Six strains of a hitherto unknown, Gram-stain-positive coccus were recovered from samples of Spanish-style green-olive fermentations. The 16S rRNA gene sequences from these isolates shared 98.7 % and 98.5 % of their nucleotide positions with those from Enterococcus saccharolyticus subsp. taiwanensis 812T and from E. saccharolyticus subsp. saccharolyticus ATCC 43076T, respectively. The sequence of the rpoA gene in the isolates was 95 % similar to that of E. saccharolyticus CECT 4309T ( = ATCC 43076T). The 16S rRNA and rpoA gene phylogenies revealed that the isolates grouped in a statistically well-supported cluster separate from E. saccharolyticus. Enzyme activity profiles as well as fermentation patterns differentiated the novel bacteria from other members of the Enterococcusgenus. Finally, phenotypic, genotypic and phylogenetic data supported the identification of a novel species of the genus Enterococcus, for which the name Enterococcus olivae sp. nov. is proposed. The type strain is IGG16.11T ( = CECT 8063T = DSM 25431T).
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Funding
This study was supported by the:
Spanish Ministry of Science and Innovation
(Award AGL2009-07861)
Spanish Ministry of Economy and Competitiveness
(Award AGL2012-33400)
Junta de Andalucía Excellence
(Award AGR-07345 and AGR-04621)
AltschulS. F.,
MaddenT. L.,
SchäfferA. A.,
ZhangJ.,
ZhangZ.,
MillerW.,
LipmanD. J.(1997). Gapped blast and psi-blast: a new generation of protein database search programs. . Nucleic Acids Res25, 3389–3402. [View Article][PubMed]
CathcartD. P.(1995).Purification, characterization and molecular analysis of plantaricin S, a two-peptide bacteriocin from olive fermenting Lactobacillus plantarum strains. PhD thesis, Cranfield University;, Bedford, UK:.
CorsettiA.,
PerpetuiniG.,
SchironeM.,
TofaloR.,
SuzziG.(2012). Application of starter cultures to table olive fermentation: an overview on the experimental studies. . Front Microbiol3, 248. [View Article][PubMed]
De BellisP.,
ValerioF.,
SistoA.,
LonigroS. L.,
LavermicoccaP.(2010). Probiotic table olives: microbial populations adhering on olive surface in fermentation sets inoculated with the probiotic strain Lactobacillus paracasei IMPC2.1 in an industrial plant. . Int J Food Microbiol140, 6–13. [View Article][PubMed]
de CastroA.,
MontañoA.,
CasadoF.-J.,
SánchezA.-H.,
RejanoL.(2002). Utilization of Enterococcus casseliflavus and Lactobacillus pentosus as starter culture for Spanish-style green olive fermentation. . Food Microbiol19, 637–644. [View Article]
DevrieseL. A.,
PotB.(1995). The genus Enterococcus
. . In The Genera of Lactic Acid Bacteria, pp. 327–367. Edited by
WoodB. J. B.,
HolzapfelW. H.
. London:: Blackie Academic & Professional;. [View Article]
DevrieseL. A.,
CollinsM. D.,
WirthR.(1992). The genus Enterococcus
. . In The Prokaryotes: A Handbook on the Biology of Bacteria: Ecophysiology, Isolation, Identification, Applications, pp. 1465–1481. Edited by
BalowsA.,
TrüperH. G.,
DworkinM.,
HarderW.,
SchleiferK. H.
. New York:: Springer;.
DevrieseL. A.,
PotB.,
CollinsM. D.(1993). Phenotypic identification of the genus Enterococcus and differentiation of phylogenetically distinct enterococcal species and species groups. . J Appl Bacteriol75, 399–408. [View Article][PubMed]
FarrowJ. A. E.,
KruzeJ.,
PhillipsB. A.,
BramleyA. J.,
CollinsM. D.(1984). Taxonomic studies on Streptococcus bovis and Streptococcus equinus: description of Streptococcus alactolyticus sp. nov. and Streptococcus saccharolyticus sp. nov.. Syst Appl Microbiol5, 467–482. [View Article]
FranzC. M. A. D.,
HolzapfelW. H.,
StilesM. E.(1999). Enterococci at the crossroads of food safety?. Int J Food Microbiol47, 1–24. [View Article][PubMed]
GonzalezJ. M.,
Saiz-JimenezC.(2002). A fluorimetric method for the estimation of G+C mol% content in microorganisms by thermal denaturation temperature. . Environ Microbiol4, 770–773. [View Article][PubMed]
GonzalezJ. M.,
Saiz-JimenezC.(2005). A simple fluorimetric method for the estimation of DNA-DNA relatedness between closely related microorganisms by thermal denaturation temperatures. . Extremophiles9, 75–79. [View Article][PubMed]
KleinG.(2003). Taxonomy, ecology and antibiotic resistance of enterococci from food and the gastro-intestinal tract. . Int J Food Microbiol88, 123–131. [View Article][PubMed]
KullenM. J.,
Sanozky-DawesR. B.,
CrowellD. C.,
KlaenhammerT. R.(2000). Use of the DNA sequence of variable regions of the 16S rRNA gene for rapid and accurate identification of bacteria in the Lactobacillus acidophilus complex. . J Appl Microbiol89, 511–516. [View Article][PubMed]
LeeH. M.,
LeeY.(2008). A differential medium for lactic acid-producing bacteria in a mixed culture. . Lett Appl Microbiol46, 676–681. [View Article][PubMed]
NaserS. M.,
ThompsonF. L.,
HosteB.,
GeversD.,
DawyndtP.,
VancanneytM.,
SwingsJ.(2005). Application of multilocus sequence analysis (MLSA) for rapid identification of Enterococcus species based on rpoA and pheS genes. . Microbiology151, 2141–2150. [View Article][PubMed]
Ruiz-BarbaJ. L.,
MaldonadoA.,
Jiménez-DíazR.(2005). Small-scale total DNA extraction from bacteria and yeast for PCR applications. . Anal Biochem347, 333–335. [View Article][PubMed]
StackebrandtE.,
GoebelB. M.(1994). Taxonomic note: a place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. . Int J Syst Evol Microbiol44, 846–849. [View Article]
TeixeiraL. M.,
FacklamR. R.(2003).Enterococcus
. . In Manual of Clinical Microbiology, , 8th edn., pp. 422–433. Edited by
MurrayP. R.,
BaronE. J.,
JorgensenJ. H.,
PfallerM. A.,
YolkenR. H.
. Washington, DC:: American Society for Microbiology;.
WayneL. G.,
BrennerD. J.,
ColwellR. R.,
GrimontP. A. D.,
KandlerO.,
KrichevskyM. I.,
MooreL. H.,
MooreW. E. C.,
MurrayR. G. E.& other authors (1987). Report of the ad hoc committee on reconciliation of approaches to bacterial systematics. . Int J Syst Bacteriol37, 463–464. [View Article]