Shuang Yu

Cloning and Expression of Catechol 2,3-dioxygenase from Achromobacter Xylosoxidans LHB21

Achromobacter xylosoxidans LHB21 isolated from a cold oil-contaminated site could utilize alkanes from pentadecane (C15) to dotriacontane (C32) as the sole carbon source and energy. The C23O gene was cloned and sequenced. Nucleotide sequence analysis of the C23O gene revealed a 924 base pairs open reading frame (ORF) initiating at ATG and terminating at TGA which predicted a protein of 35.16 kDa containing 307 amino acid residues. By comparison, the C23O of A. xylosoxidans LHB21 showed 44–100 % identity in amino acid sequence with other C23Os. The C23O gene was overexpressed in E. coli BL21(DE3) and the gene product was identified by SDS-PAGE and Western blotting.