Assembly of native sized spider silk protein in Escherichia coli

Abstract

Naturally found spider silk and elastin protein attract attention. However, the structureand size of this protein limits expression in heterologous hosts, where the repetitivesequences in mRNA create secondary structures. And these structures decreaseribosome processivity and assist mRNA degradation. We solved these problems usingmetabolic engineering and increasing the particular tRNA pool, and native-size spidersilk protein produced increased titer. The results provide insight into approaches tocontrol expression of proteins containing high molecular weight and highly repetitivesequence. [This work was supported by the Technology Development Program to SolveClimate Changes on Systems Metabolic Engineering for Biorefineries; the Intelligent Synthetic Biology Center through the GlobalFrontier Project of the Ministry of Education, Science and Technology(MEST) through the National Research Foundation of Korea]