Enterocin TW21, a novel bacteriocin from dochi -isolated Enterococcus faecium D081821

Abstract

Aims: Purification and characterization of a novel bacteriocin produced by strain Enterococcus faecium D081821. Methods and Results: Enterococcus faecium D081821, isolated from the traditional Taiwanese fermented food dochi (fermented black beans), was previously found to produce a bacteriocin against Listeria monocytogenes and some Gram-positive bacteria. This bacteriocin, termed enterocin TW21, was purified from culture supernatant by ammonium sulfate precipitation, Sep-Pak C 18 cartridge, ion-exchange and gel filtration chromatography. Mass spectrometry analysis showed the mass of the peptide to be approximately 5300Á6 Da. The N-terminal amino acid sequencing yielded a partial sequence NH 2 -ATYYGNGVYxNTQK by Edman degradation, and it contains the consensus class IIa bacteriocin motif YGNGV in the N-terminal region. The open reading frame (ORF) encoding the bacteriocin was identified from the draft genome sequence of Enterococcus faecium D081821, and sequence analysis of this peptide indicated that enterocin TW21 is a novel bacteriocin. Conclusions: Enterococcus faecium D081821 produced a bacteriocin named enterocin TW21, the molecular weight and amino acid sequence both revealed it to be a novel bacteriocin. Significant and Impact of Study: A new member of class IIa bacteriocin was identified. This bacteriocin shows great inhibitory ability against L. monocytogenes and could be applied as a natural food preservative.

Genetic and Biochemical Evidence That Enterococcus faecalis Gr17 Produces a Novel and Sec-Dependent Bacteriocin, Enterocin Gr17

“…To determine the molecular mass of the mature peptide, enterocin Gr17 was purified by salt precipitation, cation exchange, gel filtration chromatography, and RP-HPLC. Different from other enterocins, such as enterocin RM6 (7145.0823 Da) (Huang et al, 2013), enterocin TW21 (5302.98 Da) (Chang et al, 2013), and enterocin AS-48 (7149 Da) (Abengózar et al, 2017), the molecular mass of purified enterocin Gr17 was 4,531.01 Da, which corresponded to a calculated molecular mass of 4,533.11 Da due to the formation of a disulfide bond. To some extent, the molecular mass result confirmed the novelty of enterocin Gr17.…”

“…Numerous enterocins from E. faecium have been reported. These include the class IIa enterocin NKR-5-3C from E. faecium NKR-5-3C (Himeno et al, 2012), class IIa enterocin TW21 from E. faecium D081821 (Chang et al, 2013), class IIc enterocin AS-48 (Abengózar et al, 2017), and class IId enterocin K1 (Ovchinnikov et al, 2017). Even though features of the various enterocins have been determined, genome information and details of the biosynthesis mechanisms have been unknown.…”

Purification, Characterization, Identification, and Anticancer Activity of a Circular Bacteriocin From Enterococcus thailandicus

“…To detect the lytic activity of the purified bacteriocins, a parallel zymogram TRIS-tricin-PAGE gel was prepared and run in a similar manner as previously described. Furthermore, after completion of the electrophoresis run, gel was placed onto Brain Heart Infusion (BHI) agar surface overlaid with the indicator strain (S. aureus ATCC 29213), then incubated overnight at 37 • C (Beukes et al, 2000;Chang et al, 2013). The gel was compared with the Coomassie stained one to locate the active band, and hence determining its approximate molecular weight.…”

Complete Genome Sequence of Bacteriocin-Producing Enterococcus faecium HY07

“…Several coding DNA sequences (CDSs) for the production of bacteriocins, namely, enterocin P (GenBank accession number AYA33186), which contains the consensus class IIa bacteriocin motif YGNGV in the N -terminal region, were found (11). The genes encoding bacteriocins (GenBank accession numbers AYA33274, AYA35563, and AYA35573) were found in the genome of E.…”