Trehalose is an autophagy-promoting disaccharide,which can improve and delay chronic diseases like neurodegenerative diseases and atherosclerosis,but its bioavailability is severely restricted by endogenous trehalase ...Trehalose is an autophagy-promoting disaccharide,which can improve and delay chronic diseases like neurodegenerative diseases and atherosclerosis,but its bioavailability is severely restricted by endogenous trehalase in mammals.Trehalase inhibitor is a promising and effective way to enhance trehalose bioavailability by preventing trehalose from hydrolyzing.However,previously reported trehalase inhibitors still face safety of long-term use and promiscuous inhibition on intestinal glycosidases.This study carried out a high-throughput virtual screening through molecular pool-based molecular docking combined with in vitro inhibition experiments to screen trehalase inhibitors naturally derived from foods.Out of 1769 small molecules,which include 115 analogs of trehalose,natural monosaccharides,disaccharides,trisaccharides,imidazoles and their derivatives,as well as 20 natural amino acids and their 400 dipeptides,isomaltose,α-isomaltulose,and isomaltitol exhibited the best inhibitory activities,beyond as traditional sweetener and prebiotic.Best of all,isomaltose showed the half maximal inhibitory concentration(IC50)and inhibition constant(Ki)values on trehalase of 5.59 and(2.1760±0.3431)mmol/L,respectively.Moreover,isomaltose was resistant to the simulated digestive environment and did not affect intestinal glycosidases such asα-glucosidase and glucoamylase,making it a reliable edible candidate for a trehalase inhibitor.This study provides new insights into the virtual screening-based identification of new food-derived trehalase inhibitors for enhanced integrity and bioavailability of orally administered trehalose,especially repurposing a prebiotic for another new use as trehalase inhibitor.展开更多
Trehalase hydrolyzes trehalose to glucose to provide energy for insects or building blocks for chitin synthesis.Because trehalase is critical to insects but not to humans,it has long been considered a promising target...Trehalase hydrolyzes trehalose to glucose to provide energy for insects or building blocks for chitin synthesis.Because trehalase is critical to insects but not to humans,it has long been considered a promising target for green insecticides.However,the known trehalase inhibitors are mainly sugar derivatives with poor druggability.In this study,the trehalase from Ostrinia furnacalis(OfTreh)was expressed and characterized.By integrative computational strategies,diphenyl ether herbicides were discovered as the first non-carbohydrate inhibitors of insect trehalases.Bifenox and its more stable derivative,chlomethoxyfen,inhibited Of Treh with Ki values of 56 and 43μM,respectively.The oral administration of bifenox or chlomethoxyfen to locusts resulted in the inhibition of trehalose hydrolysis in vivo,leading to a mortality rate of 66%and server locomotion disorder in the survivors.This study not only established a platform for the development of insecticides targeting trehalase but also discovered a new mechanism for diphenyl ethers to kill insects as trehalase inhibitors.展开更多
The Asian citrus psyllid,Diaphorina citri is the principal vector of huanglongbing,which transmits Candidatus Liberibacter asiaticus.Trehalase is a key enzyme involved in trehalose hydrolysis and plays an important ro...The Asian citrus psyllid,Diaphorina citri is the principal vector of huanglongbing,which transmits Candidatus Liberibacter asiaticus.Trehalase is a key enzyme involved in trehalose hydrolysis and plays an important role in insect growth and development.The specific functions of this enzyme in D.citri have not been determined.In this study,three trehalase genes(DcTreJ-1,DcTrel-2,and DcTre2)were identified based on the D.citri genome database.Bioinformatic analysis showed that DcTrel-1 and DcTrel-2 are related to soluble trehalase,whereas DcTre2 is associated with membrane-bound trehalase.Spatiotemporal expression analysis indicated that DcTrel-1 and DcTrel-2 had the highest expression levels in the head and wing,respectively,and DcTre2 had high expression levels in the fat body.Furthermore,DcTrel-1 and DcTrel-2 expression levels were induced by 20-hydroxyecdysone and juvenile hormone III,but DcTre2 was unaffected.The expression levels of DcTrel-1,DcTrel-2,and DcTre2 were significantly up-regulated,which resulted in high mortality after treatment with validamycin.Trehalase activities and glucose contents were downregulated,but the trehalose content increased after treatment with validamycin.In addition,the expression levels of chitin metabolism-related genes significantly decreased at 24 and 48 h after treatment with validamycin.Furthermore,silencing of DcTrel-1,DcTrel-2,and DcTre2 reduced the expression levels of chitin metabolism-related genes and led to a malformed phenotype of D.citri.These results indicate that D.citri trehalase plays an essential role in regulating chitin metabolism and provides a new target for control of D.citri.展开更多
Apolygus lucorum is the dominant pathogenic insect attacking Bacillus thuringiensis(Bt)cotton in China.Additionally,20-hydroxyecdysone(20E)has important functions in many biological processes,including insect reproduc...Apolygus lucorum is the dominant pathogenic insect attacking Bacillus thuringiensis(Bt)cotton in China.Additionally,20-hydroxyecdysone(20E)has important functions in many biological processes,including insect reproduction.Phospholipase C(PLC),which is an essential enzyme for phosphoinositide metabolism,is involved in 20E signal transduction,but its function in 20E-mediated reproduction in A.lucorum remains unclear.In this study,20E increased A/PLCγ transcription as well as the abundance and activity of the encoded protein during molting and metamorphosis.The 20E treatment also induced the considerable accumulation of two second messengers,inositol triphosphate and diacylglycerol.The expression levels of genes encoding vitellogenin(AlVg)and soluble trehalase(AlTre-1)were similar to those of AlPLCy,and were upregulated in response to 20C.The silencing of AlPLCγ resulted in downregulated expression o f AITre-γ smdAlVg.However,the silencing of AlTre-1 and AlVg did not affect AlPLCγ expression.Moreover,the silencing of AlVg did not alter AlTre-1 expression.Furthermore,an examination of the insect specimens indicated that AlPLCy is required for female adult reproduction,and that downregulated expression of this gene is associated with decreases in fecundity,adult longevity,and egg hatching rate as well as delayed oocyte maturation.We propose that 20E regulates AlTre-1 expression via AlPLCy and affects Vg expression as well as ovary development to facilitate the reproductive activities of A.lucorum females.展开更多
Trehalose is a non-reducing disaccharide connected byα-1,1-glycosidic bonds;it is widely distributed in bacteria,fungi,yeast,insects,and plant tissues and plays various roles.It can be hydrolyzed by trehalase into tw...Trehalose is a non-reducing disaccharide connected byα-1,1-glycosidic bonds;it is widely distributed in bacteria,fungi,yeast,insects,and plant tissues and plays various roles.It can be hydrolyzed by trehalase into two glucose molecules.Trehalases from different sources have been expressed in Escherichia coli,Pichia pastoris,Saccharomyces cerevisiae,baculovirus-silkworm,and other expression systems;however,it is most common in E.coli.The structural characteristics of different glycoside hydrolase(GH)family trehalases and the sources of trehalase have been analyzed.The catalytic mechanism of GH37 trehalase has also been elucidated in detail.Moreover,the molecular modification of trehalase has mainly focused on directed evolution for improving enzyme activity.We comprehensively reviewed the current application status and adaptable transformations was comprehensively overviewed in the context of industrial performance.We suggest that the level of recombinant production is far from meeting industrial requirements,and the catalytic performance of trehalase needs to be improved urgently.Finally,we discuss developmental prospects and future trends.展开更多
Background: Immune protection in newborn calves relies on a combination of the timing,volume and quality of colostrum consumed by the calf after birth.Poor quality colostrum with inadequate immunoglobulin concentratio...Background: Immune protection in newborn calves relies on a combination of the timing,volume and quality of colostrum consumed by the calf after birth.Poor quality colostrum with inadequate immunoglobulin concentration contributes to failed transfer of passive immunity in calves,leading to higher calf morbidity and mortality.Therefore,estimating colostrum quality and ensuring the transfer of passive immunity on farm is of critical importance.Currently,there are no on-farm tools that directly measure immunoglobulin content in colostrum or serum.The aim of this study was to apply a novel molecular assay,split trehalase immunoglobulin G assay(STIGA),to directly estimate immunoglobulin content in dairy and beef colostrum and calf sera,and to examine its potential to be developed as on-farm test.The STIGA is based on a split version of trehalase TreA,an enzyme that converts trehalose into glucose,enabling the use of a common glucometer for signal detection.In a first study,60 dairy and64 beef colostrum and 83 dairy and 84 beef calf sera samples were tested with STIGA,and the resulting glucose production was measured and compared with radial immunodiffusion,the standard method for measuring immunoglobulin concentrations.Results: Pearson correlation coefficients between the methods were determined and the sensitivity,specificity,and accuracy of the test were calculated for different colostrum quality and failed transfer of passive immunity cut-off points.The correlations of the STIGA measured by colorimetric enzymatic reaction compared to radial immunodiffusion for dairy and beef colostrum were 0.72 and 0.73,respectively,whereas the correlations for dairy and beef sera were 0.9 and 0.85,respectively.Next,STIGA was tested in a blinded study with fresh colostrum and serum samples where the correlation coefficient was 0.93 and 0.94,respectively.Furthermore,the performance of STIGA followed by glucometer readings resulted in correlations with radial immunodiffusion of 0.7 and 0.85 for dairy and beef colostrum and 0.94 and 0.83 for dairy and beef calf serum.Conclusions: A split TreA assay was validated for measurement of the immunoglobulin content of colostrum and calf sera using both a lab-based format and in a more user-friendly format compatible with on-farm testing.展开更多
基金supported by the National Natural Science Foundation of China(32360564)the Guangxi Science and Technology Major Project(2021AA17011)the Nanning Government Specially-invited Expert Program,and Guangxi University Natural Science and Technological Innovation Development Multiplication Plan Project(2022BZRC010).
文摘Trehalose is an autophagy-promoting disaccharide,which can improve and delay chronic diseases like neurodegenerative diseases and atherosclerosis,but its bioavailability is severely restricted by endogenous trehalase in mammals.Trehalase inhibitor is a promising and effective way to enhance trehalose bioavailability by preventing trehalose from hydrolyzing.However,previously reported trehalase inhibitors still face safety of long-term use and promiscuous inhibition on intestinal glycosidases.This study carried out a high-throughput virtual screening through molecular pool-based molecular docking combined with in vitro inhibition experiments to screen trehalase inhibitors naturally derived from foods.Out of 1769 small molecules,which include 115 analogs of trehalose,natural monosaccharides,disaccharides,trisaccharides,imidazoles and their derivatives,as well as 20 natural amino acids and their 400 dipeptides,isomaltose,α-isomaltulose,and isomaltitol exhibited the best inhibitory activities,beyond as traditional sweetener and prebiotic.Best of all,isomaltose showed the half maximal inhibitory concentration(IC50)and inhibition constant(Ki)values on trehalase of 5.59 and(2.1760±0.3431)mmol/L,respectively.Moreover,isomaltose was resistant to the simulated digestive environment and did not affect intestinal glycosidases such asα-glucosidase and glucoamylase,making it a reliable edible candidate for a trehalase inhibitor.This study provides new insights into the virtual screening-based identification of new food-derived trehalase inhibitors for enhanced integrity and bioavailability of orally administered trehalose,especially repurposing a prebiotic for another new use as trehalase inhibitor.
基金the staff of the BL18U/BL19U1 Beamline of the National Facility for Protein Science,Shanghai,at the Shanghai Synchrotron Radiation Facility for assistance during data collectionthe National Key Research and Development Program of China(Grant No.2023YFD1700500,2022YFD1700200)the Project of Natural Science Foundation of Liaoning Province(2022-KF-15-02).
文摘Trehalase hydrolyzes trehalose to glucose to provide energy for insects or building blocks for chitin synthesis.Because trehalase is critical to insects but not to humans,it has long been considered a promising target for green insecticides.However,the known trehalase inhibitors are mainly sugar derivatives with poor druggability.In this study,the trehalase from Ostrinia furnacalis(OfTreh)was expressed and characterized.By integrative computational strategies,diphenyl ether herbicides were discovered as the first non-carbohydrate inhibitors of insect trehalases.Bifenox and its more stable derivative,chlomethoxyfen,inhibited Of Treh with Ki values of 56 and 43μM,respectively.The oral administration of bifenox or chlomethoxyfen to locusts resulted in the inhibition of trehalose hydrolysis in vivo,leading to a mortality rate of 66%and server locomotion disorder in the survivors.This study not only established a platform for the development of insecticides targeting trehalase but also discovered a new mechanism for diphenyl ethers to kill insects as trehalase inhibitors.
基金the National Natural Science Foundation of China(31960116)the Educational Commission of Jiangxi Province of China(GJJ180747)+1 种基金the National Key Research and Development Program of China(2018YFD0201504,the earmarked fund for Jiangxi Agriculture Research System(JXARS-07)Key Research and Development Plan of Jiangxi Province(20192BBF60057).
文摘The Asian citrus psyllid,Diaphorina citri is the principal vector of huanglongbing,which transmits Candidatus Liberibacter asiaticus.Trehalase is a key enzyme involved in trehalose hydrolysis and plays an important role in insect growth and development.The specific functions of this enzyme in D.citri have not been determined.In this study,three trehalase genes(DcTreJ-1,DcTrel-2,and DcTre2)were identified based on the D.citri genome database.Bioinformatic analysis showed that DcTrel-1 and DcTrel-2 are related to soluble trehalase,whereas DcTre2 is associated with membrane-bound trehalase.Spatiotemporal expression analysis indicated that DcTrel-1 and DcTrel-2 had the highest expression levels in the head and wing,respectively,and DcTre2 had high expression levels in the fat body.Furthermore,DcTrel-1 and DcTrel-2 expression levels were induced by 20-hydroxyecdysone and juvenile hormone III,but DcTre2 was unaffected.The expression levels of DcTrel-1,DcTrel-2,and DcTre2 were significantly up-regulated,which resulted in high mortality after treatment with validamycin.Trehalase activities and glucose contents were downregulated,but the trehalose content increased after treatment with validamycin.In addition,the expression levels of chitin metabolism-related genes significantly decreased at 24 and 48 h after treatment with validamycin.Furthermore,silencing of DcTrel-1,DcTrel-2,and DcTre2 reduced the expression levels of chitin metabolism-related genes and led to a malformed phenotype of D.citri.These results indicate that D.citri trehalase plays an essential role in regulating chitin metabolism and provides a new target for control of D.citri.
基金This study was supported by Chinese Agricultural Research System(CARS-15-18),the Integration Research and Demonstration of the Technology of Cotton Fertilizer and Pesticide Reduction of China(2017YFD0201900)Jiangsu Agricultural Science and Technology Innovation Fund(CX(19)3098)+1 种基金grants from the National Natural Science Fund of China(31301668)JAAS Research Foundation(6111613).
文摘Apolygus lucorum is the dominant pathogenic insect attacking Bacillus thuringiensis(Bt)cotton in China.Additionally,20-hydroxyecdysone(20E)has important functions in many biological processes,including insect reproduction.Phospholipase C(PLC),which is an essential enzyme for phosphoinositide metabolism,is involved in 20E signal transduction,but its function in 20E-mediated reproduction in A.lucorum remains unclear.In this study,20E increased A/PLCγ transcription as well as the abundance and activity of the encoded protein during molting and metamorphosis.The 20E treatment also induced the considerable accumulation of two second messengers,inositol triphosphate and diacylglycerol.The expression levels of genes encoding vitellogenin(AlVg)and soluble trehalase(AlTre-1)were similar to those of AlPLCy,and were upregulated in response to 20C.The silencing of AlPLCγ resulted in downregulated expression o f AITre-γ smdAlVg.However,the silencing of AlTre-1 and AlVg did not affect AlPLCγ expression.Moreover,the silencing of AlVg did not alter AlTre-1 expression.Furthermore,an examination of the insect specimens indicated that AlPLCy is required for female adult reproduction,and that downregulated expression of this gene is associated with decreases in fecundity,adult longevity,and egg hatching rate as well as delayed oocyte maturation.We propose that 20E regulates AlTre-1 expression via AlPLCy and affects Vg expression as well as ovary development to facilitate the reproductive activities of A.lucorum females.
基金supported by the National Key Research and Development Program of China(No.2021YFC2102000)the National Natural Science Foundation of China(No.21978116 and 32171261)the Ningxia Hui Autonomous Region Key Research&Development Plan(No.2019BCH01002).
文摘Trehalose is a non-reducing disaccharide connected byα-1,1-glycosidic bonds;it is widely distributed in bacteria,fungi,yeast,insects,and plant tissues and plays various roles.It can be hydrolyzed by trehalase into two glucose molecules.Trehalases from different sources have been expressed in Escherichia coli,Pichia pastoris,Saccharomyces cerevisiae,baculovirus-silkworm,and other expression systems;however,it is most common in E.coli.The structural characteristics of different glycoside hydrolase(GH)family trehalases and the sources of trehalase have been analyzed.The catalytic mechanism of GH37 trehalase has also been elucidated in detail.Moreover,the molecular modification of trehalase has mainly focused on directed evolution for improving enzyme activity.We comprehensively reviewed the current application status and adaptable transformations was comprehensively overviewed in the context of industrial performance.We suggest that the level of recombinant production is far from meeting industrial requirements,and the catalytic performance of trehalase needs to be improved urgently.Finally,we discuss developmental prospects and future trends.
基金Natural Sciences and Engineering Research Council of Canada,Discovery Grant to JDB
文摘Background: Immune protection in newborn calves relies on a combination of the timing,volume and quality of colostrum consumed by the calf after birth.Poor quality colostrum with inadequate immunoglobulin concentration contributes to failed transfer of passive immunity in calves,leading to higher calf morbidity and mortality.Therefore,estimating colostrum quality and ensuring the transfer of passive immunity on farm is of critical importance.Currently,there are no on-farm tools that directly measure immunoglobulin content in colostrum or serum.The aim of this study was to apply a novel molecular assay,split trehalase immunoglobulin G assay(STIGA),to directly estimate immunoglobulin content in dairy and beef colostrum and calf sera,and to examine its potential to be developed as on-farm test.The STIGA is based on a split version of trehalase TreA,an enzyme that converts trehalose into glucose,enabling the use of a common glucometer for signal detection.In a first study,60 dairy and64 beef colostrum and 83 dairy and 84 beef calf sera samples were tested with STIGA,and the resulting glucose production was measured and compared with radial immunodiffusion,the standard method for measuring immunoglobulin concentrations.Results: Pearson correlation coefficients between the methods were determined and the sensitivity,specificity,and accuracy of the test were calculated for different colostrum quality and failed transfer of passive immunity cut-off points.The correlations of the STIGA measured by colorimetric enzymatic reaction compared to radial immunodiffusion for dairy and beef colostrum were 0.72 and 0.73,respectively,whereas the correlations for dairy and beef sera were 0.9 and 0.85,respectively.Next,STIGA was tested in a blinded study with fresh colostrum and serum samples where the correlation coefficient was 0.93 and 0.94,respectively.Furthermore,the performance of STIGA followed by glucometer readings resulted in correlations with radial immunodiffusion of 0.7 and 0.85 for dairy and beef colostrum and 0.94 and 0.83 for dairy and beef calf serum.Conclusions: A split TreA assay was validated for measurement of the immunoglobulin content of colostrum and calf sera using both a lab-based format and in a more user-friendly format compatible with on-farm testing.
