Development of a homologous expression system for rubber oxygenase RoxA from Xanthomonas sp.

Aims:  Natural rubber (poly‐[cis‐1,4‐isoprene]) can be cleaved into 12‐oxo‐4,8‐dimethyltrideca‐4,8‐diene‐1‐al by rubber oxygenase A (RoxA) isolated from Xanthomonas sp. RoxA is a novel type of dihaem dioxygenase with unknown cleavage mechanism of the rubber carbon backbone. Analysis of mutant RoxA after mutagenesis could be a way to investigate the function of selected amino acids of RoxA during catalysis. Unfortunately, expression of functional RoxA in recombinant Escherichia coli or in recombinant γ‐Proteobacteria such as Pseudomonas putida was not possible in our hands. Therefore, expression of recombinant RoxA in the homologous host, Xanthomonas, was performed.

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