs 1.0[0.0,12.0]times/year,P=0.005).High seizure frequency(OR=3.94,95%CI[1.34,11.61],P=0.013)and more APOEε4 alleles(OR=3.28,95%CI[1.09,9.83],P=0.034)were risk factors for CI.An interactive efect between the number of APOEε4 alleles and seizure frequency was observed(P=0.002).Compared to participants with APOEε4(-)and low seizure frequency,those with APOEε4(-)and high seizure frequency showed a threefold risk of CI(OR=3.34,95%CI[0.99,11.25],P=0.051),while those with APOEε4(+)and high frequency demonstrated the highest risk of CI(OR=10.53,95%CI[1.75,63.47],P=0.010).Conclusions The synergistic efect of APOEε4 allele and seizure frequency on cognitive function suggested their importance in clinical assessments and therapeutic approaches in managing older people with epilepsy.
基金supported by the National Natural Science Foundation of China(No.52505378)the Jilin Provincial Scientific and Technological Department(20250301045YY).
文摘Inspired by bionic surface design principles,this study developed a one-step method to deposit thiol-isocyanate polymer microspheres on dopamine-treated carbon fiber surfaces,creating a raspberry-like surface architecture.The bioinspired structure serves as both physical and chemical“bridges”between carbon fibers and epoxy resin,while simultaneously enhancing the chemical activity,mechanical interlocking effect,and wettability of the fiber surfaces.Compared to untreated samples,the modified composites exhibited 63.11%improvement in interlaminar fracture toughness,with impact strength and tensile strength increasing remarkably by 167.55%and 73.59%,respectively.Furthermore,systematic investigation of the interfacial reinforcement mechanisms and energy dissipation modes was conducted through comprehensive fracture morphology analysis.
基金funded by Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences(CIFMS)(grant number:2021-I2M-1-049)National Key Research and Development Program of China Grants(grant numbers:2021YFC2500700)the Non-profit Central Research Institute Fund of Chinese Academy of Medical Sciences(grant number:2021RC31002,2018RC31001).
文摘Silicosis is the most prevalent and fatal occupational disease with no effective therapeutics,and currently used drugs cannot reverse the disease progress.Worse still,there are still challenges to be addressed to fully decipher the intricated pathogenesis.Thus,specifying the essential mechanisms and targets in silicosis progression then exploring anti-silicosis pharmacuticals are desperately needed.In this work,multi-omics atlas was constructed to depict the pivotal abnormalities of silicosis and develop targeted agents.By utilizing an unbiased and time-resolved analysis of the transcriptome,proteome and phosphoproteome of a silicosis mouse model,we have verified the significant differences in transcript,protein,kinase activity and signaling pathway level during silicosis progression,in which the importance of essential biological processes such as macrophage activation,chemotaxis,immune cell recruitment and chronic inflammation were emphasized.Notably,the phosphorylation of EGFR(p-EGFR)and SYK(pSYK)were identified as potential therapeutic targets in the progression of silicosis.To inhibit and validate these targets,we tested fostamatinib(targeting SYK)and Gefitinib(targeting EGFR),and both drugs effectively ameliorated pulmonary dysfunction and inhibited the progression of inflammation and fibrosis.Overall,our drug discovery with multi-omics approach provides novel and viable therapeutic strategies for the treatment of silicosis.
基金grants from National Natural Science Foundation of China(No.82373222,No.82202970)Leading Talent Program by Shanghai Municipal Health Commission(2022LJ008,China)+1 种基金Shanghai Shenkang Hospital Development Center(SHDC12021104,China)Science and Technology Commission of Shanghai Municipality(22Y21900200&22YF1404300,China).
文摘Tumor relapse and metastasis are the major causes of mortality associated with urothelial cancer.In the tumor microenvironment,negative regulatory molecules and various immune cell subtypes suppress antitumor immunity.The inflammatory microenvironment,associated with neutrophils and neutrophil extracellular traps(NETs),promotes tumor metastasis.However,no drugs are currently available to specifically inhibit neutrophils and NETs.In this study,we first demonstrated that icaritin(ICT),a Chinese herbal remedy that is a first-line treatment for advanced and incurable hepatocellular carcinoma,reduces NETs caused by suicidal NETosis and prevents neutrophil infiltration in the tumor microenvironment.Mechanistically,ICT binds to and inhibits the expression of PADI2 in neutrophils,thereby suppressing PADI2-mediated histone citrullination.Moreover,ICT inhibits ROS generation,suppresses the MAPK signaling pathway,and inhibits NET-induced tumor metastasis.Simultaneously,ICT inhibits tumoral PADI2-mediated histone citrullination,which consequently suppresses the transcription of neutrophil-recruiting genes such as GM-CSF and IL-6.The downregulation of IL-6 expression,in turn,forms a regulatory feedback loop through the JAK2/STAT3/IL-6 axis.Through a retrospective study of clinical samples,we found a correlation between neutrophils,NETs,UCa prognosis,and immune evasion.Combining ICT with immune checkpoint inhibitors may have synergistic effects.In summary,our study demonstrated that ICT could be a novel inhibitor of NETs and a novel UCa treatment.
基金This study was funded by the Xinjiang Key Lab Project(No.2014KL002 to X Chen)the National Natural Science Foundation of China(No.81372425 to X Chen)+1 种基金National S&T Major Project(No.SQ2018ZX100301 to X Chen)the Zhejiang Natural Science Foundation(No.LY17H160018 to X Miao)
文摘Locoregional therapy is playing an increasingly important role in the non-surgical management of hepatocellular carcinoma(HCC).The novel technique of non-thermal electric ablation by nanosecond pulsed electric field has been recognized as a potential locoregional methodology for indicated HCC.This manuscript explores the most recent studies to indicate its unique anti-tumor immune response.The possible immune mechanism,termed as nano-pulse stimulation,was also analyzed.
基金financially supported by the National Key Research and Development Project of China(No.2016YFD0401204)National Natural Science Foundation of China(No.31671891).
文摘This study prepared buckwheat protein(BP)/soybean soluble polysaccharides(SSPS)nanoparticles for PTS encapsulation,exhibiting nanocomplexes with sizes of under 300 nm,spherical particles and strong negative charge(-20~-40 mV).Moreover,the addition of SSPS enhanced the embedding effect of PTS from 61.38% to 89.78%.X-ray and fluorescence spectroscopy revealed that PTS can bind to BP and SSPS.Molecular docking analysis showed PTS can bind to BP with hydrogen bonds and hydrophobic bonds at pH 7.0,only can bind with hydrophobic bonds at pH 4.5.Stability experiments revealed that nanocomplex had higher physical and foam stability.Antioxidant experiments revealed that antioxidant capacity of PTS in the nanocomplex was significantly higher than that in the free state.Therefore,the research findings can be advantageous to the utilization of PTS in functional foods.
基金the supports of Major Science and Technology Special Project of Yunnan Province“Major Special Projects of Bio-Breeding Industry and Deep Processing of Agricultural Products”(NO:202102AE090055)Major Science and Technology Special Project of Yunnan Province“Major Special Projects of Bio-Breeding Industry and Deep Processing of Agricultural Products”(NO:2019JZZY010708).
文摘Chia seed(CS)expeller was hydrolyzed by 6 proteases(Alcalase,Neutrase,Protamex,Papain,Flavourzyme,Trypsin)in different hydrolytic patterns to improve bioavailability and explore the structure-enzyme active site relationship.At the meantime,in silico hydrolysis was performed to obtain a series of proteolytic sequences of chia seed protein treated by different enzymes.The hydrolysates of non-animal derived protease were further determined by in vitro experiment in terms of health-promoting activities.It is necessary to remove fiber before protease hydrolysis,and Alcalase and Flavorzyme had a good effect on the degradation of residual fiber of CS meal.Due to the diversity of enzyme cleave sites,the structure properties of hydrolysate treated by different proteases exhibited discrepancy.Furthermore,hydrolysates of CS meal presented varying bioactivity(antioxidant,hypoglycemic,hypotensive,hypolipidemic,anti-alcoholic activity).Among them,Neutrase,Protamex and Papain hydrolysis showed significant impact on the production of multiple active functional factors.The highest percentual of angiotensin I converting enzyme inhibitory(ACE)and alcohol dehydrogenase activation(ADH)effect were observed in Neutrase and Flavourzyme hydrolysates,with lowest IC50 of 0.219±0.030 mg/mL and 0.047±0.002 mg/mL,respectively.Ultimately,an activity-oriented protease screening method was obtained,which consists of in silico digestion and in vitro activity evaluation.
基金supported by Major Science and Technology Special Project of Yunnan Province“Major Special Projects of Bio-Breeding Industry and Deep Processing of Agricultural Products”(NO:202102AE090055).
文摘In this study,the effects of different enzyme deactivation methods(80,100,4℃ and pH 6.0)on the structural characteristics,bound polyphenol content and antioxidant activity of buckwheat protein hydrolyzate-phenol complex(BHP)were investigated.Alcalase was selected for enzymolysis.Amino acid analysis showed that glutamic acid was the most important amino acid in hydrolysate.Molecular weight distribution analysis showed that the hydrolysis can be completed at 3 h.After enzyme deactivation treatment,all hydrolysates were dialyzed to remove aggregates/enzymes(15 kDa).Fourier transform infrared spectroscopy(FTIR)analysis revealed that the characteristic peaks of BHP shifted after treatment.The fluorescence spectra and surface hydrophobicity(Ho)showed that the corresponding values of BHP increased significantly when deactivated at 80℃ and pH 6.0,decreased at 100℃ and remained no change at 4℃.The content of total bound polyphenol increased when treated at 80 and 4℃,and decreased at 100℃ and pH 6.0.Meanwhile,the results showed that all BHP have high antioxidant capacity.This work can provide more support for the systematic evaluation of enzyme deactivation methods containing phenolic hydrolysates.
