title: Identification and expression of a Burkholderia pseudomallei collagenase in Escherichia coli.
abstract: Extracellular protein profiles were compared for broth-grown cultures of Burkholderia pseudomallei and its avirulent close relative Burkholderia thailandensis. A number of protein bands were present in the B. pseudomallei profile but absent or less abundant in the B. thailandensis profile. Four such prominent secreted proteins were identified by using N-terminal sequencing coupled to searches of the B. pseudomallei genome sequence database. The genes for two proteins with similarity to carbohydrate-binding proteins, and a further protein homologous to known bacterial collagenases, were present in both B. pseudomallei and B. thailandensis. The putative collagenase gene was cloned and expressed as a fusion protein in Escherichia coli. Cell lysates from Escherichia coli containing the recombinant protein exhibited detectable gelatinase and collagenase activities.
authors.affiliation: Department of Medical Microbiology and Genitourinary Medicine, University of Liverpool, Duncan Building, Daulby Street, Liverpool L69 3GA, United Kingdom.
authors.forename: Lucille
authors.initials: L
authors.lastname: Rainbow
authors.forename: Mark C
authors.initials: MC
authors.lastname: Wilkinson
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Sargent
authors.forename: C Anthony
authors.initials: CA
authors.lastname: Hart
authors.forename: Craig
authors.initials: C
authors.lastname: Winstanley
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000032:analysis; D016957:Burkholderia pseudomallei / Q000201:enzymology*; D017364:Collagenases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis*
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D017364:Collagenases