菲降解菌的特性及其降解酶纯化研究  

Characteristics and Enzyme Purification of Burkholderia sp.AFF for Phenanthrene Biodegradation

作　　者：贾玉红[1] 曲媛媛[1] 周集体[1] 李昂[1] 艾芳芳[1] 

机构地区：[1]大连理工大学环境与生命学院,辽宁大连116024

出　　处：《环境科学与技术》2009年第5期21-25,共5页Environmental Science & Technology

基　　金：国家自然科学基金项目资助(50608011)

摘　　要：研究考察了一株伯克霍尔德氏菌(Burkholderia sp.AFF)的基本特性,并对降解过程中的邻苯二酚2,3-双加氧酶进行了初步分离纯化。菌株AFF对链霉素和四环素具有抗性;当培养基中NaCl的浓度>1.5%时,菌株的生长降解能力受到抑制。菌株AFF生长与降解菲的最佳条件:温度30℃,pH=7.0,摇床转速为150r/min,接种量10%。菌株AFF的细胞粗提物具有邻苯二酚2,3-双加氧酶活性,用阴离子层析交换柱对其进行纯化,对收集到的具有酶活的组分进行SDS-PAGE分析,结果表明目的蛋白分子量在31kDa和43kDa之间。经过一步纯化后,比酶活提高33%,蛋白的回收率为27.6%。Characteristics and enzyme purification of Burkholderia sp. AFF were investigated. Strain AFF was resistant to streptomycin and tetracycline, when concentration of NaCl was more than 1.5%, abilities of growth and biodegradation for strain AFF were inhibited. The optimal conditions for both growth and phenanthrene-degradation process were as temperature 30℃, pH 7, shaking speed 150r/min and inoculation 10%. The strain contained an inductive catechol 2, 3-dioxygenase （C2, 30）, which was purified to homogeneity from the cell extracts of strain AFF by anion-exchange column and detected by SDS-PAGE. After one step purification, the protein yield was about 27.6% and enzyme activity was increased by 33%.

关 键 词：生物降解 菲 伯克霍尔德氏菌 邻苯二酚2 3-双加氧酶 

分 类 号：X172[环境科学与工程—环境科学]