Takii Susumu, Hanada Satoshi, Hase Yumiko, Tamaki Hideyuki, Uyeno Yutaka, Sekiguchi Yuji, Matsuura Katsumi
Department of Biological Sciences, Graduate School of Science, Tokyo Metropolitan University, Minamiohsawa 1-1, Hachioji 192-0397, Japan.
Int J Syst Evol Microbiol. 2008 Oct;58(Pt 10):2433-8. doi: 10.1099/ijs.0.65750-0.
To obtain amino acid-utilizing sulfate reducers, enrichment culture was carried out with a medium containing Casamino acids and sulfate and inoculated with coastal marine sediment from the eutrophic Tokyo Bay, Japan. A sulfate reducer, designated strain C/L2(T), was isolated from the sulfide-producing enrichment culture after further enrichment with lactate and sulfate by means of the agar shake dilution method. Cells of strain C/L2(T) were vibrio-shaped, Gram-negative, motile rods (0.7-1.0 mum wide and 1.0-3.5 mum long) with single polar flagella. The optimum temperature for its growth was 37 degrees C, the optimum pH was around 7.5 and the optimum NaCl concentration was 20-25 g l(-1). Hydrogen, formate, lactate, pyruvate, fumarate, malate, succinate, ethanol, propanol, glycerol, glycine, alanine, serine, aspartate, Casamino acids, peptone and yeast extract were used as electron donors. Sulfate, sulfite and thiosulfate each served as an electron acceptor, but elemental sulfur, nitrate, fumarate, acrylate and 2,4,6-tribromophenol did not. Disproportionation of thiosulfate was not observed. Desulfoviridin, c-type cytochromes and catalase were present. The major respiratory quinone was MK-6(H(2)). The G+C content of the genomic DNA was 46.2 mol%. Comparisons based on 16S rRNA gene sequences and on dissimilatory sulfite reductase gene sequences clearly showed that strain C/L2(T) belonged to the genus Desulfovibrio: its closest relatives were the uncharacterized Desulfovibrio sp. strain TBP-1 (16S rRNA gene sequence similarity of 99.4 %) and Desulfovibrio acrylicus DSM 10141(T) (16S rRNA gene sequence similarity of 98.7 %). The level of DNA-DNA hybridization with Desulfovibrio acrylicus DSM 10141(T) was 10.3 %. On the basis of the data from this study and the physiological and phylogenetic differences that exist between the isolate and Desulfovibrio acrylicus, strain C/L2(T) represents a novel species of the genus Desulfovibrio, for which the name Desulfovibrio marinisediminis sp. nov. is proposed. The type strain is C/L2(T) (=NBRC [corrected] 101113(T)=JCM 14577(T)=DSM 17456(T)).
为了获得利用氨基酸的硫酸盐还原菌,使用含有酪蛋白氨基酸和硫酸盐的培养基,并接种来自日本富营养化东京湾的沿海海洋沉积物进行富集培养。通过琼脂振荡稀释法,在进一步用乳酸盐和硫酸盐富集后,从产生硫化物的富集培养物中分离出一株硫酸盐还原菌,命名为菌株C/L2(T)。菌株C/L2(T)的细胞呈弧状,革兰氏阴性,具运动性的杆状(宽0.7 - 1.0μm,长1.0 - 3.5μm),有单根极生鞭毛。其生长的最适温度为37℃,最适pH约为7.5,最适NaCl浓度为20 - 25 g l⁻¹。氢气、甲酸盐、乳酸盐、丙酮酸盐、富马酸盐、苹果酸盐、琥珀酸盐、乙醇、丙醇、甘油、甘氨酸、丙氨酸、丝氨酸、天冬氨酸、酪蛋白氨基酸、蛋白胨和酵母提取物用作电子供体。硫酸盐、亚硫酸盐和硫代硫酸盐各自用作电子受体,但元素硫、硝酸盐、富马酸盐、丙烯酸盐和2,4,6 - 三溴苯酚则不能。未观察到硫代硫酸盐的歧化反应。存在脱硫绿素、c型细胞色素和过氧化氢酶。主要呼吸醌为MK - 6(H₂)。基因组DNA的G + C含量为46.2 mol%。基于16S rRNA基因序列和异化亚硫酸盐还原酶基因序列的比较清楚地表明,菌株C/L2(T)属于脱硫弧菌属:其最接近的亲缘种是未鉴定的脱硫弧菌属菌株TBP - 1(16S rRNA基因序列相似性为99.4%)和丙烯酸脱硫弧菌DSM 10141(T)(16S rRNA基因序列相似性为98.7%)。与丙烯酸脱硫弧菌DSM 10141(T)的DNA - DNA杂交水平为10.3%。基于本研究的数据以及该分离株与丙烯酸脱硫弧菌之间存在的生理和系统发育差异,菌株C/L2(T)代表脱硫弧菌属的一个新物种,为此提出新名称海洋沉积物脱硫弧菌(Desulfovibrio marinisediminis sp. nov.)。模式菌株为C/L2(T)(=NBRC [已校正] 101113(T)=JCM 14577(T)=DSM 17456(T))。