PMID- 15866873 OWN - NLM STAT- MEDLINE DCOM- 20050912 LR - 20210209 IS - 0021-9258 (Print) IS - 0021-9258 (Linking) VI - 280 IP - 28 DP - 2005 Jul 15 TI - A two-component hydroxylase involved in the assimilation of 3-hydroxyphenyl acetate in Pseudomonas putida. PG - 26435-47 AB - The complete catabolic pathway involved in the assimilation of 3-hydroxyphenylacetic acid (3-OH-PhAc) in Pseudomonas putida U has been established. This pathway is integrated by the following: (i) a specific route (upper pathway), which catalyzes the conversion of 3-OH-PhAc into 2,5-dihydroxyphenylacetic acid (2,5-diOH-PhAc) (homogentisic acid, Hmg), and (ii) a central route (convergent route), which catalyzes the transformation of the Hmg generated from 3-OH-PhAc, l-Phe, and l-Tyr into fumarate and acetoacetate (HmgABC). Thus, in a first step the degradation of 3-OH-PhAc requires the uptake of 3-OH-PhAc by means of an active transport system that involves the participation of a permease (MhaC) together with phosphoenolpyruvate as the energy source. Once incorporated, 3-OH-PhAc is hydroxylated to 2,5-diOH-PhAc through an enzymatic reaction catalyzed by a novel two-component flavoprotein aromatic hydroxylase (MhaAB). The large component (MhaA, 62,719 Da) is a flavoprotein, and the small component (MhaB, 6,348 Da) is a coupling protein that is essential for the hydroxylation of 3-OH-PhAc to 2,5-diOH-PhAc. Sequence analyses and molecular biology studies revealed that homogentisic acid synthase (MhaAB) is different from the aromatic hydroxylases reported to date, accounting for its specific involvement in the catabolism of 3-OH-PhAc. Additionally, an ABC transport system (HmgDEFGHI) involved in the uptake of homogentisic acid and two regulatory elements (mhaSR and hmgR) have been identified. Furthermore, the cloning and the expression of some of the catabolic genes in different microbes presented them with the ability to synthesize Hmg (mhaAB) or allowed them to grow in chemically defined media containing 3-OH-PhAc as the sole carbon source (mhaAB and hmgABC). FAU - Arias-Barrau, Elsa AU - Arias-Barrau E AD - Departamento de Bioquimica y Biologia Molecular, Facultad de Veterinaria, Universidad de Leon, 24007 Leon, Spain. FAU - Sandoval, Angel AU - Sandoval A FAU - Naharro, German AU - Naharro G FAU - Olivera, Elias R AU - Olivera ER FAU - Luengo, Jose M AU - Luengo JM LA - eng SI - GENBANK/AY929299 SI - GENBANK/AY929300 SI - GENBANK/AY937229 PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20050502 PL - United States TA - J Biol Chem JT - The Journal of biological chemistry JID - 2985121R RN - 0 (3-hydroxyphenylacetate) RN - 0 (Culture Media) RN - 0 (Phenylacetates) RN - EC 1.- (Mixed Function Oxygenases) RN - NP8UE6VF08 (Homogentisic Acid) SB - IM MH - Amino Acid Motifs MH - Amino Acid Sequence MH - Base Sequence MH - Biological Transport MH - Catalysis MH - Culture Media/metabolism MH - Escherichia coli/metabolism MH - Homogentisic Acid/chemistry/*metabolism MH - Mixed Function Oxygenases/metabolism/*physiology MH - Models, Chemical MH - Models, Genetic MH - Molecular Sequence Data MH - Mutation MH - Phenylacetates/*chemistry MH - Plasmids/metabolism MH - Protein Transport MH - Pseudomonas fluorescens/metabolism MH - Pseudomonas putida/*metabolism MH - Time Factors EDAT- 2005/05/04 09:00 MHDA- 2005/09/13 09:00 CRDT- 2005/05/04 09:00 PHST- 2005/05/04 09:00 [pubmed] PHST- 2005/09/13 09:00 [medline] PHST- 2005/05/04 09:00 [entrez] AID - S0021-9258(20)56836-1 [pii] AID - 10.1074/jbc.M501988200 [doi] PST - ppublish SO - J Biol Chem. 2005 Jul 15;280(28):26435-47. doi: 10.1074/jbc.M501988200. Epub 2005 May 2.