Protein release in recombinant Escherichia coli using bacteriocin release protein.
The effect of induction level of the bacteriocin release protein (BRP) on cell growth characteristics, protein expression, and protein release in a recombinant strain of Escherichia coli RR1 was investigated. Mitomycin C, the inducing agent, when added to the growth medium in moderate amounts (up to 200 ng/mL) was observed to enhance the release of periplasmic proteins from the cell to the fermentation broth substantially. The percentages of release of the proteins alpha-amylase and beta-lactamase were increased by factors of about 7 and 3, respectively, upon induction of the BRP. The percentage of alpha-amylase released into the broth increased from only about 5% to almost 50% with the aid of BRP. The cell growth curve and low extracellular activity of the cytoplasmic protein beta-galactosidase were indicative that cell lysis did not occur in an appreciable amount at a low induction level, with a mitomycin C concentration of less than 300 ng/mL.
Department of Chemical Engineering, Rice University, Houston, Texas 77251-1892.
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D001426:Bacterial Proteins / Q000494:pharmacology*; D004926:Escherichia coli / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism*; D029968:Escherichia coli Proteins; D005285:Fermentation; D015636:Magnesium Chloride / Q000494:pharmacology; D016685:Mitomycin / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism*; D000516:alpha-Amylases / Q000096:biosynthesis / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism; D001618:beta-Lactamases / Q000096:biosynthesis / Q000378:metabolism
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; C052368:cloacin DF13-encoded bacteriocin release protein, E coli; D015636:Magnesium Chloride; D016685:Mitomycin; D000516:alpha-Amylases; D001616:beta-Galactosidase; D001618:beta-Lactamases