Development of a Chemically Defined Medium for the Production of Enterolysin A From Enterococcus Faecalis B9510

J Appl Microbiol. 2013 Apr;114(4):1092-102. doi: 10.1111/jam.12115. Epub 2013 Jan 29.

Abstract

Aim: 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.

Conclusions: 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / chemistry
  • Bacteriocins / biosynthesis*
  • Bacteriocins / isolation & purification
  • Bacteriological Techniques
  • Bioreactors
  • Culture Media / chemistry*
  • Enterococcus faecalis / metabolism*
  • Vitamin B Complex / chemistry

Substances

  • Amino Acids
  • Bacteriocins
  • Culture Media
  • enterolysin A, Enterococcus faecalis
  • Vitamin B Complex