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Elen Aquino Perpetuo,Regina Célia Pereira Marques,Maria Anita Mendes,Wanessa Cristina de Lima,Carlos Frederico Martins Menck,Claudio Augusto Oller do Nascimento 한국생물공학회 2009 Biotechnology and Bioprocess Engineering Vol.14 No.6
In this work, the biodegradation mechanism of phenol and sub products (such as catechol and hydroquinone) in Chromobacterium coli was investigated by cloning and molecular characterization of a phenol monooxygenase gene in Escherichia coli. This gene (Cvmp) is very similar (74 and 59% of similarity and identity, respectively) to the ortholog from Ralstomia eutropha, bacteria capable of utilizing phenol as the sole carbon source. The phenol biodegradation ability of E.coli recombinant strains was tested by cell-growth in a minimal medium containing phenol as the sole source of carbon and release of intermediary metabolites (catechol and hydroquinone). Interestingly, during the growth of these strains on phenol, catechol, and hydroquinone accumulated transiently in the medium. These metabolites were further analyzed by HPLC. These results indicated that phenol can be initially orto or para hydroxylated to produce cathecol or hydroquinone, respectively, followed by meta-cleavage of aromatic rings. To verify this information, the metabolites obtained from HPLC were submitted to LC/MS to confirm their chemical structure, thereby indicating that the recombinant strains utilize two different routes simultaneously, leading to different ring-fission substrates for the metabolism of phenol In this work, the biodegradation mechanism of phenol and sub products (such as catechol and hydroquinone) in Chromobacterium coli was investigated by cloning and molecular characterization of a phenol monooxygenase gene in Escherichia coli. This gene (Cvmp) is very similar (74 and 59% of similarity and identity, respectively) to the ortholog from Ralstomia eutropha, bacteria capable of utilizing phenol as the sole carbon source. The phenol biodegradation ability of E.coli recombinant strains was tested by cell-growth in a minimal medium containing phenol as the sole source of carbon and release of intermediary metabolites (catechol and hydroquinone). Interestingly, during the growth of these strains on phenol, catechol, and hydroquinone accumulated transiently in the medium. These metabolites were further analyzed by HPLC. These results indicated that phenol can be initially orto or para hydroxylated to produce cathecol or hydroquinone, respectively, followed by meta-cleavage of aromatic rings. To verify this information, the metabolites obtained from HPLC were submitted to LC/MS to confirm their chemical structure, thereby indicating that the recombinant strains utilize two different routes simultaneously, leading to different ring-fission substrates for the metabolism of phenol