Heterologous expression of cholesterol 24S-hydroxylase, CYP46, in E. coli

We have developed a method for the heterologous expression of CYP46 in E. coli by using recombinant DNA technology to create expression constructs and then optimizing bacterial culture conditions to maximize protein synthesis. The expression constructs for CYP46 were successfully created using the pTrcHis-TOPO system. Expression of CYP46 in E. coli was optimized with respect to the various problems arising from the heterologous expression of a eukaryotic protein in a prokaryotic host. Human full length, human deleted, and mouse deleted constructs were used to obtain CYP46. As a result of this work new research projects are now possible to elucidate the role of CYP46 in AD.