Characterization of catechol- and chlorocatechol-degrading activity in the ortho-chlorinated benzoic acid-degrading Pseudomonas sp. CPE2 strain.
Pyrocatechase activity was studied in the Pseudomonas sp. CPE2 strain, which is capable of growing on 2-chlorobenzoic and 2,5-dichlorobenzoic acid, giving rise to catechol and 4-chlorocatechol, respectively, as intermediate metabolites. The CPE2 crude extract was found to metabolize both catechol and 4-chlorocatechol. Enzymatic as well as phenotypic studies performed both on this strain and on a mutant strain lacking the chlorocatechol-degrading genes were consistent with the presence of two catechol-cleaving enzymes, one active mainly against catechol (pyrocatechase I) and the other with broader substrate specificity (pyrocatechase II). The latter enzyme also appeared to be induced when CPE2 cells were grown on 2-chlorobenzoic acid, thus contributing to catechol metabolism, in addition to pyrocatechase I. Despite the presence of a large plasmid in CPE2 cells, the chlorocatechol-degrading genes, highly homologous to the clc operon, were located on the chromosome. The selection at relatively high frequency of mutant strains with altered growth capabilities and which lacked the chlorocatechol-degrading genes suggests a transposon-like character for these catabolic genes in the CPE2 strain.
Department of Applied Chemistry and Material Science, University of Bologna, Italy.
D
D
Di Gioia
F
F
Fava
F
F
Baldoni
L
L
Marchetti
D001673:Biodegradation, Environmental; D015139:Blotting, Southern; D050548:Catechol 1,2-Dioxygenase; D002396:Catechols / Q000378:metabolism*; D002723:Chlorobenzoates / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003227:Conjugation, Genetic; D004269:DNA, Bacterial / Q000737:chemistry; D049308:Dioxygenases; D004587:Electrophoresis, Agar Gel; D009154:Mutation; D010105:Oxygenases / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D011549:Pseudomonas / Q000235:genetics / Q000378:metabolism*; D013011:Sorbic Acid / Q000031:analogs & derivatives / Q000032:analysis; D013379:Substrate Specificity
D002396:Catechols; D002723:Chlorobenzoates; D004269:DNA, Bacterial; C005939:muconic acid; D010105:Oxygenases; D049308:Dioxygenases; D050548:Catechol 1,2-Dioxygenase; D013011:Sorbic Acid