Phosphoribosyl anthranilate isomerase is involved in the isomerization of phosphoribosyl anthranilate to 1-(o-carboxyphenylamino)-1-deoxyribulose 5-phosphate. In the present study, trpFGt, a gene encoding phosphoribosyl anthranilate isomerase from Geobacillus thermopakistaniensis, was cloned and expressed in Escherichia coli. The gene product, TrpFGt, was produced in E. coli in soluble and active form. Molecular characterization revealed that recombinant TrpFGt was highly efficient and stable. The apparent Vmax and Km values were 480 μmol min-1 mg-1 and 1.15 μM, respectively. The half-life of the enzyme was 90 min at 60 °C. Apart from thermostability, TrpFGt was highly stable against protein denaturants such as urea. There was no significant change in activity even after treatment with 8 M urea. To the best of our knowledge, TrpFGt, is the most active and stable phosphoribosyl anthranilate isomerase characterized to date and this is the first characterization of TrpF from the genus Geobacillus.

中文翻译：

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高效和稳定的热巴氏芽孢杆菌磷酸核糖基邻氨基苯甲酸异构酶的分子表征。

邻氨基苯甲酸磷酸核糖基酯异构酶参与了邻氨基苯甲酸磷酸核糖酯向5-磷酸1-（邻-羧基苯基氨基）-1-脱氧核糖的异构化。在本研究中，克隆trpFGt基因，该基因编码来自热巴氏芽孢杆菌的磷酸核糖基邻氨基苯甲酸酯异构酶，并在大肠杆菌中表达。基因产物TrpFGt以可溶性和活性形式在大肠杆菌中生产。分子表征表明重组TrpFGt是高效和稳定的。表观Vmax和Km值分别为480μmolmin-1 mg-1和1.15μM。酶的半衰期在60°C时为90分钟。除热稳定性外，TrpFGt还对蛋白质变性剂（如尿素）高度稳定。即使用8 M尿素处理后，活性也没有显着变化。据我们所知TrpFGt，