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首页> 外文期刊>Journal of bacteriology >A Streptomyces avermitilis gene encoding a 4-hydroxyphenylpyruvic acid dioxygenase-like protein that directs the production of homogentisic acid and an ochronotic pigment in Escherichia coli.
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A Streptomyces avermitilis gene encoding a 4-hydroxyphenylpyruvic acid dioxygenase-like protein that directs the production of homogentisic acid and an ochronotic pigment in Escherichia coli.

机译：阿维链霉菌基因编码4-羟苯基丙酮酸双加氧酶样蛋白，可指导大肠杆菌中高纯酸和计时色素的产生。

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摘要

A 1.5-kb genomic fragment isolated from Streptomyces avermitilis that directs the synthesis of a brown pigment in Escherichia coli was characterized. Since pigment production in recombinant E. coli was enhanced by the addition of tyrosine to the medium, it had been inferred that the cloned DNA might be associated with melanin biosynthesis. Hybridization studies, however, showed that the pigment gene isolated from S. avermitilis was unrelated to the Streptomyces antibioticus melC2 determinant, which is the prototype of melanin genes in Streptomyces spp. Sequence analysis of the 1.5-kb DNA that caused pigment production revealed a single open reading frame encoding a protein of 41.6 kDa (380 amino acids) that resembled several prokaryotic and eukaryotic 4-hydroxyphenylpyruvate dioxygenases (HPDs). When this open reading frame was overexpressed in E. coli, a protein of about 41 kDa was detected. This E. coli clone produced homogentisic acid (HGA), which is the expected product of the oxidation of 4-hydroxyphenylpyruvate catalyzed by an HPD, and also a brown pigment with characteristics similar to the pigment observed in the urine of alkaptonuric patients. Alkaptonuria is a genetic disease in which inability to metabolize HGA leads to increasing concentrations of this acid in urine, followed by oxidation and polymerization of HGA to an ochronotic pigment. Similarly, the production of ochronotic-like pigment in the recombinant E. coli clone overexpressing the S. avermitilis gene encoding HPD is likely to be due to the spontaneous oxidation and polymerization of the HGA accumulated in the medium by this clone.
机译：表征了从阿维链霉菌分离的1.5 kb基因组片段，该片段指导大肠杆菌中棕色色素的合成。由于向培养基中添加酪氨酸可增强重组大肠杆菌中色素的产生，因此可以推断出克隆的DNA可能与黑色素的生物合成有关。但是，杂交研究表明，从阿维链霉菌中分离出的色素基因与链霉菌的melC2决定簇无关，后者是链霉菌中黑色素基因的原型。对导致色素生成的1.5-kb DNA的序列分析揭示了一个单一的开放阅读框，该阅读框编码41.6 kDa（380个氨基酸）的蛋白质，类似于几种原核和真核4-羟基苯基丙酮酸双加氧酶（HPD）。当此开放阅读框在大肠杆菌中过表达时，检测到约41 kDa的蛋白质。该大肠杆菌克隆产生了高纯酸（HGA），该高纯酸是HPD催化的4-羟基苯基丙酮酸氧化的预期产物，并且还具有棕色色素，其特性类似于在链足尿症患者尿液中观察到的色素。碱尿症是一种遗传性疾病，其中无法代谢HGA导致尿液中这种酸的浓度增加，随后HGA氧化并聚合为计时色素。类似地，在过表达编码HPD的阿维链霉菌基因的重组大肠杆菌克隆中，类似卵圆形的色素的产生很可能是由于该克隆在培养基中积累的HGA的自发氧化和聚合所致。

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