Monitoring of the multiple bacteriocin production by Enterococcus faecium NKR-5-3 through a developed liquid chromatography and mass spectrometry-based quantification system

• Published in: Journal of bioscience and bioengineering Vol. 114; no. 5; pp. 490 - 496
• Main Authors: Perez, Rodney H., Himeno, Kohei, Ishibashi, Naoki, Masuda, Yoshimitsu, Zendo, Takeshi, Fujita, Koji, Wilaipun, Pongtep, Leelawatcharamas, Vichien, Nakayama, Jiro, Sonomoto, Kenji
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.11.2012; Elsevier
• Subjects: Anti-Bacterial Agents - analysis; Anti-Bacterial Agents - biosynthesis; Antimicrobial peptides; Bacteria; Bacteriocin quantification; Bacteriocins - analysis; Bacteriocins - biosynthesis; Biological and medical sciences; Biotechnology; Chromatography, Liquid - methods; Electrospray ionization-liquid chromatography and mass spectrometry (ESI-LC/MS); enterocins; Enterococcus faecium; Enterococcus faecium - metabolism; Fermentation; Fundamental and applied biological sciences. Psychology; Induction; Lactic acid bacteria; liquid chromatography; mass spectrometry; monitoring; Multiple bacteriocin; Peptides - isolation & purification; Spectrometry, Mass, Electrospray Ionization - methods; Temperature; Lactic acid bacteria; Electrospray ionization-liquid chromatography and mass spectrometry (ESI-LC/MS); Bacteriocin quantification; Induction; Multiple bacteriocin; Bacteriocin; Liquid chromatography; Electrospray; Streptococcaceae; Enterococcus faecium; Production; Bacteria; Micrococcales; Mass spectrometry; Monitoring; Quantitative analysis
• ISSN: 1389-1723; 1347-4421
• DOI: 10.1016/j.jbiosc.2012.06.003

Enterococcus faecium NKR-5-3 produces four antimicrobial peptides referred here as enterocins NKR-5-3A, B, C and D. A two-step electrospray ionization-liquid chromatography and mass spectrometry (ESI-LC/MS)-based quantification system was developed to monitor its multiple bacteriocin production profiles, which is essential in understanding the complex production regulation mechanism of strain NKR-5-3. The developed ESI-LC/MS-based quantification system can easily monitor the multiple bacteriocin production of this strain. Using the developed system, the production of enterocin NKR-5-3B was found to be not as variable as those of the other enterocins in different cultivation media. Production of enterocin NKR-5-3B was also found to have a wider optimum incubation temperature (20–30°C) than enterocins NKR-5-3A, C and D (25°C). Furthermore, at least 2 nM of the bacteriocin-like inducing peptide, enterocin NKR-5-3D, regulated the production of NKR-5-3 enterocins except enterocin NKR-5-3B. These findings taken together suggest that enterocin NKR-5-3B has an independent production regulation mechanism from the other NKR-5-3 enterocins. The developed system could effectively pin-point the production profiles of the multiple bacteriocins of E. faecium NKR-5-3 under different fermentation conditions.