Nucleotide sequences and regulational analysis of genes involved in conversion of aniline to catechol in Pseudomonas putida UCC22(pTDN1)
Nucleotide sequences and regulational analysis of genes involved in conversion of aniline to catechol in Pseudomonas putida UCC22(pTDN1)
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DOI：
10.1128/jb.179.2.399-408.1997
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发表时间：
1997-01-01
影响因子：
3.2
通讯作者：
Saint, CP
中科院分区：
文献类型：
--
作者：
Fukumori, F;Saint, CP
从恶臭假单胞菌Mt-2(UCC22)的衍生物中分离到的芳香胺分解质粒pTDN1的9,233个碱基的HindIII片段赋予恶臭假单胞菌KT2442降解苯胺的能力。该片段编码六个相同方向排列的开放阅读框。它们的5‘上游区域是pTDN1的直接重复序列的一部分。1.8kb的重复序列的核苷酸序列与参与氯苯甲酸酯基因转座的已知序列IS1071(C.Nakatsu，J.Ng，R.Singh，N.Straus和C.Wyndham，proc.)相比仅有单碱基变化。Natl，Acad，Sci，USA，88：8312-8316,1991)，四个开放阅读框编码的蛋白质与其他芳香族化合物降解途径中发现的蛋白质有相当大的同源性，根据序列相似性，这些基因编码苯胺加氧酶的大小亚基(分别为tdnA1和tdnA2)、还原酶(TdnB)和LysR型调节基因(TdnR)。推测的大亚基有一个保守的[2Fe-2S]R Rieske类型的配基中心，可能参与氨基转移的两个基因tdnQ和tdnT位于推测的加氧酶基因的上游；tdnQ基因的产物与谷氨酰胺合成酶有大约30%的相似性；然而，携带tdnQ的pUC载体在没有谷氨酰胺的情况下不支持大肠杆菌glnA菌株的生长，TdnT具有在酰胺转移酶中保守的结构域。TdnQ、tdnA1、tdnA2、tdnB和tdnR基因是苯胺转化为邻苯二酚所必需的。
A 9,233-bp HindIII fragment of the aromatic amine catabolic plasmid pTDN1, isolated from a derivative of Pseudomonas putida mt-2 (UCC22), confers the ability to degrade aniline on P. putida KT2442. The fragment encodes six open reading frames which are arranged in the same direction. Their 5' upstream region is part of the direct-repeat sequence of pTDN1. Nucleotide sequence of 1.8 kb of the repeat sequence revealed only a single base pair change compared to the known sequence of IS1071 which is involved in the transposition of the chlorobenzoate genes (C. Nakatsu, J. Ng, R. Singh, N. Straus, and C. Wyndham, Proc. Natl, Acad, Sci, USA 88:8312-8316, 1991), Four open reading frames encode proteins with considerable homology to proteins found in other aromatic-compound degradation pathways, On the basis of sequence similarity, these genes are proposed to encode the large and small subunits of aniline oxygenase (tdnA1 and tdnA2, respectively), a reductase (tdnB), and a LysR-type regulatory gene (tdnR). The putative large subunit has a conserved [2Fe-2S]R Rieske-type ligand center, Two genes, tdnQ and tdnT, which may be involved in amino group transfer; are localized upstream of the putative oxygenase genes; The tdnQ gene product shares about 30% similarity with glutamine synthetases; however, a pUC-based plasmid carrying tdnQ did not support the growth of an Escherichia coli glnA strain in the absence of glutamine, TdnT possesses domains that are conserved among amidotransferases. The tdnQ, tdnA1, tdnA2, tdnB, and tdnR genes are essential for the conversion of aniline to catechol.