CLONING, NUCLEOTIDE-SEQUENCE, AND EXPRESSION OF THE GENE ENCODING A NOVEL DIOXYGENASE INVOLVED IN METABOLISM OF CARBOXYDIPHENYL ETHERS IN PSEUDOMONAS PSEUDOALCALIGENES POB310
CLONING, NUCLEOTIDE-SEQUENCE, AND EXPRESSION OF THE GENE ENCODING A NOVEL DIOXYGENASE INVOLVED IN METABOLISM OF CARBOXYDIPHENYL ETHERS IN PSEUDOMONAS PSEUDOALCALIGENES POB310
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DOI：
10.1007/bf00262201
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发表时间：
1995-01-01
影响因子：
2.8
通讯作者：
DWYER, DF
中科院分区：
文献类型：
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作者：
DEHMEL, U;ENGESSER, KH;DWYER, DF
假产碱假单胞菌菌株POB 310降解3-和4-羧基二苯醚。最初的反应涉及角双氧化产生不稳定的半缩醛，自发衰变为苯酚和原儿茶酸。我们从一个不稳定的、可自我传递的质粒中克隆了一个含有编码起始双加氧酶基因的DNA片段。序列分析显示两个开放阅读框编码的蛋白质的推定分子量为46.3和33.6 kDa。推导的氨基酸序列与芳香环活化双加氧酶的加氧酶和还原酶亚基具有同源性，并含有与叶绿体样和Rieske型[2Fe 2S]簇结合的共有序列相同的区域，表明起始双加氧酶是IA类芳香环活化双加氧酶系统。在含有3-或4-羧基二苯醚或苯酚作为碳源的M9基本培养基中生长的细菌中诱导初始双加氧酶活性，表明该调节依赖于苯酚途径。在指数生长期开始时测量最大比活性。
Pseudomonas pseudoalcaligenes strain POB310 degrades 3- and 4-carboxydiphenyl ether. The initial reaction involves an angular dioxygenation yielding an unstable hemiacetal that spontaneously decays to phenol and protocatechuate. We cloned a DNA fragment containing the gene encoding the initial dioxygenase from an unstable, self-transmissible plasmid. Sequence analysis revealed two open reading frames encoding proteins with putative molecular masses of 46.3 and 33.6 kDa. The deduced amino acid sequences showed homologies to oxygenase and reductase subunits of aromatic ring-activating dioxygenases, and contained regions identical to consensus sequences that bind chloroplast-like and Rieske-type [2Fe2S] clusters, suggesting that the initial dioxygenase is a class IA aromatic ring-activating dioxygenase system. Initial dioxygenase activity was induced in bacteria grown in M9 minimal medium containing 3- or 4-carboxydiphenyl ether or phenol as carbon source, indicating that the regulation is dependent on the phenol pathway. The maximal specific activity was measured at the beginning of the exponential growth phase.