Analysis of recombinant protein toxicity in E. coli through a phage λ-based genetic screening system

Guerau Fernández, Andrea Vera, Antonio Villaverde, Miguel Ángel Martínez

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4 Citations (Scopus)

Abstract

The aspartic protease from the human immunodeficiency virus type 1 (HIV-1) is highly toxic to E. coli, thus impairing its yield in production processes. Proteolytic cleavage of essential cellular proteins is probably a major contributor to the bacteriocidal effect but this has not been proven. Through an adapted high-throughput λ-based screening system, we have analyzed a set of HIV-1 protease mutants with distinguishable catalytic properties and we show that inactive enzymes are as toxic to E. coli cells as the wild-type enzyme. Together with additional data from directed molecular evolution approaches, these results indicate that the toxicity of the viral protease is not linked to its proteolytic activity. Our study also reveals that the λ-based screening system is a robust new tool for the genetic analysis of highly toxic recombinant products in E. coli. © 2007 Springer Science+Business Media B.V.
Original languageEnglish
Pages (from-to)1381-1386
JournalBiotechnology Letters
Volume29
Issue number9
DOIs
Publication statusPublished - 1 Jan 2007

Keywords

  • E. coli
  • Genetic screening
  • HIV-1
  • Protease
  • Protein production
  • Toxicity

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