Construction of hybrid biphenyl (bph) and toluene (tod) genes for functional analysis of aromatic ring dioxygenases
Construction of hybrid biphenyl (bph) and toluene (tod) genes for functional analysis of aromatic ring dioxygenases
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用于芳环双加氧酶功能分析的杂合联苯 (bph) 和甲苯 (tod) 基因的构建
DOI：
10.1016/0378-1119(94)90779-x
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发表时间：
1994
期刊：
影响因子：
3.5
通讯作者：
Furukawa Kensuke
中科院分区：
文献类型：
--
作者：
Hirose Jun;Suyama Akiko;Hayashida Shinsaku;Furukawa Kensuke
假产碱假单胞菌菌株KF 707中由基因簇bphA 1A 2A 3A 4编码的联苯（BP）双加氧酶（Dox）的多组分酶复合物[Taira等人，J.Biol.Chem.267（1992）4844-4858]和由基因簇todC 1C 2BAinP编码的甲苯Dox。恶臭菌株F1 [Zylstra等人，J. Biol. Chem. 264（1989）14940-14946]，显示出高同源性（约100%）。60%），尽管它们具有离散的底物特异性。我们通过替换bphandtod基因簇中末端Dox大小亚基之间的基因组件构建了杂合基因簇，并分析了新型杂合芳香环Dox的功能。表达杂合基因簇todC 1：：bphA 2A 3A 4、todC 1C 2：：bphA 3A 4和bphA 1：：todC 2：：bphA 3A 4的大肠杆菌细胞获得了将苯-甲苯及其衍生物转化为二氢二醇的能力，表明由TodC 1：：BphA 2和BphA 1：：组成的杂合末端Dox TodC 2形成与铁氧还蛋白（Fer）（BphA 3）和Fer还原酶（BphA 4）相关的功能活性多组分Dox。此外，杂合Dox（由TodCl：：BphA 2A 3A 4和TodClC 2：：BphA 3A 4组成）显示出与野生型甲苯Dox（TodClC 2BA）相当相似的宽底物特异性。另一方面，杂合Dox（BphA 1：：TodC 2：：BphA 3A 4）对相同化合物显示氧化活性，但氧化速率取决于底物。这些结果表明：（i）末端Dox的两个亚基在BP、苯及其衍生物的底物特异性中起关键作用;（ii）电子传递蛋白Fer和Fer还原酶在BP Dox和甲苯Dox复合物之间是可交换的。
Multicomponent enzyme complexes of biphenyl (BP) dioxygenase (Dox) encoded by the gene cluster,bphA1A2A3A4 inPseudomonas pseudoalcaligenesstrain KF707 [Taira et al., J. Biol. Chem. 267 (1992) 4844–4858] and toluene Dox encoded by the gene cluster,todC1C2BAinP. putidastrain F1 [Zylstra et al., J. Biol. Chem. 264 (1989) 14940–14946], show high homologies (approx. 60%) for the corresponding subunit component in spite of the fact that they have discrete substrate specificities. We constructed hybrid gene clusters by replacing the gene component (s) between the large and small subunits of terminal Dox in thebphandtodgene clusters, and analyzed the function of a novel hybrid aromatic ring Dox.Escherichia colicells expressing the hybrid gene clusters,todC1::bphA2A3A4, todC1C2::bphA3A4andbphA1::todC2::bphA3A4, gained the ability to convert benzene-toluene and their derivatives to the dihydrodiols, indicating that the hybrid terminal Dox composed of TodCI::BphA2 and BphA1::TodC2 forms a functionally active multicomponent Dox associated with ferredoxin (Fer) (BphA3) and Fer reductase (BphA4). Moreover, hybrid Dox (composed of TodCl::BphA2A3A4 and TodC1C2::BphA3A4) showed a wide substrate specificity rather similar to that of the wild-type toluene Dox (TodClC2BA). On the other hand, the hybrid Dox (BphA1::TodC2::BphA3A4) showed oxidative activities for the same compounds, but the rate of oxidation was dependent upon the substrate. These results suggest that (i) the two subunits of terminal Dox are critically involved in the substrate specificity for BP, benzene and their derivatives, and (ii) the electron transport proteins, Fer and Fer reductase, are exchangeable with one another between the BP Dox and toluene Dox complexes.
铁氧还蛋白TOL 的纯化和性质。
DOI：
--
发表时间：
1985
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Subramanian,V;Liu,TN;Yeh,WK;Serdar,CM;Wackett,LP;Gibson,DT
通讯作者：
Gibson,DT
影响因子：
3.5
作者：
M. Simon;T. Osslund;R. Saunders;B. Ensley;S. Suggs;Arlette Harcourt;Suen Wen-chen;Diana L. Cruder;D. Gibson;G. Zylstra
通讯作者：
M. Simon;T. Osslund;R. Saunders;B. Ensley;S. Suggs;Arlette Harcourt;Suen Wen-chen;Diana L. Cruder;D. Gibson;G. Zylstra
铜绿假单胞菌 lasA 基因的核苷酸序列和在大肠杆菌中的表达。
影响因子：
3.2
作者：
Schad,PA;Iglewski,BH
通讯作者：
Iglewski,BH