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RoxA

Part BBa_K1819003 corresponds to the coding sequence of rubber oxygenase A (Rox A) from Xanthomonas sp. strain 35Y (Gene bank accession code: AGT20506.1). This 73 kDa protein is an extracellular enzyme that catalyzes oxidative C-C cleavage of poly(cis-1,4-isoprene) to a major product, 12-oxo-4,8-dimethyltrideca-4,8-diene-1-al (ODTD; C15 tri-isoprenoid), with specific activity measured as 0.3 mol min-1 per mg. Found not only in Xanthomonas sp. but also in other Gram-negative clearing zone formers, RoxA is structurally related to cytochrome c peroxidases and its function is associated to two covalently bound heme groups. Although recently characterized by Birke et al (1), the exact enzymatic mechanism of rubber degradation by RoxA has not yet been elucidated. Because Xanthomonas sp. RoxA is exported to the extracellular environment, its open reading frame contains a sequence that codes to a signal peptide flanking its 5’ terminus. We detected this region (amino acid residues from 1 to 20) using signal peptide predictor server (4) (colocar link - http://www.compgen.org/tools/PRED-TAT) and eliminated it to generate part BBa_K1819003 with the coding sequence of RoxA flanked by SacI and NdeI restriction sites. Such non-usual restriction sites may serve for cloning purposes that lead to expression of recombinant protein with no additional amino acid residue(s) between its coding sequence and the fused N-terminal tag. Part BBa_K1819003 was cloned in pSB1C3 plasmid, sequenced and submitted to Registry of Standard Biological Parts.

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