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- Title
Optimization of extracellular truncated staphylococcal protein A expression in Escherichia coli BL21 ( DE3).
- Authors
Rigi, Garshasb; Mohammadi, Samira Ghaed; Arjomand, Maryam Rezaei; Ahmadian, Gholamreza; Noghabi, Kambiz Akbari
- Abstract
Staphylococcal protein A ( Sp A) plays an important role in Staphylococcus aureus pathogenesis. The recombinant Sp A is also widely used in biotechnology to purify polyclonal and monoclonal immunoglobulin G antibodies. In this study, expression and secretion of a truncated form of Sp A containing five immunoglobulin-binding domains using its own native signal sequence were optimized in Escherichia coli. Optimization was carried out using response surface method ( RSM), making use of the interaction between five variables. The initial results revealed that the signal peptide from S. aureus was recognized in E. coli and the resulting Sp A was expressed and secreted into the medium. Compounds, such as glycine, affected the secretion of Sp A into the culture medium. The central composite design experiment showed that the optimum conditions for the maximum expression of recombinant truncated Sp A in E. coli included 10% (w/v) lactose, 1.77% (w/v) glycine, induction time of 11 H, an optical density (600) of 1.1, and a temperature of 33 °C. Optimization using RSM resulted in a fivefold increase in the secretion of Sp A. To date, this is the first study of its kind regarding the definite influence of glycine concentration and duration of the cultivation period on the secretion of Sp A.
- Subjects
STAPHYLOCOCCAL protein A; ESCHERICHIA coli; IMMUNOGLOBULIN G; STAPHYLOCOCCUS aureus infections; PROTEIN expression; RESPONSE surfaces (Statistics)
- Publication
Biotechnology & Applied Biochemistry, 2014, Vol 61, Issue 2, p217
- ISSN
0885-4513
- Publication type
Article
- DOI
10.1002/bab.1157