Cloning, expression and purification of extracellular serine protease Esp, a biofilm-degrading enzyme, from Staphylococcus epidermidis.
J Appl Microbiol
; 111(6): 1406-15, 2011 Dec.
Article
em En
| MEDLINE
| ID: mdl-21974778
ABSTRACT
AIMS:
Staphylococcus epidermidis Esp, an extracellular serine protease, inhibits Staphylococcus aureus biofilm formation and nasal colonization. To further expand the biotechnological applications of Esp, we developed a highly efficient and economic method for the purification of recombinant Esp based on a Brevibacillus choshinensis expression-secretion system. METHODS ANDRESULTS:
The esp gene was fused with the N-terminal Sec-dependent signal sequence of the B. choshinensis cell wall protein and a C-terminal hexa-histidine-tag gene. The recombinant Esp was expressed and secreted into the optimized medium as an immature form and subsequently activated by thermolysin. The mature Esp was easily purified by a single purification step using nickel affinity chromatography and showed proteolytic activity as well as Staph. aureus biofilm destruction activity.CONCLUSIONS:
The purification yield of the developed extracellular production system was 5 mg recombinant mature Esp per 20-ml culture, which was much higher than that of an intracellular production system in Escherichia coli (3 mg recombinant Esp per 1-l culture). SIGNIFICANCE AND IMPACT OF THE STUDY Our findings will be a powerful tool for the production and purification of recombinant Esp and also applicable to a large variety of recombinant proteins used for basic researches and biotechnological applications.
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Base de dados:
MEDLINE
Assunto principal:
Staphylococcus epidermidis
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Biofilmes
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Serina Proteases
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Brevibacillus
Idioma:
En
Ano de publicação:
2011
Tipo de documento:
Article