p-Cymene pathway in Pseudomonas putida: ring cleavage of 2,3-dihydroxy-p-cumate and subsequent reactions.
It was confirmed that 2,3-dihydroxy-p-cumate is a substrate for ring cleavage in Pseudomonas putida PL-W after growth with p-cymene or p-cumate. This compound was oxidized to pyruvate, acetaldehyde, isobutyrate, and carbon dioxide by extracts of cells, and these products appear in equimolar amounts. The transient appearance of compounds and 2,3-dihydroxy-p-cumate to a yellow intermediate (lambda max, 345 nm) without decarboxylation. Extracts of the benzene nucleus; this is followed by decarboxylation to give the 393-nm species, which gives rise to isobutyrate, acetaldehyde, and pyruvate by the hydrolytic route of meta cleavage of catechols, via 4-hydroxy-2-oxovalerate. This was confirmed with a mutant of P. putida PL-RF-1 that was unable to grow with p-cymene (or p-cumate) but was able to oxidize both compounds AND 2,3-DIHYDROXY-P-CUMATE TO A YELLOW  INTERMEDIATE (LAMBDA MAX, 345 NM) WITHOUT DECARBOXYLATION. Extrats of P. putida PL-W (wild type) or a revertant of the mutant PL-RF-1 catalyzed the decarboxlation of the 345-nm intermediate with transient formation of the compound that absorbed at 393 nm. The substrate specificities of the 3,4-dioxygenative ring cleavage enzyme, and the decarboxylase were determined in crude extracts of P. putida PL-W and Pseudomonas fluorescens 007. It was conclude that 3,4-dioxygenative cleavage and decarboxylation are sequential enzyme-catalyzed reactions common to both P. putida and P. fluorescens for the oxidation of 2,3-dihydroxybenzoates. Unlike P. putida PL-W, which exclusively use the hydrolase branch, P. fluorescens 007 uses the dehydrogenase branch of the meta pathways that diverge after ring cleavage and later converge at oxoenate intermediates.
D001565:Benzoates / Q000378:metabolism; D002245:Carbon Dioxide / Q000378:metabolism; D002262:Carboxy-Lyases / Q000378:metabolism; D002396:Catechols / Q000378:metabolism; D003653:Decarboxylation; D062385:Hydroxybenzoates / Q000378:metabolism*; D010101:Oxygen Consumption; D011549:Pseudomonas / Q000201:enzymology / Q000378:metabolism*; D013729:Terpenes / Q000378:metabolism*
D001565:Benzoates; D002396:Catechols; D062385:Hydroxybenzoates; D013729:Terpenes; D002245:Carbon Dioxide; C009135:2,3-dihydroxybenzoic acid; D002262:Carboxy-Lyases