1. Academic Validation
  2. A procedure for renaturation and purification of the extracellular Serratia marcescens nuclease from genetically engineered Escherichia coli

A procedure for renaturation and purification of the extracellular Serratia marcescens nuclease from genetically engineered Escherichia coli

  • Protein Expr Purif. 1994 Feb;5(1):37-43. doi: 10.1006/prep.1994.1005.
P Friedhoff 1 O Gimadutdinow T Rüter W Wende C Urbanke H Thole A Pingoud
Affiliations

Affiliation

  • 1 Institut für Biochemie, Fachbereich Biologie, Justus-Liebig-Universität, Giessen, Germany.
Abstract

Overproduction of the extracellular Serratia marcescens nuclease in Escherichia coli results in aggregation and sequestration of a large amount of the protein in inclusion bodies. Only a relatively small amount is secreted into the medium from which it can be purified following established procedures. The cell-associated insoluble protein can be solubilized in 6 M urea after breaking up the cells by sonication. Renaturation is achieved by dilution or dialysis. Subsequent phosphocellulose chromatography yields a homogeneous protein preparation which is shown by a variety of biochemical and biophysical analyses to be indistinguishable from conventionally prepared material. The high yield (> 10 mg/500-ml culture) and the ease of preparation (2 to 3 days) make this an attractive alternative to previously described procedures.

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