Rabe Kersten S, Kiko Kathrin, Niemeyer Christof M
Technische Universität Dortmund, Fachbereich Chemie, Biologisch-chemische Mikrostrukturtechnik, Dortmund, Germany.
Chembiochem. 2008 Feb 15;9(3):420-5. doi: 10.1002/cbic.200700450.
We report the cloning, expression, and purification of CYP119, a thermostable enzyme previously thought to derive from Sulfolobus solfataricus. Sequence analysis suggested that, in contrast to the conclusions of earlier studies, the enzyme stems from the closely related Sulfolobus acidocaldarius, and we were indeed able to clone the gene from the genomic DNA of this organism. For the first time, we report here on the peroxidase activity of this enzyme and the optimization of the associated reaction parameters. The optimized reaction conditions were then applied to the biocatalytic epoxidation of styrene. The values obtained for k(cat) (78.2+/-20.6 min(-1)) and K(M) (9.2+/-4.3 mM) indicated an approximately 100-fold increased catalytic activity over previously reported results.
我们报告了CYP119的克隆、表达及纯化，CYP119是一种之前认为源自嗜热栖热菌的耐热酶。序列分析表明，与早期研究结论相反，该酶源自亲缘关系密切的嗜酸热硫化叶菌，并且我们确实能够从该生物体的基因组DNA中克隆出该基因。我们首次在此报告该酶的过氧化物酶活性及相关反应参数的优化。然后将优化后的反应条件应用于苯乙烯的生物催化环氧化反应。所获得的催化常数（k(cat)）（78.2±20.6 min⁻¹）和米氏常数（K(M)）（9.2±4.3 mM）的值表明，其催化活性比之前报道的结果提高了约100倍。
