Development of a chemically defined medium for the production of enterolysin A from Enterococcus faecalis B9510
Article first published online: 29 JAN 2013
© 2012 The Society for Applied Microbiology
Journal of Applied Microbiology
Volume 114, Issue 4, pages 1092–1102, April 2013
How to Cite
Khan, H., Flint, S.H. and Yu, P.-L. (2013), Development of a chemically defined medium for the production of enterolysin A from Enterococcus faecalis B9510. Journal of Applied Microbiology, 114: 1092–1102. doi: 10.1111/jam.12115
- Issue published online: 18 MAR 2013
- Article first published online: 29 JAN 2013
- Accepted manuscript online: 26 DEC 2012 10:27AM EST
- Manuscript Accepted: 15 DEC 2012
- Manuscript Revised: 2 DEC 2012
- Manuscript Received: 18 SEP 2012
- defined medium;
- Enterococcus faecalis ;
- enterolysin A;
- lactic acid bacteria
This study aimed to develop a simplified chemically defined medium that could sustain the growth and bacteriocin (enterolysin A) production by Enterococcus faecalis B9510.
Methods and Results
The nutritional requirements of E. faecalis B9510 in a chemically defined medium were determined by single omission experiments. It was observed that eight amino acids (arginine, glycine, histidine, isoleucine, leucine, methionine, tryptophan and valine), three B vitamins (nicotinic acid, Ca-pantothenic acid and pyridoxal) and magnesium sulphate were essential for growth. Based on this information, a Simplified Defined Medium (SDM) was formed consisting of 26 components. Comparison of SDM with M-17 showed that growth and bacteriocin production in SDM was similar to that in M-17. The bacteriocin from SDM was then purified by ultrafiltration. The retentate of ultrafiltration step was analysed by SDS-PAGE and the results showed a single active band in the gel, which was excised and analysed by mass spectrometry, which indicated that the active band was enterolysin A, a cell wall degrading bacteriocin.
A simplified defined medium can be formulated for the growth and bacteriocin production by Enterococcus faecalis, whose efficiency is comparable with that of a complex commercial medium.
Significance and Impact of the Study
The development of such a medium can be useful for bacteriocin production and subsequent purification in a simplified manner and, therefore, helpful in the identification of novel bacteriocins.