降解蒽嗜盐菌AD-3的筛选、降解特性及加氧酶基因的研究  

Isolation,Charcaterization of an Anthracene Degrading Bacterium Martelella sp. AD-3 and Cloning of Dioxygenase Gene

作　　者：崔长征[1] 冯天才[1] 于亚琦[2] 董婓[1] 杨昕梅[1] 冯耀宇[1] 刘勇弟[1] 林汉平[1] 

机构地区：[1]华东理工大学资源与环境工程学院,国家环境保护化工过程环境风险评价与控制重点实验室,上海200237 [2]国家知识产权局专利局专利审查协作北京中心,北京100081

出　　处：《环境科学》2012年第11期4062-4068,共7页Environmental Science

基　　金：环境模拟与污染控制国家重点联合实验室开放基金项目(11K02ESPCT);中央高校基本科研业务费专项资金项目(WB1114031);高等学校博士学科点专项科研基金项目(20110074130002)

摘　　要：蒽是典型的多环芳烃类环境污染物,属于美国EPA优先控制的16种多环芳烃类化合物,其在高盐环境下的生物降解备受关注.本研究从某石油污染的高盐土壤中成功筛选出了1株高效降解蒽的菌株,经过对其生理生化特征和16S rDNA序列分析,初步鉴定并命名该菌株为Martelella sp.AD-3.该菌株在0.1%～10%的盐度和6.0～10.0的pH范围内,均能够降解蒽.其生长和降解蒽的优化条件是:蒽初始浓度25 mg·L-1、温度30℃、pH值9.0和盐度3%,在优化条件下培养6 d,蒽的降解率可达到94.6%.根据已报道的双加氧酶α亚基的同源性设计简并引物,通过巢式PCR扩增获得双加氧酶基因的部分序列307 bp(GenBank:JF823991.1),与海杆菌属Marinobacter sp.NCE312(AF295033)菌株萘双加氧酶大亚基的部分氨基酸序列同源性最高为95%.Anthracene,among the 16 US EPA polycyclic aromatic hydrocarbons（PAHs）,is a typical low molecular weight environmental contaminant,which gains concern on its biodegradation under hypersaline condition.In this study,an anthracene-degrading bacterial strain was isolated from highly saline petroleum-contaminated soil.Based on its physiological,biochemical characteristics and 16S rDNA sequence analysis,the bacteria was preliminary identified and named as Martelella sp. AD-3.The strain was able to utilize anthracene as sole carbon source for growth and the degradation occurred under broad salinities（0.1% to 10%） and varying pHs（6.0 to 10.0）.The optimized degradation conditions were initial concentration 25 mg·L^-1,culture temperature 30℃,pH 9.0 and salinity 3%.And 94.6% of anthracene was degraded by strain AD-3 under the optimal conditions within 6 days.Degenerate primers design was performed with a reported dioxygenase α subunit homologous gene.A length of 307 bp fragment of the partial dioxygenase gene sequences（GenBank accession： JF823991.1） was amplified by nested PCR.The clones amino acid sequence from strain AD-3 showed 95% identity to that of the partial naphthalene dioxygenase large-subunit from Marinobacter sp. NCE312（AF295033）.The results lay a foundation for the further study of molecular mechanism involved in the PAHs biodegradation by strain AD-3.

关 键 词：中度嗜盐菌 多环芳烃 蒽 生物降解 双加氧酶 

分 类 号：X172[环境科学与工程—环境科学]