Indigo production by Escherichia coli carrying the phenol hydroxylase gene from Acinetobacter sp strain ST-550 in a water-organic solvent two-phase system
Indigo production by Escherichia coli carrying the phenol hydroxylase gene from Acinetobacter sp strain ST-550 in a water-organic solvent two-phase system
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DOI：
10.1007/s00253-002-1187-1
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发表时间：
2003-02-01
影响因子：
5
通讯作者：
Aono, R
中科院分区：
文献类型：
--
作者：
Doukyu, N;Toyoda, K;Aono, R
不动杆菌ST-550菌株在大量二苯甲烷和高水平吲哚存在下从吲哚产生靛蓝。菌株ST-550在两相培养体系中生长时，某些蛋白质含量显著增加。其中一个蛋白的N端氨基酸序列与A.醋酸钙NCB 18250。当ST-550以苯酚为唯一碳源生长时，靛蓝产生活性被强烈诱导。根据与A. mopKLMOP的同源性，克隆了编码多组分苯酚羟化酶的基因。醋酸钙NCB 18250。携带这种基因的大肠杆菌能从吲哚中产生靛蓝。E. coli JA 300及其携带羟化酶基因和NADH再生系统的抗环己烷突变株OST 3410在两相培养系统中生长以生产靛蓝。OST 3410重组体在二苯甲烷存在下产生52 μ g靛蓝ml（-1）培养基。该生产率比JA 300重组体高4.3倍。
Acinetobacter sp. strain ST-550 produces indigo from indole in the presence of a large volume of diphenylmethane and a high level of indole. Particular proteins increased remarkably in strain ST-550 grown in the two-phase culture system for indigo production. One of the proteins showed a N-terminal amino acid sequence that was identical to that of the largest subunit of phenol hydroxylase (MopN) from A. calcoaceticus NCB18250. The indigo-producing activity was strongly induced when ST-550 was grown with phenol as a sole carbon source. Genes coding for the multicomponent phenol hydroxylase were cloned, based on the homology with mopKLMOP from A. calcoaceticus NCB18250. Escherichia coli carrying the genes produced indigo from indole. E. coli JA300 and its cyclohexane-resistant mutant, OST3410, carrying the hydroxylase genes and the NADH regeneration system were grown in the two-phase culture system for indigo production. The OST3410 recombinant produced 52 mug indigo ml(-1) of medium in the presence of diphenylmethane. This productivity was 4.3-fold higher than that of the JA300 recombinant.