Science and Research Branch of Tehran, Islamic Azad University, Tehran 1477893855, Iran.
Biotechnol Lett. 2010 Apr;32(4):497-502. doi: 10.1007/s10529-009-0177-0. Epub 2009 Dec 2.
Alcanivorax borkumensis is an oil-degrading marine bacterium. Its genome contains genes coding for three cytochrome P450s and two integral membrane alkane hydroxylases (AlkB1 & AlkB2), all assumed to perform hydroxylation of different linear or branched alkanes. Although, the sequence of alkB2 has been determined, the molecular characterization and the substrate specificity of AlkB2 require more precise investigation. In this study, AlkB2 from A. borkumensis SK2 was expressed in Escherichia coli to examine the functionality of AlkB2 as a hydroxylating enzyme. Furthermore, the activity of the enzyme in the presence of the accessory proteins, rubredoxin (RubA) and rubredoxin reductase (RubB), produced in E. coli BL21(DE3)plysS cells, was determined. Recombinant AlkB2 is produced in an active form and rubredoxin is the intermediate electron donor to AlkB2 and can replace AlkG function, when NADH is the prime electron donor.
鲍曼不动杆菌是一种能降解石油的海洋细菌。其基因组包含编码三种细胞色素 P450 和两种完整膜烷烃羟化酶（AlkB1 和 AlkB2）的基因，所有这些基因都被认为能对不同的直链或支链烷烃进行羟化。尽管 AlkB2 的序列已经确定，但 AlkB2 的分子特征和底物特异性需要更精确的研究。在这项研究中，从 A. borkumensis SK2 中表达了 AlkB2，以研究 AlkB2 作为羟化酶的功能。此外，还测定了在 E. coli BL21(DE3)plysS 细胞中产生的辅助蛋白（Rubredoxin，RubA 和 Rubredoxin reductase，RubB）存在时，该酶的活性。重组 AlkB2 以活性形式产生，并且 Rubredoxin 是 AlkB2 的中间电子供体，可以在 NADH 是主要电子供体时取代 AlkG 的功能。
