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1 Marine Biotechnology Research Center, Korea Ocean Research & Development Institute, PO Box 29, Ansan 425-600, Republic of Korea
2 Extremobiosphere Research Center, JAMSTEC, 2-15 Natsushima-cho, Yokosuka City, Kanagawa 237-0061, Japan
Correspondence
Sang-Jin Kim
s-jkim{at}kordi.re.kr
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ABSTRACT |
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7c, 16 : 17c and cyclopropane 17 : 0. The major respiratory quinone was ubiquinone 10. The DNA G+C content was 54.5 mol%. The isolate did not contain bacteriochlorophyll a. Optimal growth required the presence of 2 % (w/v) NaCl with either 0.18 % CaCl2 or 0.59 % MgCl2, at pH 6.5 and at 35 °C. On the basis of the evidence of this polyphasic taxonomic study, strain JCS350T should be classified in a novel genus and species in the family Erythrobacteraceae, for which the name Altererythrobacter epoxidivorans gen. nov., sp. nov. is proposed. The misclassified species [Erythrobacter] luteolus is transferred to the new genus as Altererythrobacter luteolus comb. nov. The type strain of Altererythrobacter epoxidivorans is JCS350T (=KCCM 42314T =JCM 13815T) and the type strain of Altererythrobacter luteolus is SW-109T (=KCTC 12311T =JCM 12599T).
The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain JCS350T is DQ304436.
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, includes the genera Erythrobacter, Erythromicrobium and Porphyrobacter. Members of this family are aerobic and chemo-organotrophic. Most members contain bacteriochlorophyll a (BChl a) and various types of carotenoids. Certain species do not contain BChl a, including Erythrobacter citreus, Erythrobacter flavus and Erythrobacter luteolus (Denner et al., 2002; Yoon et al., 2003, 2005; Lee et al., 2005). All members of the genus Erythrobacter were isolated from seawater environments, whereas members of the genus Erythromicrobium were isolated from freshwater environments and members of the genus Porphyrobacter from various aquatic environments (Hiraishi et al., 2002; Yoon et al., 2004; Hiraishi & Imhoff, 2005; Shiba & Imhoff, 2005; Yurkov, 2005b). Sly & Hugenholtz (2005) recommended that members of the family Erythrobacteraceae should be reclassified into a single genus because 16S rRNA gene sequence similarities between and within the genera are high; on the other hand, Lee et al. (2005) did not investigate all species belonging to the order Sphingomonadales, so the taxonomic positions of certain species were not clarified. In the present study, we report the results of a polyphasic taxonomic identification of the marine bacterial strain JCS350T. This strain was found to fall within a novel genus within the family Erythrobacteraceae that also incorporates the misclassified species [Erythrobacter] luteolus.
Strain JCS350T was isolated from marine sediments of a cold-seep area in Kagoshima Bay, Japan. About 1 g sediment was enriched with 50 p.p.m. each of pyrene and benzo[a]pyrene in a mineral liquid medium (composition is described by Kwon et al., 2005) for 2 weeks at 25 °C. Next, 100 µl slurry was serially diluted with pre-sterilized seawater and plated onto marine agar 2216 (MA; Difco). Among the colonies showing different morphologies that grew on MA after 2 weeks, a yellow-coloured colony was isolated and named JCS350T. The isolate was cultivated on MA for morphological and biochemical characterization.
Epoxide hydrolase activity was tested according to the procedure described elsewhere (Woo et al., 2007). Strain JCS350T displayed strong epoxide hydrolase activity (data not shown).
Unless otherwise stated, physiological and morphological characterization was performed as described elsewhere (Sohn et al., 2004; Kwon et al., 2005). Cellular pigments were extracted with methanol from culture grown on MA and their absorption spectra were measured with a spectrophotometer (UV-2410PC; Shimadzu). Growth under anaerobic conditions was determined after incubation in an anaerobic chamber (Forma) on MA. The physiological, biochemical and morphological characteristics of strain JCS350T are given in the genus and species descriptions and in Table 1.
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. The dominant fatty acid methyl esters in strain JCS350T were 18 : 17c (46.9 %), 16 : 17c (32.9 %), cyclopropane 17 : 0 (6.9 %), 16 : 0 (3.1 %) and 16 : 15c (2.2 %). This profile of dominant fatty acids was similar to that of [Erythrobacter] luteolus; however strain JCS350T did not contain noticeable amounts of hydroxyl fatty acids.
The major respiratory quinone was determined to be ubiquinone 10 (Q10) by HPLC analysis according to Collins (1985). The DNA G+C content was 54.5 mol%, as determined by HPLC using a reversed-phase Symmetry C18 column (Waters) (Stackebrandt & Liesack, 1993).
A phylogenetic analysis using the 16S rRNA gene sequence was conducted according to the procedure described by Kwon et al. (2005). The 16S rRNA gene sequence of the isolate was a continuous stretch of 1444 nt, and 1332 unambiguously aligned base pairs were aligned with all sequences of members of the family Erythrobacteraceae and several members of the family Sphingomonadaceae. Rhodospirillum rubrum ATCC 11170T (GenBank accession no. D30778) and Acetobacter aceti DSM 3508T (X74066) served as outgroups. The closest neighbour with a validly published name is [Erythrobacter] luteolus (95.9 % similarity to the type strain) (Yoon et al., 2005). Other members of the genus Erythrobacter had 16S rRNA gene sequence similarities between 94.6 and 95.4 % and similarities to members of the genus Porphyrobacter were in the range 94.5–95.2 %. However, phylogenetic analysis of 16S rRNA gene sequences from organisms with validly published names revealed that strain JCS350T does not share a phyletic line with the genera Erythrobacter, Erythromicrobium and Porphyrobacter (Fig. 1). Instead, the isolate produced a distinct phyletic line with [Erythrobacter] luteolus supported by a high bootstrap value.
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Another organism, ‘Citromicrobium bathyomarinum’, was affiliated to the family Erythrobacteraceae rather than the family Sphingomonadaceae in our analysis (Fig. 1
). It has already been noted that the genus ‘Citromicrobium’ has higher 16S rRNA gene sequence similarity to the genus Erythrobacter rather than the genus Sphingomonas (Yurkov, 2005a). However, it remains in the family Sphingomonadaceae because the genus was not reclassified when the family Erythrobacteraceae was separated from the family Sphingomonadaceae (Lee et al., 2005).
The results of phylogenetic analysis and similarity of 16S rRNA gene sequences among strain JCS350T, [Erythrobacter] luteolus SW-109T and other members of the family Erythrobacteraceae imply that isolate JCS350T and [Erythrobacter] luteolus SW-109T are not affiliated to existing genera of the family. Strain JCS350T and [Erythrobacter] luteolus share several characteristics such as the lack of BChl a, the failure to produce H2S, the absence of nitrate reductase and urease activities and the ability to hydrolyse Tween compounds; however, they display differences in the utilization patterns of carbohydrates, starch degradation and DNA G+C content, for example (Table 1). Consequently, strain JCS350T should be classified within a new genus and species in the family Erythrobacteraceae, for which the name Altererythrobacter epoxidivorans gen. nov., sp. nov. is proposed. Furthermore, the reclassification of [Erythrobacter] luteolus Yoon et al. 2005 as Altererythrobacter luteolus comb. nov. is proposed.
Description of Altererythrobacter gen. nov.
Altererythrobacter (Al.ter.e.ryth'ro.bac'ter. L. adj. alius, alterius another, other, different; N.L. masc. n. Erythrobacter a genus name; N.L. masc. n. Altererythrobacter another or different Erythrobacter, because the genus shows high similarity to the genus Erythrobacter but does not share its phyletic line).
Cells are Gram-negative and non-motile and the colour of cell suspensions and colonies is yellow. Produce oxidase and catalase. Do not contain BChl a as a photosynthetic pigment. Methanol-soluble pigment is characterized by absorption maxima at 447 and 473 nm. Require NaCl for growth. Anaerobic growth does not occur on MA or on MA supplemented with nitrate. The dominant fatty acid is 18 : 17c. Do not reduce nitrate. H2S is not produced. The DNA G+C content is 54.5–60.3 mol%. The dominant respiratory quinone is Q10. As determined by 16S rRNA gene sequence analysis, the genus is a member of the family Erythrobacteraceae, class Alphaproteobacteria. The type species is Altererythrobacter epoxidivorans.
Description of Altererythrobacter epoxidivorans sp. nov.
Altererythrobacter epoxidivorans (e.po.xi.di.vo'rans. N.L. n. epoxidum epoxide; L. part. adj. vorans devouring; N.L. part. adj. epoxidivorans epoxide-devouring).
In addition to the description of the genus, the following properties are displayed. Cells are pleomorphic and 0.42–0.58x0.58–0.63 (or 0.67–1.25) µm in size. Methanol-soluble pigment is characterized by absorption maxima at 310, 447 and 473 nm. Growth occurs between 20 and 40 °C (optimum 35 °C), at pH 6–8.5 (optimum pH 6.5) and in the presence of 0.5–9 % NaCl (optimum 2 %) with either 0.18 % (w/v) CaCl2 or 0.59 % (w/v) MgCl2. 
-Galactosidase activity is detected and assimilation of mannitol, N-acetylglucosamine, maltose and adipate is observed using the API 20 NE kit. Alkaline phosphatase, esterase, esterase lipase, leucine arylamidase, cystine arylamidase and trypsin activities are observed in the API ZYM kit. Degrades Tweens 40 and 80, D-trehalose, -hydroxybutyric acid, succinamic acid, L-alanyl glycine, L-glutamic acid, glycyl L-glutamic acid, L-proline, L-serine and DL-
-glycerol phosphate and weakly positive for 
-hydroxybutyric acid utilization on Biolog GN2 plates. Possesses epoxide hydrolase activity. The DNA G+C content of the type strain is 54.5 mol%. The dominant fatty acids are 18 : 17c (46.9 %), 16 : 17c (32.9 %), cyclopropane 17 : 0 (6.9 %), 16 : 0 (3.1 %) and 16 : 15c (2.2 %).
The type strain is JCS350T (=KCCM 42314T =JCM 13815T), isolated from cold-seep sediments of Kagoshima Bay, Japan.
Description of Altererythrobacter luteolus comb. nov.
Altererythrobacter luteolus (lu.te.o'lus. L. masc. adj. luteolus yellowish).
Basonym: Erythrobacter luteolus Yoon et al. 2005.
The description is identical to that given for Erythrobacter luteolus by Yoon et al. (2005). The type strain is SW-109T (=KCTC 12311T =JCM 12599T).
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ACKNOWLEDGEMENTS |
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REFERENCES |
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TOPABSTRACTMAIN TEXTREFERENCES |
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Denner, E. B. M., Vybiral, D., Koblí
ek, M., Kämpfer, P., Busse, H.-J. & Velimirov, B. (2002). Erythrobacter citreus sp. nov., a yellow-pigmented bacterium that lacks bacteriochlorophyll a, isolated from the western Mediterranean Sea. Int J Syst Evol Microbiol 52, 1655–1661.[Abstract]
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Sly, L. I. & Hugenholtz, P. (2005). Genus II. Blastomonas Sly and Cahill 1997, 567VP emend. Hiraishi, Kuraishi and Kawahara 2000a, 1117. In Bergey's Manual of Systematic Bacteriology, 2nd edn, vol. 2, part C, pp. 258–263. Edited by D. J. Brenner, N. R. Krieg, J. T. Staley & G. M. Garrity. New York: Springer.
Sohn, J. H., Kwon, K. K., Kang, J.-H., Jung, H.-B. & Kim, S.-J. (2004). Novosphingobium pentaromativorans sp. nov., a high-molecular-mass polycyclic aromatic hydrocarbon-degrading bacterium isolated from estuarine sediment. Int J Syst Evol Microbiol 54, 1483–1487.
Stackebrandt, E. & Liesack, W. (1993). Nucleic acids and classification. In Handbook of New Bacterial Systematics, pp. 158–160. Edited by M. Goodfellow & A. G. O'Donnell. London: Academic Press.
Woo, J. H., Hwang, Y. O., Kang, S. G., Lee, H. S., Cho, J. C. & Kim, S.-J. (2007). Cloning and characterization of three epoxide hydrolases from a marine bacterium, Erythrobacter litoralis HTCC2594. Appl Microbiol Biotechnol 76, 365–375.[CrossRef][Medline]
Yoon, J.-H., Kim, H., Kim, I.-G., Kang, K. H. & Park, Y.-H. (2003). Erythrobacter flavus sp. nov., a slight halophile from the East Sea in Korea. Int J Syst Evol Microbiol 53, 1169–1174.
Yoon, J.-H., Lee, M.-H. & Oh, T.-K. (2004). Porphyrobacter donghaensis sp. nov., isolated from sea water of the East Sea in Korea. Int J Syst Evol Microbiol 54, 2231–2235.
Yoon, J.-H., Kang, K. H., Yeo, S.-H. & Oh, T.-K. (2005). Erythrobacter luteolus sp. nov., isolated from a tidal flat of the Yellow Sea in Korea. Int J Syst Evol Microbiol 55, 1167–1170.
Yoon, J.-H., Kang, S.-J., Lee, M.-H., Oh, H. W. & Oh, T.-K. (2006). Porphyrobacter dokdonensis sp. nov., isolated from sea water. Int J Syst Evol Microbiol 56, 1079–1083.
Yurkov, V. V. (2005a). Genus III. "Citromicrobium" Yurkov, Krieger, Stackebrandt and Beatty 1999, 4523. In Bergey's Manual of Systematic Bacteriology, 2nd edn, vol. 2, part C, pp. 263–266. Edited by D. J. Brenner, N. R. Krieg, J. T. Staley & G. M. Garrity. New York: Springer.
Yurkov, V. V. (2005b). Genus V. Erythromicrobium Yurkov, Stackebrandt, Holmes, Fuerst, Hugenholtz, Golecki, Gad'on, Gorlenko, Kompantseva and Drews 1994c, 432VP. In Bergey's Manual of Systematic Bacteriology, 2nd edn, vol. 2, part C, pp. 268–273. Edited by D. J. Brenner, N. R. Krieg, J. T. Staley & G. M. Garrity. New York: Springer.
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