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Rhodococcus opacus  R7 is a Gram-positive bacterium isolated from a polycyclic aromatic hydrocarbon contaminated soil for its versatile metabolism; indeed the strain is able to grow on naphthalene,  o -xylene, and several long- and medium-chain  n -alkanes. In this work we determined the degradation of  n -alkanes in  Rhodococcus opacus  R7 in presence of  n- dodecane (C12),  n- hexadecane (C16),  n- eicosane (C20),  n- tetracosane (C24) and the metabolic pathway in presence of C12. The consumption rate of C12 was 88%, of C16 was 69%, of C20 was 51% and of C24 it was 78%. The decrement of the degradation rate seems to be correlated to the length of the aliphatic chain of these hydrocarbons. On the basis of the metabolic intermediates determined by the R7 growth on C12, our data indicated that  R. opacus  R7 metabolizes medium-chain  n -alkanes by the primary alcohol formation. This represents a difference in comparison with other  Rhodococcus  strains, in which a mixture of the two alcohols was observed. By GC-MSD analysis we also identified the monocarboxylic acid, confirming the terminal oxidation.  Moreover, the  alkB  gene cluster from  R. opacus  R7 was isolated and its involvement in the  n -alkane degradation system was investigated by the cloning of this genomic region into a shuttle-vector  E. coli - Rhodococcus  to evaluate the alkane hydroxylase activity. Our results showed an increased biodegradation of C12 in the recombinant strain  R. erythropolis  AP (pTipQT1- alk R7) in comparison with the wild type strain  R. erythropolis  AP. These data supported the involvement of the  alkB  gene cluster in the  n -alkane degradation in the R7 strain.

Biodegradation of variable-chain-length n-alkanes in Rhodococcus opacus R7 and the involvement of an alkane hydroxylase system in the metabolism