DsbA-DsbAmut fusion chaperon improved soluble expression of human trypsinogen-1 in Escherichia coli

摘要: A co-expressing system of DsbA-DsbAmut was suggested for the first time to enhance soluble expression human trypsin-1. As a control, leaderless DsbA chaperone also co-expressed with Vectors pET39b-trypsin and pET28a-DsbADsbAmut-trypsin above two fusion tag were constructed. The strain vector expressed protein DsbA-trypsin in form inclusion bodies. While E. coli BL21 (DE3) pET28a-DsbA-DsbAmut-trypsin, trypsin achieved. Under optimized conditions, fraction accounted about 49.43% total DsbA-DsbAmut-trypsin proteins crude supernatant. purification yield 4.15% by nickel chelating chromatography 3.3 mg activated purity 88.68% obtained from 1 L LB broth. To detect possible functions series chaperons protein, we analyzed primary three-dimensional structure proteins, mainly focusing on compatibleness between chaperons. results that (1) besides function periplasm, or DsbAmut may act as signal sequences-like leader targeted periplasm partly relieved pressure overexpression body formation, (2) there significant compared DsbA-trypsin, charge hydrophobic balance recombinant DsbA-DsbAmut-trypsin.