大肠杆菌中分泌表达.pdf

Biotechnology Advances 23 (2005) 177–202 /locate/biotechadv Research review paper Recombinant protein secretion in Escherichia coli F.J.M. Mergulha˜oa,*, D.K. Summersb, G.A. Monteiroa aCentro de Engenharia Biolo´gica e Quı´mica, Instituto Superior Te´cnico, Av. Rovisco Pais, Lisbon 1049-001, Portugal bDepartment of Genetics, University of Cambridge, Cambridge CB2 3EH, United Kingdom Received 25 August 2004; received in revised form 23 November 2004; accepted 30 November 2004 Available online 8 January 2005 Abstract The secretory production of recombinant proteins by the Gram-negative bacterium Escherichia coli has several advantages over intracellular production as inclusion bodies. In most cases, targeting protein to the periplasmic space or to the culture medium facilitates downstream processing, folding, and in vivo stability, enabling the production of soluble and biologically active proteins at a reduced process cost. This review presents several strategies that can be used for recombinant protein secretion in E. coli and discusses their advantages and limitations depending on the characteristics of the target protein to be produced. D 2004 Elsevier Inc. All rights reserved. Keywords: Recombinant proteins; Escherichia coli ; Secretion; Periplasm; Type II Contents 1. Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 178 2. Recombinant protein secretion . . . . . . . . . . . . . . . . . . . . . . . . . . . . 178 2.1. Type I secretion mechanism . . .