摘要
【目的】从高温油藏中发掘新的微生物种质资源。【方法】采用Hungate厌氧操作技术从大港油田采出水中分离到一株厌氧杆菌HL-3。通过生理生化特征比较和16S rRNA序列比对,确定HL-3的分类地位。【结果】菌株HL-3为严格厌氧的革兰氏阴性杆菌。生长温度范围40℃-75℃(最适温度60℃);pH范围5.0-8.0(最适pH6.5);NaCl浓度范围0%-3.2%(最适NaCl浓度0.25%)。能够利用葡萄糖、核糖、甘露糖、木糖、纤维二糖等多种碳水化合物,发酵葡萄糖的产物是乙醇、乙酸、CO2及少量丙酸和丁醇。菌株HL-3的(G+C)mol%含量为33.9%,与Thermoanaerobacter(嗜热厌氧杆菌属)中模式菌株T.uzonensis DSM18761T(EF530067)的16S rRNA序列相似性为98.8%,与T.sulfurigignens DSM17917T(AF234164)的相似度次之为98.1%。菌株能够耐受浓度较高的亚硫酸根(0.1mol/L)离子和浓度极高的硫代硫酸根(0.8mol/L)。当硫代硫酸根浓度高于0.075mol/L时,菌体内产生硫单质颗粒;同时,在培养血清瓶顶空中检测到硫化氢气体。菌株HL-3与T.uzonensis DSM18761T对硫代硫酸根和亚硫酸根的耐受程度有很大不同。菌株HL-3对硫代硫酸根和亚硫酸根耐受程度及对硫代硫酸根的代谢机制与T.sulfurigignens DSM17917T(AF234164)极为相似,但二者代谢葡萄糖的产物却极不相同。【结论】所以菌株HL-3可能是Thermoanaerobacter属中的一个新种,其确切分类地位还有待用DNA分子杂交[1]的技术手段做进一步的鉴定。
[Objective]To find new microbial resources from a high-temperature oil reservoir.[Methods]Strain HL-3 was isolated by Hungate Anaerobic Technique from oil reservoir water sampled from Dagang oilfield,China. Through physiological,biochemical and phylogenetic analysis,the strain HL-3 was classified. [Results ] Cells were Gram-positive. The temperature range for growth was 40℃ -75℃ （ optimum at 60℃ ） and the pH range was 5. 0 -8. 0 （optimum at 6. 5）. The isolate could grow in the presence of 0% -3. 2% NaCl （optimum at 0. 25% ）. Glucose,ribose,mannose,xylose and cellobiose could be metabolized. Metabolites of glucose were ethanol,acetate,CO2 and trace amount of propionate and butanol. The G ＋ C content of DNA was 33. 9mol%. Based on 16S rRNA studies,strain HL-3 was most close to T. uzonensis DSM 18761T（EF530067） with 98. 8% similarity and to T. sulfurigignens DSM 17917^T（AF234164） with the 98. 1% similarity. Strain HL-3 tolerated to high sulfite （0. 1mol /L） ions and extremely high concentration of thiosulfate （0. 8 mol /L）. When the concentration of thiosulfate was higher than 0. 075 mol /L,the cell would generate S element granular. The presence of H2S gas was detected inside of space at the top of serum bottle. Strain HL-3 together with T. uzonensis DSM 18761T differed greatly in toleration of thiosulfate and sulfite. The toleration of strain HL-3 to thiosulfate and sulfite was most close to T. sulfurigignens DSM 17917T （ AF234164 ）. In addition,strain HL-3 to metabolite thiosulfate and sulfite was also similar with T. sulfurigignens DSM 17917T （ AF234164）. However,it differs largely from both of them to metabolize glucose. [Conclusion] Therefore,strain HL-3 may be a new spieces of the Thermoanaerobacter,and the definitive classification positioning is still awaiting for further verified with the method of determination of whole-genome DNA-DNA similarity.
出处
《微生物学报》
CAS
CSCD
北大核心
2010年第11期1525-1531,共7页
Acta Microbiologica Sinica
基金
国家自然科学基金(40973059)~~
关键词
热厌氧细菌
鉴定
硫代硫酸根
亚硫酸根
thermophilic anaerobic bacterium
identification
thiosulfate
sulfite
作者简介
刘海昌（1985-），男，山东潍坊人，硕士研究生，从事厌氧微生物研究。E-mail：haiehangliu198510@163．com
通信作者。Tel／Fax：＋86-28-85258573；E-mail：zhanghuits@yahoo．com．cn
