三株高效石油降解红球菌Rhooccusspp生物学特性及相关基因.doc

PAGE 8 三株石油降解红球菌(Rhodococcus spp.)特性及相关基因分析* 杨 智1 陈吉祥1** 秦 波2 李彦林1 张庆芳1 赵 霞1 张婷1 1兰州理工大学石油化工学院 兰州 730050 2中国科学院兰州化学物理研究所 兰州 730000 摘 要 从石油污染土壤分离了3株能高效降解石油的细菌KB1、2182和JC3-47，在石油培养基中培养3d，3株菌对石油降解率分别为41.02%、32.26%和55.90%。经细菌形态学、生理生化鉴定和16S rDNA序列分析确定3株菌属于红球菌属(Rhodococcus spp)，其中KB1与红平红球菌 (Rhodococcus erythropolis)、2182与马红球菌 (Rhodococcus equi)、JC3-47与庆笙红球菌 (Rhodococcus qingshengii) 的16S rDNA序列相似性分别为100%，初步确定三株菌属于红平红球菌、马红球菌和庆笙红球菌。在温度10～50℃、pH 3～9、盐度为0～5.0 % 条件下菌株能很好生长，KB1和2182的最适生长温度为35℃，JC3-47最适温度为30℃，KB1和2182还可在pH 2 和 9.0 % 盐浓度的极端条件下生长。菌株能以十二烷、十八烷、苯、甲苯、二甲苯和萘为唯一碳源生长，其中KB1和JC3-47还能在芘的培养基生长。GC-MS分析发现3株菌对中链及长链烷烃都具有较强降解能力。3株菌都能产生表面活性剂，并对十六烷具有一定的粘附能力。利用特异性PCR扩增，在3株菌中均检测到烷烃单加氧酶、芳香烃双加氧酶和邻苯二酚双加氧酶基因，在KB1和2182中还检测出联苯双加氧酶基因。图6表3参39 关键词 石油降解；红球菌；鉴定；性质；降解基因 CLC X172 Characterization and catabolic gene detection of three oil-degrading Rhodococcus spp.* YANG Zhi1, CHEN Jixiang1**, QIN Bo2, LI Yanlin1, ZHANG Qingfang1, ZHAO Xia1 ZHANG Ting1 1School of Petrochemical Engineering, Lanzhou University of Technology, Lanzhou 730050, China 2Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou 730000, China Abstract Three oil-degrading bacteria strains named KB1, 2182 and JC3-47 were isolated from the oil-contaminated soil samples. The strains could use crude oil as sole carbon source with the degrading rates of 41.02%, 32.26% and 55.90% when they were cultured in minimal medium containing crude oil for 3 days. The bacteria were identified to belong to genus Rhodococcus by morphology, physiology, biochemical and 16S rDNA sequence analyses. The similarities of 16S rDNA sequences between KB1 and Rhodococcus erythropolis, 2182 and Rhodococcus equi, JC3-47 and Rhodococcus qingshengii were 100% respectively. KB1 was preliminarily identifies as Rhodococcus erythropolis, 2182 as Rhodococcus equi, JC3-47 as Rhodococcus qingshengii. They grew well at 10～50℃, with the initial pH of 3～9 and the NaCl concentration of 0～5%. The optimal temperatures for bacterial grow