Extracellular production of Bacillus penicillinase by Escherichia coli carrying pEAP2
Biotechnology
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Summary
Escherichia coli HB 101 containing the plasmid pEAP2 with alkalophilic Bacillus no. 170 penicillinase gene was found to produce large amounts of penicillinase in LB-broth containing 0.2% glycerol. Extracellular production of the enzyme was not induced by lysis of the cells but was strongly stimulated by the addition of Na+, K+ or Li+, with about 90% of total activity being recovered in the culture fluid. Without the addition of these ions, however, more than 80% of the enzyme activity remained in the cellular fraction. The enzyme produced was very stable and no decrease in activity was observed after long cultivation.
Keywords
Enzyme Escherichia Coli Enzyme Activity Glycerol Bacillus
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References
- Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heineker HL, Boyer HW (1977) Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2:95–113Google Scholar
- Cornelis P, Digneffe C, Willemot K (1982) Cloning and expression of a Bacillus coaglans amylase gene in Escherichia coli. Mol Gen Genet 186:507–511Google Scholar
- Goldfarb DS, Rodriguez RL, Doi RH (1982) Translation block to expression of the Escherichia coli Tn9-derived chloramphenicol-resistance gene in Bacillus subtilis. Proc Natl Acad Sci USA 79:5886–5890Google Scholar
- Horikoshi K, Akiba T (1982) Alkalophilic Microorganisms. Springer, Berlin Heidelberg New YorkGoogle Scholar
- Imanaka T, Tanaka T, Tsunekawa H, Aiba S (1981) Cloning of the genes for penicillinase, pen P and pen I, of Bacillus licheniformis in some vector plasmids and their expression in Escherichia coli, Bacillus subtilis and Bacillus licheniformis. J Bacteriol 147:776–786Google Scholar
- Sargent MG (1968) Rapid fixed-time assay for penicillinase. J Bacteriol 95:1493–1494Google Scholar
- Sawai T, Takahashi I, Yamagishi S (1978) Iodometric assay method for beta-lactamase with various beta-lactam antibiotics as substrates. Antimicrob Agents Chemother 13:910–913Google Scholar
- Sunaga T, Akiba T, Horikoshi K (1976) Production of penicillinase by an Alkalophilic Bacillus. Agric Biol Chem 40:1363–1367Google Scholar
- Whitaker JR (1963) Determination of molecular weights of proteins by gel filtration of Sephadex. Anal Chem 35:1950–1953Google Scholar
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© Springer-Verlag 1983