About The Fatty Acid Electrochemistry Of GPR120 Receptor Sensor Research

Obesity has become a modern epidemic disease,which seriously endangers human health,affects the quality of life,and adds a huge medical burden to society.As a metabolic disease caused by overnutrition,obesity will become the world's largest public health problem in the 21st century.In 2010,China's obese population has reached 325 million.This figure is likely to double in the next 20 years.Therefore,it is extremely urgent to solve the problem of obesity in contemporary people.Fatty acid receptors are currently becoming a new potential target for the treatment of metabolic and inflammatory diseases.We need to better understand the molecular mechanisms behind this regulation,which may eventually lead to restrictions on eating new strategies that are rich in fats and harmful substances in order to properly cope with obesity.Taste is a special feeling that food produces by stimulating the chemosensory system in the mammalian mouth.In addition to acid,salty,bitter,sweet,and fresh,human taste is also the sixth taste-fat.They represent the calorie content of food intake,potential toxic substances,amino acid content,mineral presence,bacterial spoilage,etc.A variety of food ingredients and a sensitive taste perception system have made us a"tasteful"world.The electrochemical receptor sensor is a complex measuring instrument that uses electrochemistry as a cyclist and uses bioactive components as sensitive components.On this basis,the experiment used GPR120 receptor to prepare electrochemical receptor sensor and detected the interaction between 14 fatty acids and its receptor GPR120.The FFAR receptor gene GPR120 was found by NCBI,and the recombinant plasmid was obtained using the eukaryotic expression vector pcDNA3.0.The recombinant plasmid was transfected into HEK-239T cells by lipofection,and the target protein was obtained using the HEK-239T cell expression system.Then,chitosan,nanogold sol,GPR120 protein solution,thiopurine-chitosan mixture,horseradish peroxidase-nanogold sol,GPR120protein solution are sequentially fixed on the surface of the glassy carbon electrode by self-assembly method.The surface of the glassy carbon electrode was finally blocked with BSA solution.Each process of assembly was characterized using cyclic voltammetry.Different concentrations of solutions of 14 fatty acids were detected by time-current method using an electrochemical workstation.The results showed that GPR120 and 14 fatty acids showed hyperbolic curves in a certain concentration range,and the hyperbolic model was used for linear fitting.The correlation coefficient(R~2)was above 0.95.Using the double reciprocal method(Lineweaver-Burk equation)for mapping and regression analysis,R~2 also reached 0.95 or higher,and further calculated the allosteric constant of the acceptor-ligand:the destructive constant Ka value of acetic acid was 2.87×10~-1717 mol/L;the allosteric constant Ka value of propionic acid is 3.34×10~-1717 mol/L;the metamorphic constant Ka of n-butyric acid is 2.73×10~-1515 mol/L;the value of the allosteric constant of crotonic acid is 7.76.×10~-1616 mol/L;the ka value of the allosteric constant of n-octanoic acid is 7.47×10~-1616 mol/L;the value of the allosteric constant of ortho-acid is 2.38×10~-1515 mol/L;the variable constant of dodecanoic acid is Ka The value is 1.19×10~-1515 mol/L;the metamorphic constant Ka value of palmitic acid is 6.87×10~-1515 mol/L;the metamorphic constant Ka value of oleic acid is 8.32×10~-1515 mol/L;the structuring of linoleic acid The constant Ka value is 3.80×10~-1515 mol/L;the metamorphic constant Ka value of?-linolenic acid is 1.89×10~-1414 mol/L;and the allosteric constant Ka value of arachidonic acid is5.98×10~-1515 mol/L;The metamorphic constant Ka of EPA was 1.78×10~-1313 mol/L;the Ka value of the allosteric constant of DHA was 1.31×10~-1313 mol/L.This sensor can simulate the signal transmission process in vivo to some extent,revealing the relationship between receptor and ligand interaction.Using the same sensor to measure the same sample 10times,the RSD was 4.73%,indicating that the sensor has good repeatability.Five sensors of different batches were measured with an RSD of 2.52%,indicating that the sensor has good reproducibility.The electrode was stored above the PBS solution at 4?.On day 10,it was88.12%of the initial response signal,indicating that the sensor has good stability.