Comparison of Two Methods for Purification of Enterocin B, a Bacteriocin Produced by Enterococcus faecium W3

Date

2015-11-17

Author

Dundar, Halil
ATAKAY, MEHMET
Celikbicak, Omuer
SALİH, BEKİR
Bozoglu, Faruk

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This study aimed to compare two different approaches for the purification of enterocin B from Enterococcus faecium strain W3 based on the observation that the bacteriocin was found both in cell associated form and in culture supernatant. The first approach employed ammonium sulfate precipitation, cation-exchange chromatography, and sequential reverse-phase high-performance liquid chromatography. The latter approach exploited a pH-mediated cell adsorption-desorption method to extract cell-bound bacteriocin, and one run of reverse-phase chromatography. The first method resulted in purification of enterocin B with a recovery of 4% of the initial bacteriocin activity found in culture supernatant. MALDI-TOF MS analysis and de novo peptide sequencing of the purified bacteriocin confirmed that the active peptide was enterocin B. The second method achieved the purification of enterocin B with a higher recovery (16%) and enabled us to achieve pure bacteriocin within a shorter period of time by avoiding time consuming purification protocols. The purity and identity of the active peptide were confirmed again by matrix-assisted laser desorption/ionization time-of flight (MALDI-TOF) mass spectrometry (MS) analysis. Although both approaches were satisfactory to obtain a sufficient amount of enterocin B for use in MS and amino acid sequence analysis, the latter was proved to be applicable in large-scale and rapid purification of enterocin B.

Subject Keywords

Biotechnology, Biochemistry, General Medicine

URI

https://hdl.handle.net/11511/67964

Journal

PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY

DOI

https://doi.org/10.1080/10826068.2014.958165

Collections

Department of Biology, Article

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Citation Formats

H. Dundar, M. ATAKAY, O. Celikbicak, B. SALİH, and F. Bozoglu, “Comparison of Two Methods for Purification of Enterocin B, a Bacteriocin Produced by Enterococcus faecium W3,” PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY, pp. 796–809, 2015, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/67964.