Factors that influence the extracellular expression of streptavidin in Escherichia coli using a bacteriocin release protein

Abstract

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.

GAP promoter‐based fed‐batch production of highly bioactive core streptavidin by Pichia pastoris

“…Comparison to literature. In comparison to literature, the maximal concentration for the optimized process surpassed titers for E. coli (typically 1‐2.6 µM), but was achieved for a lower productivity, as this parameter is usually in the range of 65 to 340 nM h −1 for this host . The productivity was close to the p STY of 67.9 nM h −1 achieved for P. pastoris by Nogueira et al The concentration of SAV was considerably lower than the reference concentrations for the methanol‐based production processes for SAV by P. pastoris reported by Casteluber et al (71 µM) and Nogueira et al (11 µM), but was achieved in a shorter cultivation time (72 vs. 167 h and 162 h, respectively).…”

Extracellular recombinant protein production from Escherichia coli

“…In another study, Miksch et al (2008) reported optimization of promoter strength for the kil gene for the extracellular expression of streptavidin (SA). A strong stationary-phase promoter for kil expression resulted in better SA expression and its extracellular secretion than using weak promoters.…”

Efficient production of extracellular proteins with Escherichia coli by means of optimized coexpression of bacteriocin release proteins

“…However, BRP coexpression is critical with respect to cell viability as the disintegration of the outer membrane rapidly stops cell division (van der Wal et al, 1995). Stationary phase dependent expression was found to be an appropriate strategy to overcome this drawback of BRP-mediated secretory protein production with E. coli (Miksch et al, 1997a(Miksch et al, , 2008Beshay et al, 2007). As an alternative approach, the P BAD promoter of the E. coli araBAD operon (Englesberg et al, 1969) was used in this work to allow arabinose induced BRP expression with maximum adjustability.…”