Cloning and Characterization of a Gene Cluster for Cyclododecanone Oxidation in Rhodococcus ruber SC1
| Autor: | Kristy N. Kostichka, Qiong Cheng, Stuart M. Thomas, Vasantha Nagarajan, Katharine J. Gibson |
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| Rok vydání: | 2001 |
| Předmět: |
Molecular Sequence Data
Genetics and Molecular Biology Flavin group Microbiology Cofactor Substrate Specificity chemistry.chemical_compound Hydrocarbons Alicyclic Gene cluster Escherichia coli Rhodococcus Amino Acid Sequence Cloning Molecular Molecular Biology Alcohol dehydrogenase Phenylacetone monooxygenase Flavin adenine dinucleotide Sequence Homology Amino Acid biology Chromosome Mapping Monooxygenase Cosmids biology.organism_classification Models Chemical Biochemistry chemistry Genes Bacterial Multigene Family Flavin-Adenine Dinucleotide Oxygenases biology.protein Oxidation-Reduction |
| Zdroj: | Journal of Bacteriology. 183:6478-6486 |
| ISSN: | 1098-5530 0021-9193 |
| DOI: | 10.1128/jb.183.21.6478-6486.2001 |
| Popis: | 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 ). |
| Databáze: | OpenAIRE |
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