Chromatographic Analysis of Recombinant Lysostaphin Expressed in Escherichia coli

Abstract

Lysostaphin (EC.3.4.24.75) is an extracellular glycylglycine endopeptidase produced exclusively by Staphylococcus simulans biovar staphylolyticus (ATCC 1362, NRRL B-2628). The zinc-containing endopeptidase demonstrates specific and potent staphylolytic activity therefore shows great promise for the treatment of blood-borne and biofilm-associated staphylococcal infections. The gene encoding mature lysostaphin has therefore been cloned and expressed using Escherichia coli, the most widely used expression host for recombinant protein production. Lysostaphin (EC.3.4.24.75) is an extracellular glycylglycine endopeptidase produced exclusively by Staphylococcus simulans biovar staphylolyticus (ATCC 1362, NRRL B-2628). The zinc-containing endopeptidase demonstrates specific and potent staphylolytic activity therefore shows great promise for the treatment of blood-borne and biofilm-associated staphylococcal infections. The gene encoding mature lysostaphin has therefore been cloned and expressed using Escherichia coli, the most widely used expression host for recombinant protein production. Lysostaphin (EC.3.4.24.75) is an extracellular glycylglycine endopeptidase produced exclusively by Staphylococcus simulans biovar staphylolyticus (ATCC 1362, NRRL B-2628). The zinc-containing endopeptidase demonstrates specific and potent staphylolytic activity therefore shows great promise for the treatment of blood-borne and biofilm-associated staphylococcal infections. The gene encoding mature lysostaphin has therefore been cloned and expressed using Escherichia coli, the most widely used expression host for recombinant protein production. Overall these findings are of concern for the production of recombinant protein production using E. coli. However this research suggested that chromatography can be used to sensitively detect and monitor product heterogeneity, so that a more homogeneous product could be produced by careful adjustment of expression, harvest and formulation conditions.

Date of Award	31 Aug 2011
Original language	English
Awarding Institution	Northumbria University
Supervisor	Gary Black (Supervisor)