菲降解菌的特性及其降解酶纯化研究

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 for Phenanthrene Biodegradation

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(C2,3O),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%.