Selection of Pseudomonas sp. strain HBP1 Prp for metabolism of 2-propylphenol and elucidation of the degradative pathway.
A mutant of Pseudomonas sp. strain HBP1, originally isolated on 2-hydroxybiphenyl, was selected for the ability to grow on 2-propylphenol as the sole carbon and energy source. In the mutant strain, which was designated as Pseudomonas sp. strain HBP1 Prp, the cellular induction mechanism involved in the synthesis of the NADH-dependent monooxygenase is changed. 2-Propylphenol, which is known to be a substrate of the monooxygenase, does not induce formation of the monooxygenase in the wild type but does have an induction effect in the mutant strain. Furthermore, in contrast to the wild type, mutant strain HBP1 Prp constitutively produces a small amount of monooxygenase and metapyrocatechase. The enzymes from strain HBP1 Prp catalyzing the first three steps in the degradation of 2-propylphenol--the NADH-dependent monooxygenase, the metapyrocatechase, and the meta fission product hydrolase--were partially purified, and their activities were measured. The product of the monooxygenase activity was identified by mass spectrometry as 3-propylcatechol. The metapyrocatechase used this compound as a substrate and produced a yellow meta fission product that was identified by mass spectrometry as 2-hydroxy-6-oxo-nona-2,4- dienoate. Butyrate could be detected as a product of the meta fission product hydrolase in crude cell extract of 2-propylphenol-grown cells, as well as an intermediate in culture broths during growth on 2-propylphenol. All three enzymes expressed highest activities for the metabolites of the degradation of 2-hydroxybiphenyl.
Kohler
H P
HP
Swiss Federal Institutes of Technology Zürich, Switzerland.
van der Maarel
M J
MJ
Kohler-Staub
D
D
D001673:Biodegradation, Environmental; D050557:Catechol 2,3-Dioxygenase; D002396:Catechols / Q000378:metabolism; D002849:Chromatography, Gas; D003470:Culture Media; D049308:Dioxygenases; D005231:Fatty Acids, Unsaturated / Q000096:biosynthesis; D006867:Hydrolases / Q000096:biosynthesis; D006899:Mixed Function Oxygenases / Q000096:biosynthesis; D010100:Oxygen / Q000378:metabolism; D010105:Oxygenases / Q000378:metabolism; D010636:Phenols / Q000378:metabolism*; D011549:Pseudomonas / Q000201:enzymology / Q000254:growth & development / Q000302:isolation & purification / Q000378:metabolism*
D002396:Catechols; D003470:Culture Media; D005231:Fatty Acids, Unsaturated; D010636:Phenols; C080640:2-hydroxy-6-oxo-nona-2,4-dienoate; C080638:2-propylphenol; D006899:Mixed Function Oxygenases; D010105:Oxygenases; D049308:Dioxygenases; D050557:Catechol 2,3-Dioxygenase; C080677:2-propylphenol monooxygenase; D006867:Hydrolases; C080639:3-propylcatechol; D010100:Oxygen