Synthetic Biology and the Production from Brazilian Spider Silk Protein Masp2 in E.coli System

Abstract

Spider silk fibers have high strength, toughness and extensibility. This biomaterial has numerous potential applications in medicine and broader materials‐related needs. With synthetic biology it is possible to construct and express genes encoding recombinant spider silk proteins. These proteins are characterized by their highly repetitive specific structural motifs. However, production of spider silk in Escherichia coli can be difficult because the DNA can be unstable resulting in deletions, and the repetitive sequences can lead to transcription and translation errors. Here we show spidroin protein silk (≥50 KDa) production of the major ampullate silk, Masp2, from the Brazilian spider (Parawixia bistriata) in E. coli. A silk monomer gene was designed and a 16mer plasmid constructed using a “head to tail” cloning strategy. BL21(DE03) with or without a plasmid with copies of glycyl‐tRNA and glycyl‐tRNA synthetase were induced with IPTG. Samples was analysed by SDS‐PAGE. Both bacteria produce the protein. In other studies similar protein sizes have been produced from N. clavipes, with the size directly associated with improved fiber properties. The amino acid sequence of Masp2 from P. bistriata shows differences in composition. Different spider silks may result in different types of materials, which will provide new types of protein‐based biomaterials.Support: University of Brasilia, CNPq, CAPES, Embrapa Genetics Resources and Biotechnology, the NIH P41 Resource Center (EB002520), Tufts University.