多环芳烃降解菌的分离鉴定及其生理特性研究  

Study on isolation, identification and physiological characteristics of PAHs-degrading bacteria

作　　者：杨轩[1] 张威[2] 李师翁[1] 张满效[2,3] 刘光锈[2] 陈拓[2] 胡平[1] 伍修锟[2] 台喜生[2] 陈伟[1] 

机构地区：[1]兰州交通大学化学与生物工程学院,兰州730070 [2]中国科学院寒区旱区环境与工程研究所,兰州730000 [3]兰州石化职业技术学院,兰州730060

出　　处：《环境科学学报》2012年第5期1033-1040,共8页Acta Scientiae Circumstantiae

基　　金：国家自然科学基金项目(No.40971034;30800154);中国博士后科学基金项目(No.20080430794);甘肃省科技计划项目(No.1011FKCA114;1010RJZA009;2GS047-A52-012);甘肃省重大技术创新项目(No.[2004]197号;[2004]194号)~~

摘　　要：从兰州石化污水处理厂的污泥中分离出一株能分别以萘、蒽为唯一碳源生长的细菌(BDP01).通过菌株形态观察及16S rDNA序列比对,确定该菌株属于假单胞菌属的绿脓杆菌(Pseudomonas aeruginosa).菌株在LB培养基中48h内呈对数生长,48h到72h进入生长稳定期,72h后开始进入衰亡期;其最佳培养温度为30℃,最适生长pH为中性或弱碱性,该菌能在萘和蒽的质量浓度分别为100mg·L-1和50mg·L-1的无机盐培养基中良好生长,与已报道的其他微生物比较,BDP01在较短时间内具有较强的降解多环芳烃能力,1%的接种量15d内可降解89.64%的萘和77.21%的蒽.采用PCR和琼脂糖凝胶电泳技术检测到菌株基因组中有邻苯二酚2,3-双加氧酶(C23O)基因.测序结果与NCBI数据库发布的C23O基因序列相似度为92%.该酶在20～65℃以邻苯二酚为底物时,细胞裂解液中酶活力变化范围为13.10～540.86U.A strain, designated as BDP01, was isolated from the sludge of Lanzhou Sewage Disposal Plant. It utilized naphthalene and anthracene as the sole carbon sources for its growth. Based on the morphological characterizations and alignment analysis of the 16S rDNA sequence, BDP01 was identified as Pseudomonas aeruginosa. The growth curve of the strain was in logarithmic phase within 48 h, in stationary phase after 48 h to 72 h, and in decline phase after 72 h, in LB liquid medium. The strain grew well in inorganic salt liquid medium containing 100 mg · L-1 naphthalene or 50 mg · L-1 anthracene. The optimal growth temperature was 30 ℃ and the optimal growth pH value was 7~8, a neutral or alkalescent condition. The strain had a strong ability to biodegrade polycyclic aromatic hydrocarbon. In the present experiment, when the inoculum concentration of the strain was 1%, the degradation rates of naphthalene and anthracene were 89.64% and 77.21%, respectively, after cultured for 15 days. Furthermore, catechol 2, 3-dioxygenase gene sequence of the strain was detected using the PCR method and agarose gel electrophoresis. The gene sequencing and alignment analysis showed that catechol 2, 3-dioxygenase gene sequence of the strain had 92% similarity with the closest C23O gene sequence in the NCBI. With catechol as the substrate, the activity of catechol 2, 3-dioxygenase in the strain cellysis solution was 13.10~540.86 U at 20~65 ℃ reactive temperature.

关 键 词：污泥 萘 蒽 邻苯二酚2 3-双加氧酶 绿脓杆菌 

分 类 号：X172[环境科学与工程—环境科学]