Enterococcus camelliae sp. nov., isolated from fermented tea leaves in Thailand

Abstract

A Gram-positive and catalase-negative coccus that formed chains, strain FP15-1 T , isolated from fermented tea leaves ('miang'), was studied systematically. The strain was facultatively anaerobic and produced L-lactic acid from glucose. Demethylmenaquinone (DMK-7) was the major menaquinone. Straight-chain unsaturated fatty acids C 16 : 1 and C 18 : 1 were the dominant components. The DNA G+C content was 37.8 mol%. On the basis of 16S rRNA and RNA polymerase a subunit (rpoA) gene sequence analysis, strain FP15-1 T was closely related toEnterococcus italicus KCTC 5373 T , with 99.2 and 93.8 % similarity, respectively. The strain could be clearly distinguished from E. italicus ATCC 5373T by low DNA-DNA relatedness (¡33.8 %)and phenotypic characteristics. Therefore, this strain represent a novel species of the genus Enterococcus, for which the name Enterococcus camelliae sp. nov. is proposed. The enterococci comprise an important group of lactic acid bacteria found ubiquitously in the environment, the gastrointestinal tract, traditional fermented foods and dairy products. The classification of the genus Enterococcus has undergone considerable changes as a consequence of the increasing number of species and also improvements in methods to discriminate between species (Baele et al., 2000;Merquior et al., 1994;Naser et al., 2005 (Fortina et al., 2004), by comparison of partial sequences for three housekeeping genes, phenylalanyl-tRNA synthase a subunit (pheS), RNA polymerase a subunit (rpoA) and the a subunit of ATP synthase (atpA), and as confirmed by DNA-DNA hybridization (Naser et al., 2006). In the present paper, we describe a novel Enterococcus strain from fermented tea leaves ('miang') based on phenotypic and chemotaxonomic characteristics, DNA-DNA relatedness and 16S rRNA and rpoA gene sequence analysis.Samples of fermented tea leaves were collected from Chiangmai province, in the northern part of Thailand. Cocci in chains were isolated from the samples using GYPCaCO 3 agar (Tanasupawat et al., 1992). GYP-sodium acetate-mineral salt broth (Tanasupawat et al., 1992) (pH 7.2) was used for working cultures. All tests were performed by incubating the cultures at 30 u C. Cell shape, size and arrangement and colony appearance were examined using cells grown on GYP agar for 3 days. Gram staining was done as described by Hucker & Conn (1923). Spore formation was examined in the Gram-stained specimen.Results of the oxidation/fermentation test and motility were examined in soft agar (Whittenbury, 1963). Catalase activity,The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA and rpoA gene sequences of strain FP15-1 T are respectively EF154454 and EF197993.16S rRNA gene sequence-based maximum-likelihood and maximumparsimony trees are available as supplementary material with the online version of this paper. hydrolysis of gelatin, aesculin, arginine and starch, nitrate reduction, production of gas from glucose, gluconate and citrate and acid formation from carbohydrates were tested as reported by Tanasupawat et al. (1992)...

Tannin-tolerant and Extracellular Tannase Producing Bacillus Isolated from Traditional Fermented Tea Leaves and Their Probiotic Functional Properties

“…Like in other fermented foods, the key microorganisms which play an important role in Miang fermentation are lactic acid bacteria [11][12][13]. In the last two decades, Lactobacillus pentosus, Lactobacillus vaccinostercus, Enterococcus casseliflavus, Enterococcus calmelliae, Lactobacillus thailandensis, Lactobacillus camelliae and Pediococcus siamensis have been reported to be involved in Miang fermentation [14,15]. Chaikaew et al [16] also reported that L. plantarum group was considered to be the predominant lactic acid bacteria in Miang.…”

“…Since tea leaves contain significant amounts of phenolic compounds, especially tannins that are known to be microbial growth inhibitors, the ability of microorganisms associated with the tea leaves fermentation to survive under tannin-rich conditions are of interest [17]. Many studies have evaluated tannin-tolerant lactic acid bacteria that have been isolated from Miang [11,[14][15][16]. Chaikaew et al [16] also confirmed that 23 isolates from 311 lactic acid bacterial strains isolated from Miang samples belonged to L. pentosus and exhibited high tannin-tolerant capabilities in medium containing 2.5% (w/v) tannins.…”

Antibiotic resistance of Enterococci isolated from raw camel milk in the South West of Algeria

“…These bacteria are widely distributed in a variety of environmental habitats, even when there is little or no input from human and/or animal fecal sources (Byappanahalli et al, 2012). In addition, they are considered as lactic acid bacteria found in traditional fermented foods and in the dairy products (Sukontasing et al, 2007).…”

Screening lactic acid bacteria with high yielding-acid capacity from pickled tea for their potential uses of inoculating to ferment tea products

“…nov. sp. et al (Okada et al 1986;Tanasupawat and Komagata 1995;Sukontasing et al 2007;Tanasupawat et al 2007;Klayraung and Okonogi 2009;Klayraung;and Okonogi 2009). The reason for this difference may be that the salt was added into pickled tea fermentation, and Miang tea is a non-salt fermented product (Okada et al 1986).…”

“…There are many reports on microbial fermented tea, but almost concentrated on the fermentation technology, health function and chemical components as well as isolation and identification of microbial strains (Okada et al 1986;Tanasupawat and Komagata 1995;Mo et al 2005;Jayabalan et al 2007;Sukontasing et al 2007;Tanasupawat et al 2007;Abe et al 2008;Jayabalan et al 2008;Mo et al 2008;Klayraung and Okonogi 2009;Klayraung;and Okonogi 2009; Highlights -The microbial species of pickled tea were much simple. -Lactobacillus plantarum is the dominant specie in pickled tea fermentation.…”