ORIGINAL RESEARCH PAPER Production of a recombinant alkane hydroxylase (AlkB2) from Alcanivorax borkumensis Mandana Miri • Bijan Bambai • Fatemeh Tabandeh • Majid Sadeghizadeh • Nasrin Kamali Received: 21 September 2009 / Revised: 10 November 2009 / Accepted: 10 November 2009 / Published online: 2 December 2009 Ó Springer Science+Business Media B.V. 2009 Abstract Alcanivorax borkumensis is an oil-degrad- ing 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. borkumen- sis 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. Keywords Alcanivorax borkumensis Á Alkane hydroxylase Á Biotransformation Á Rubredoxin Introduction Chemical hydroxylation of hydrocarbons is a challenging reaction. The C–H bond is nonpolar and activation of this bond in the Ziegler process requires high energy and metal catalysts (Azapagic et al. 2003). Production of long-chain alcohols (fatty alcohols) is currently performed through chemical processes with high energy demands, low yield and environmental contamination. However, there are alternative bio- technological systems capable of converting alkanes to the corresponding alcohols under ambient conditions with high specificity and negligible environmental constrains. There is a growing list of obligatory hydro- carbonoclastic microorganisms (van Beilen et al. 2003). These microorganisms represent a potentially Electronic supplementary material The online version of this article (doi:10.1007/s10529-009-0177-0) contains supplementary material, which is available to authorized users. M. Miri Science and Research Branch of Tehran, Islamic Azad University, Tehran 1477893855, Iran B. Bambai (&) Faculty of Biological Sciences, Shahid Beheshti University, GC, Tehran 19839, Iran e-mail: B_Bambai@sbu.ac.ir B. Bambai Á F. Tabandeh National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran 14178, Iran M. Sadeghizadeh Genetics Department, Tarbiat Modares University, Tehran 14115, Iran N. Kamali Malek-Ashtar University of Technology, Tehran 83145, Iran 123 Biotechnol Lett (2010) 32:497–502 DOI 10.1007/s10529-009-0177-0