期刊论文

中文

中国生物工程杂志

1671-8135

2016

36

5

59-67

10.13523/j.cb.20160509

国家自然科学基金资助项目（31460027）；

本校为其他机构

目的:从Aeromonas sp. XJ-6中克隆双加氧酶基因,初步探索该酶的功能,为芳香烃化合物的生物降解提供基因资源。方法:PCR扩增双加氧酶基因dio6,并实现该基因在大肠杆菌(Escherichia coli)中的诱导表达。产物经Ni-NTA柱纯化后,通过薄层层析(TLC)和HPLC检测双加氧酶dio6对Tyr的降解效果,再结合LC-MS检测降解产物,并分析其可能的降解途径。结果:Aeromonas sp. XJ-6双加氧酶基因dio6大小为1 194bp;通过金属鳌合亲和层析(MCAC)纯化后dio6表达产物的大小为44.9kDa。双加氧酶dio6对Tyr具有较强的降解作用。TLC和HPLC检测表明,在60μl酶量和30℃反应温度等条件下,Tyr降解较快;Mg2+、Ca2+略微抑制酶促反应,...

Objective: To clone the dioxygenase dio6 gene from Aeromonas sp. XJ-6, and explore the function of dioxygenase dio6 to provide genetic resources for biodegradation of aromatic compounds. Methods: Dioxygenase dio6 gene from Aeromonas sp.XJ-6 genome was amplified by PCR and induciblly expressed in Escherichia coli BL21(DE3). The expression products of dio6 was purified by Ni-NTA.TLC and HPLC to detect the degradation effect of dioxygenase on Tyr, and the degradation products were also detected by LC-MS so as to analyze the possible degradation pa...