Factors that influence the extracellular expression of streptavidin in Escherichia coli using a bacteriocin release protein.
Aiming to increase production of recombinant streptavidin in Escherichia coli, the effect of different leader sequences, different promoter strengths of the bacteriocin release protein (kil), host strain and medium composition on the expression and secretion into the medium was investigated. Expression vectors containing an expression or secretion unit were constructed with different combinations of leader sequence for the streptavidin gene and promoters for the kil gene and streptavidin gene. Results showed that a high-level extracellular production of streptavidin could be accomplished with E. coli BL21(DE3) by using the leader sequence of the phoA gene, a strong stationary-phase promoter for the kil gene and supplementation of the medium by glycine. Using a stationary-phase promoter for the expression of streptavidin had a negative effect.
Miksch
Gerhard
G
Forschungszentrum für Medizintechnik und Biotechnologie, 99947, Bad Langensalza, Germany. gmi@fermtech.techfak.uni-bielefeld.de
Ryu
Stella
S
Risse
Joe Max
JM
Flaschel
Erwin
E
D001430:Bacteriocins; D003470:Culture Media / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D019809:Streptavidin / Q000235:genetics / Q000378:metabolism*
D001430:Bacteriocins; D003470:Culture Media; D029968:Escherichia coli Proteins; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; C069003:kil protein, E coli; D019809:Streptavidin