Effect Of Fusion Label Acyl Carrier Protein On Thermal Stability Of Exogenous Proteins

In recent years,with the continuous research and exploration of fusion label technology,more and more fusion protein labels have been applied in practice.Because the research background of E.coli is clear,compared with other expression systems,it has many advantages such as cost-effective,cost-saving,easy steps,and easy to be purified later.Therefore,in the process of biochemistry and molecular biology and biotechnology industrialization.It is very important.Since many foreign proteins are expressed in the E.coli expression system,they are easy to produce inclusion bodies or affect the function of the foreign protein itself,and suitable fusion tags to ensure efficient production,practical application and long-term preservation of the target protein,so the fusion label is utilized.Techniques for increasing the solubility,expression,thermal stability,and even activity of proteins have been widely used.The use of relatively small molecular weight and super acidic proteins as fusion labels is a new solution.Acyl Carrier Protein(ACP)is an important component of the organism's fatty acid synthase and plays an important role in other metabolic processes in the cell.ACPs in different organisms exist in different forms,but their structure and function are similar,that is,a small molecule protein with acidity.In this experiment,ACP was fused with TEV protease,Green Fluorescent Protein(GFP)and ?-Amylase(Amy).After high-efficiency induction,the solubility of different fusion proteins in the presence or absence of fusion-tag ACP was explored under the corresponding conditions.Expression,stability,and activity differences.At the same time,the effect of ACP on the stability of the foreign protein when it is linked to the N-terminus or C-terminus of the foreign protein is explored.The experimental results show that when ACP is used as a fusion tag,the solubility and expression of foreign proteins can be increased under certain conditions.In addition,ACP can significantly improve the stability of foreign proteins and increase the temperature by about 10?;When it is linked to the N-terminus or C-terminus of the foreign protein,the effect on stability is the same;after enzyme activity analysis,although the fusion tag ACP affects the activity of the foreign protein in a short time,the reaction time is long enough to contain ACP fusion proteins work more efficiently than fusion proteins that do not contain ACP.