Cloning and Characterization of a Gene Cluster for Cyclododecanone Oxidation in Rhodococcus ruber SC1

Abstract

Biological oxidation of cyclic ketones normally results in formation of the corresponding dicarboxylic acids, which are further metabolized in the cell. Rhodococcus ruber strain SC1 was isolated from an industrial wastewater bioreactor that was able to utilize cyclododecanone as the sole carbon source. A reverse genetic approach was used to isolate a 10-kb gene cluster containing all genes required for oxidative conversion of cyclododecanone to 1,12-dodecanedioic acid (DDDA). The genes required for cyclododecanone oxidation were only marginally similar to the analogous genes for cyclohexanone oxidation. The biochemical function of the enzymes encoded on the 10-kb gene cluster, the flavin monooxygenase, the lactone hydrolase, the alcohol dehydrogenase, and the aldehyde dehydrogenase, was determined in Escherichia coli based on the ability to convert cyclododecanone. Recombinant E. coli strains grown in the presence of cyclododecanone accumulated lauryl lactone, 12-hydroxylauric acid, and/or DDDA depending on the genes cloned. The cyclododecanone monooxygenase is a type 1 Baeyer-Villiger flavin monooxygenase (FAD as cofactor) and exhibited substrate specificity towards long-chain cyclic ketones (C 11 to C 15 ), which is different from the specificity of cyclohexanone monooxygenase favoring short-chain cyclic compounds (C 5 to C 7 ).

Isolation and Characterization of an Alcohol Dehydrogenase Gene from the Octylphenol Polyethoxylate DegraderPseudomonas putidaS-5

“…2B). Intriguingly, an operon comprising an alcohol dehydrogenase and an esterase gene from Rhodococcus ruber SC1 34) or an acyl-CoA synthetase gene from P. putida P1 32) have been reported. They contribute to the biodegradation of cyclododecanone and alkane respectively, but the structure of these operons differed from that of P. putida S-5.…”

Metagenomic analysis of mesopelagic Antarctic plankton reveals a novel deltaproteobacterial group

“…They are involved in the extracellular degradation of complex compounds containing ester bonds and cyclic structures to produce simpler molecules, which are subsequently incorporated and metabolized in the cell (e.g. cyclododecanone; Kostichka et al, 2001). This might also be the case for DeepAnt-32C6 ORF 1, since it contains two predicted strong transmembrane helices, suggesting that it is most probably membrane-bound.…”

Application of a thermostable Baeyer–Villiger monooxygenase for the synthesis of branched polyester precursors

“…insertion of one atom of oxygen at the most‐ or least‐substituted position, respectively). BVMOs display a broad substrate scope,4, 5 which include cyclic ketones with a heteroatom,6, 7 macrocyclic ketones,8, 9, 10, 11 and substituted derivatives of various ring size (for example, terpene‐based substrates12, 13). 14, 15 BVMOs are also especially interesting in the synthesis of branched lactones, from which branched aliphatic polyesters with low glass transition temperatures can be made 16, 17, 18, 19, 20, 21, 22, 23, 24, 25…”