Catechol 2,3-oxygenase production by genetically engineered Escherichia coli and its application to catechol determination
Catechol 2,3-oxygenase production by genetically engineered Escherichia coli and its application to catechol determination
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基因工程大肠杆菌产生儿茶酚2,3-加氧酶及其在儿茶酚测定中的应用
DOI：
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发表时间：
1991
影响因子：
4.1
通讯作者：
N. Shinohara
中科院分区：
文献类型：
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作者：
M. Fujita;T. Kamiya;M. Ike;Y. Kawagoshi;N. Shinohara
儿茶酚2,3-加氧酶是由大肠杆菌产生的，含有从苯酚降解恶臭假单胞菌BH中克隆的pheB基因的重组质粒pBH100，用于测定酒液中的儿茶酚。大肠杆菌 JM103 (pBH100) 和 C600 (pBH100) 的活性分别比恶臭假单胞菌 BH 高约 5 倍和 8.5 倍。使用重组体培养液制备的粗提物，可以定量测定磷酸盐缓冲液、合成污水以及苯酚、苯甲酸盐和水杨酸盐的混合物以及丙酮酸钠溶液中0.1至3.0μg/ml的儿茶酚。除儿茶酚外，还可以测定3-甲基儿茶酚、4-甲基儿茶酚和4-氯儿茶酚。粗提物经10%丙酮冻干后，-20℃保存100天，其氧化酶活性完全保持。
Catechol 2,3-oxygenase was produced by Escherichia coli, harbouring the recombinant plasmid pBH100 which contained the pheB gene cloned from phenol-degrading Pseudomonas putida BH, and was applied for the determination of catechol in the liquor. E. coli JM103 (pBH100) and C600 (pBH100) showed, respectively, about 5 and 8.5 times higher activities than that of P. putida BH. Using the crude extract prepared from the culture broth of the recombinant, catechol between 0.1 and 3.0 μg/ml could be determined quantitatively in phosphate buffer, synthetic sewage and in mixtures of phenol, benzoate and sallcylate, and also in sodium pyruvate solution. In addition to catechol, 3-methylcatechol, 4-methylcatechol and 4-chlorocatechol could be determined. Oxygenase activity of the crude extract was maintained completely during the 100-day storage at −20°C after being freeze-dried with 10% acelone.
质粒NAH7的基因nahH的核苷酸序列和表达以及与TOL pWWO的基因xylE的同源性。
影响因子：
3.5
作者：
Ghosal,D;You,IS;Gunsalus,IC
通讯作者：
Gunsalus,IC