title: Isolation and characterization of genes encoding polycyclic aromatic hydrocarbon dioxygenase from acenaphthene and acenaphthylene degrading Sphingomonas sp. strain A4.
abstract: Sphingomonas sp. strain A4 is capable of utilizing acenaphthene and acenaphthylene as sole carbon and energy sources, but it is unable to grow on other polycyclic aromatic hydrocarbons (PAHs). The genes encoding terminal oxygenase components of ring-hydroxylating dioxygenase (arhA1 and arhA2) were isolated from this strain by means of the ability to oxidize indole to indigo of the Escherichia coli clone containing electron transport proteins from phenanthrene-degrading Sphingobium sp. strain P2. The translated products of arhA1 and arhA2 exhibited moderate sequence identity (less than 56%) to large and small subunits of dioxygenase of other ring-hydroxylating dioxygenases. Biotransformation with recombinant E. coli clone revealed the broad substrate specificity of this oxygenase toward several PAHs including acenaphthene, acenaphthylene, naphthalene, phenanthrene, anthracene and fluoranthene. Southern hybridization analysis revealed the presence of a putative arhA1 homologue on a locus different from that of the arhA1 gene. Insertion inactivation of the arhA1 gene in strain A4 suggested that the gene but not the putative homologue one was involved in the degradation of acenaphthene and acenaphthylene in this strain.
authors.affiliation: Biotechnology Research Center, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Onruthai
authors.initials: O
authors.lastname: Pinyakong
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Habe
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kouzuma
authors.forename: Hideaki
authors.initials: H
authors.lastname: Nojiri
authors.forename: Hisakazu
authors.initials: H
authors.lastname: Yamane
authors.forename: Toshio
authors.initials: T
authors.lastname: Omori
mesh_terms: D000073:Acenaphthenes / Q000378:metabolism; D001673:Biodegradation, Environmental; D003001:Cloning, Molecular; D049308:Dioxygenases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D011084:Polycyclic Aromatic Hydrocarbons / Q000737:chemistry / Q000378:metabolism*; D020578:Sphingomonas / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D000073:Acenaphthenes; D011084:Polycyclic Aromatic Hydrocarbons; C042553:acenaphthylene; D049308:Dioxygenases; C042552:acenaphthene