Centre for Bioengineering & Biotechnology, College of Chemical Engineering, China University of Petroleum (East China), Qingdao, 266580, PR China.
Centre for Bioengineering & Biotechnology, College of Chemical Engineering, China University of Petroleum (East China), Qingdao, 266580, PR China.
Protein Expr Purif. 2021 Dec;188:105964. doi: 10.1016/j.pep.2021.105964. Epub 2021 Aug 26.
The gene of catechol 1, 2-dioxygenase was identified and cloned from the genome of Oceanimonas marisflavi 102-Na3. The protein was expressed in Escherichia coli BL21 (DE3) and purified to homogeneity of a dimer with molecular mass of 69.2 kDa. The enzyme was highly stable in pH 6.0-9.5 and below 45 °C and exhibited the maximum activity at pH 8.0 and 30 °C. Being the first characterized intradiol dioxygenase from marine bacteria Oceanimonas sp., the enzyme showed catalytic activity for catechol, 3-methylcatechol, 4-methylcatechol, 3-chlorocatechol, 4-chlorocatechol and pyrogallol. For catechol, K and V were 11.2 μM and 13.4 U/mg of protein, respectively. The enzyme also showed resistance to most of the metal ions, surfactants and organic solvents, being a promising biocatalyst for biodegradation of aromatic compounds in complex environments.
从海洋盐单胞菌 102-Na3 的基因组中鉴定和克隆了儿茶酚 1,2-双加氧酶基因。该蛋白在大肠杆菌 BL21 (DE3) 中表达，并纯化为二聚体，分子量为 69.2 kDa。该酶在 pH 6.0-9.5 和 45°C 以下高度稳定，在 pH 8.0 和 30°C 时表现出最大活性。作为海洋细菌海洋单胞菌属中第一个被表征的邻二醇加氧酶，该酶对儿茶酚、3-甲基儿茶酚、4-甲基儿茶酚、3-氯儿茶酚、4-氯儿茶酚和焦儿茶酚具有催化活性。对于儿茶酚，K 和 V 分别为 11.2 μM 和 13.4 U/mg 蛋白。该酶还表现出对大多数金属离子、表面活性剂和有机溶剂的抗性，是在复杂环境中生物降解芳香族化合物的有前途的生物催化剂。
