title: Efficient production of Staphylococcus simulans lysostaphin in a benchtop bioreactor by recombinant Escherichia coli.
abstract: Lysostaphin is an enzyme with bactericidal activity against Staphylococcus aureus and other staphylococcal species. In spite of many advantages and promising results of preliminary research, the enzyme is still not widely used in medicine, veterinary medicine, or as a food preservative. One of the most important factors limiting application of the enzyme in clinical or technological practice is the high cost of its production. In this study we have determined the optimal conditions for lysostaphin production in a 5-L batch bioreactor. The enzyme production was based on a heterologous, Escherichia coli expression system designated as pBAD2Lys and constructed earlier in our laboratory. An evident influence of physicochemical conditions of the process (areation, pH and temperature) and composition of the growing media on the amount and activity of produced enzyme was noticed. Efficiency of production of about 13,000 U/L has been achieved in the optimal conditions of the production process: low aeration (400 rpm of mechanical stirrer), pH 6, and temperature 37°C in classical LB medium. Further, about twofold improvement in the production efficiency of the enzyme was achieved as a result of modification of composition of growing media. Finally, more than 80,000 units of lysostaphin were obtained from one (batch) bioreactor with 3 L of culture of E. coli TOP10F' transformed with pBAD2Lys plasmid. To the best of our knowledge, this is the most efficient method of production of recombinant lysostaphin in E. coli expression systems described to date.
authors: a Department of Pharmaceutical Technology and Biochemistry , Gdansk University of Technology , Gdansk , Poland.
authors: Piotr
authors: P
authors: Szweda
authors: Grzegorz
authors: G
authors: Gorczyca
authors: Pawel
authors: P
authors: Filipkowski
authors: Magdalena
authors: M
authors: Zalewska
authors: Slawomir
authors: S
authors: Milewski
mesh_terms: D000891:Anti-Infective Agents, Local / Q000378:metabolism* / Q000494:pharmacology; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005285:Fermentation; D006801:Humans; D008248:Lysostaphin / Q000378:metabolism* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D013203:Staphylococcal Infections / Q000188:drug therapy; D013210:Staphylococcus / Q000201:enzymology*; D013211:Staphylococcus aureus / Q000187:drug effects
chemical_list: D000891:Anti-Infective Agents, Local; D011994:Recombinant Proteins; D008248:Lysostaphin