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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 or as a food preservative. One of the most important factors limiting application of the enzyme in clinical or technological practice is the high costs of its production. In the present study we have determined the optimal conditions for lysostaphin production in 5 l batch bioreactor. The enzyme production was based on, constructed earlier in our laboratory, heterologous, E. coli expression system assigned as pBAD2Lys. An evident influence of both 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. The efficiency of production of about 13000 U/L have been achieved in the optimal conditions of the production process: low aeration (400 rpm of mechanical stirrer), pH = 6 and temperature of 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 over 80 000 units of lysostaphin were obtained from one (batch) bioreactor −3 L of culture of E. coli TOP10 F' transformed with pBAD2Lys plasmid. From our best knowledge, it is the most efficient method of production of recombinant lysostaphin in E. coli expression systems described to date.
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- Category:
- Articles
- Type:
- artykuł w czasopiśmie wyróżnionym w JCR
- Published in:
-
PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
no. 44,
edition 4,
pages 370 - 381,
ISSN: 1082-6068 - Language:
- English
- Publication year:
- 2014
- Bibliographic description:
- Szweda P., Gorczyca G., Filipkowski P., Zalewska M., Milewski S.: Efficient production of Staphylococcus simulans lysostaphin in a benchtop bioreactor by recombinant Escherichia coli.// PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY. -Vol. 44, iss. 4 (2014), s.370-381
- Verified by:
- Gdańsk University of Technology
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