In 2021,approximately 537 million people suffered from diabetes mellitus(DM)globally,and this figure will increase to approximately 783 million within the next quarter-century.The increasing burden of DM is a pressing...In 2021,approximately 537 million people suffered from diabetes mellitus(DM)globally,and this figure will increase to approximately 783 million within the next quarter-century.The increasing burden of DM is a pressing global public health issue.Therefore,the early identification of high-risk groups and implementation of effective intervention measures is imperative.展开更多
Food safety is a major issue to public health and have attracted global attention.Fast,sensitive,and reliable detection methods for food hazardous substances is highly desirable.Aptamers which can bind to the target m...Food safety is a major issue to public health and have attracted global attention.Fast,sensitive,and reliable detection methods for food hazardous substances is highly desirable.Aptamers which can bind to the target molecules with high affinity and specificity represent an attractive tool for the recognition of food hazardous substances,which play an important role in the development and application of new food safety detection technology.But current assays for characterizing small molecule-aptamer binding are limited by either the mass sensitivity or the size differentiation ability.Herein,we proposed a comprehensive method for assessing the dissociation equilibria of small molecule-aptamer,which is immobilized-free under ambient conditions.The design employs the Le Chatelier’s principle and could be used to effectively measure small molecule-aptamer interactions.ATP binding aptamer and anti-aflatoxin B1 aptamer were used as the model system to determine their affinity,in which their dissociation equilibria measurements are in excellent close to their previous work.Due to the simplicity and sensitivity of this new method,we believe that it could be recommended as an effective tool for characterizing small molecule-aptamer interactions and promote the further application of small molecular aptamer in food safety.展开更多
Circular aptamers are promising candidates for analytical and therapeutic applications due to their enhanced biological and structural stability.However,the process of circular aptamer selection remains a great challe...Circular aptamers are promising candidates for analytical and therapeutic applications due to their enhanced biological and structural stability.However,the process of circular aptamer selection remains a great challenge,as it requires multiple rounds of binding-separation-amplification that involves issues with nonspecific binding and amplification bias.Here,we develop a highly practical solution for reliable selection of circular aptamers in a single round based on magnetosome-like magnetic chain cross-linked graphene oxide(separation efficiency≈105).High-affinity aptamer candidates can be rapidly selected from a preenriched circular DNA library,while low-affinity candidates are effectively adsorbed and separated by magnetosome-like magnetic chain cross-linked graphene oxide.With lipopolysaccharide as a representative model,the single-round selected lipopolysaccharide circular aptamer has been identified to have a high binding affinity with a Kd value of low to nanomolar range.Using this method,circular aptamers for protein and small-molecule targets were also successfully generated.We envision that this approach will accelerate the discovery of various new circular aptamers and open up a new avenue for analytical and therapeutic studies.展开更多
Biosynthesis of the functional factor𝛾γ-aminobutyric acid(GABA)in bacteria involves two key proteins an intra-cellular glutamate decarboxylase(GadB)and a membrane-bound antiporter(GadC).Efficient co-expressio...Biosynthesis of the functional factor𝛾γ-aminobutyric acid(GABA)in bacteria involves two key proteins an intra-cellular glutamate decarboxylase(GadB)and a membrane-bound antiporter(GadC).Efficient co-expression of suitable GadB and GadC candidates is crucial for improving GABA productivity.In this study,gadBΔC11 of Lacti-plantibacillus plantarum and gadCΔC41 of Escherichia coli were inserted into the designed double promoter(P T7lac and P BAD)expression system.Then,E.coli Lemo21(DE3)was chosen as the host to minimize the toxic effects of GadCΔC41 overexpression.Furthermore,a green and high-efficiency GABA synthesis system using dormant engineered Lemo21(DE3)cells as biocatalysts was developed.The total GABA yield reached 829.08 g/L with a 98.7%conversion ratio within 13 h,when engineered E.coli Lemo21(DE3)cells were concentrated to an OD 600 of 20 and reused for three cycles in a 3 M L-glutamate solution at 37℃,which represented the highest GABA productivity ever reported.Overall,expanding the active pH ranges of GadB and GadC toward physiological pH and employing a tunable expression host for membrane-bound GadC production is a promising strategy for high-level GABA biosynthesis in E.coli.展开更多
基金supported by the Research Funds of the Center for Big Data and Population Health of IHM(grant number JKS2022015)the Key Scientific Research Fund of the Anhui Provincial Education Department(grant number2023AH050610)the Anhui Natural Science Foundation(grant number 1808085QH252)。
文摘In 2021,approximately 537 million people suffered from diabetes mellitus(DM)globally,and this figure will increase to approximately 783 million within the next quarter-century.The increasing burden of DM is a pressing global public health issue.Therefore,the early identification of high-risk groups and implementation of effective intervention measures is imperative.
基金supported by the National Key R&D Program of China(2017YFC1600603)the Funds for Huangshan Professorship of Hefei University of Technology(407-037019).
文摘Food safety is a major issue to public health and have attracted global attention.Fast,sensitive,and reliable detection methods for food hazardous substances is highly desirable.Aptamers which can bind to the target molecules with high affinity and specificity represent an attractive tool for the recognition of food hazardous substances,which play an important role in the development and application of new food safety detection technology.But current assays for characterizing small molecule-aptamer binding are limited by either the mass sensitivity or the size differentiation ability.Herein,we proposed a comprehensive method for assessing the dissociation equilibria of small molecule-aptamer,which is immobilized-free under ambient conditions.The design employs the Le Chatelier’s principle and could be used to effectively measure small molecule-aptamer interactions.ATP binding aptamer and anti-aflatoxin B1 aptamer were used as the model system to determine their affinity,in which their dissociation equilibria measurements are in excellent close to their previous work.Due to the simplicity and sensitivity of this new method,we believe that it could be recommended as an effective tool for characterizing small molecule-aptamer interactions and promote the further application of small molecular aptamer in food safety.
基金supported by the National Natural Science Foundation of China(22104028,32072306,and U23A20265).
文摘Circular aptamers are promising candidates for analytical and therapeutic applications due to their enhanced biological and structural stability.However,the process of circular aptamer selection remains a great challenge,as it requires multiple rounds of binding-separation-amplification that involves issues with nonspecific binding and amplification bias.Here,we develop a highly practical solution for reliable selection of circular aptamers in a single round based on magnetosome-like magnetic chain cross-linked graphene oxide(separation efficiency≈105).High-affinity aptamer candidates can be rapidly selected from a preenriched circular DNA library,while low-affinity candidates are effectively adsorbed and separated by magnetosome-like magnetic chain cross-linked graphene oxide.With lipopolysaccharide as a representative model,the single-round selected lipopolysaccharide circular aptamer has been identified to have a high binding affinity with a Kd value of low to nanomolar range.Using this method,circular aptamers for protein and small-molecule targets were also successfully generated.We envision that this approach will accelerate the discovery of various new circular aptamers and open up a new avenue for analytical and therapeutic studies.
基金This work was supported by Natural Science Foundation of Zhe-jiang Province(LY23B060001)Zhejiang Provincial Key R&D Pro-gram of China(2021C02049)+2 种基金China Postdoctoral Science Founda-tion(2020M671337)National Natural Science Foundation of China(31670804,31971372)Ningbo"Scientific and Technological In-novation 2025″Key Project(2020Z080,2020Z088).
文摘Biosynthesis of the functional factor𝛾γ-aminobutyric acid(GABA)in bacteria involves two key proteins an intra-cellular glutamate decarboxylase(GadB)and a membrane-bound antiporter(GadC).Efficient co-expression of suitable GadB and GadC candidates is crucial for improving GABA productivity.In this study,gadBΔC11 of Lacti-plantibacillus plantarum and gadCΔC41 of Escherichia coli were inserted into the designed double promoter(P T7lac and P BAD)expression system.Then,E.coli Lemo21(DE3)was chosen as the host to minimize the toxic effects of GadCΔC41 overexpression.Furthermore,a green and high-efficiency GABA synthesis system using dormant engineered Lemo21(DE3)cells as biocatalysts was developed.The total GABA yield reached 829.08 g/L with a 98.7%conversion ratio within 13 h,when engineered E.coli Lemo21(DE3)cells were concentrated to an OD 600 of 20 and reused for three cycles in a 3 M L-glutamate solution at 37℃,which represented the highest GABA productivity ever reported.Overall,expanding the active pH ranges of GadB and GadC toward physiological pH and employing a tunable expression host for membrane-bound GadC production is a promising strategy for high-level GABA biosynthesis in E.coli.
