{"@context":{"@vocab":"https://cir.nii.ac.jp/schema/1.0/","rdfs":"http://www.w3.org/2000/01/rdf-schema#","dc":"http://purl.org/dc/elements/1.1/","dcterms":"http://purl.org/dc/terms/","foaf":"http://xmlns.com/foaf/0.1/","prism":"http://prismstandard.org/namespaces/basic/2.0/","cinii":"http://ci.nii.ac.jp/ns/1.0/","datacite":"https://schema.datacite.org/meta/kernel-4/","ndl":"http://ndl.go.jp/dcndl/terms/","jpcoar":"https://github.com/JPCOAR/schema/blob/master/2.0/"},"@id":"https://cir.nii.ac.jp/crid/1361699994385806080.json","@type":"Article","productIdentifier":[{"identifier":{"@type":"DOI","@value":"10.1128/jb.121.1.272-285.1975"}},{"identifier":{"@type":"URI","@value":"https://journals.asm.org/doi/pdf/10.1128/jb.121.1.272-285.1975"}}],"dc:title":[{"@value":"Metabolic function and properties of 4-hydroxyphenylacetic acid 1-hydroxylase from Pseudomonas acidovorans"}],"description":[{"type":"abstract","notation":[{"@value":"<jats:p>The enzyme 4-hydroxyphenylacetate, NAD(P)H:oxygen oxidoreductase (1-hydroxylating) (EC 1.14.13 ...; 4-hydroxyphenylacetate 1-monooxygenase; referred to here as 4-HPA 1-hydroxylase) was induced in Pseudomonas acidovorans when 4-hydroxyphenylacetate (4-PHA) was utilized as carbon source for growth; homogentisate and maleylacetoacetate were intermediates in the degradation of 4-HPA. A preparation of the hydroxylase that was free from homogentisate dioxygenase and could be stored at 4 C in the presence of dithioerythritol with little loss of activity was obtained by ultracentrifuging cell extracts; but when purified 18-fold by affinity chromatography the enzyme became unstable. Flavin adenine dinucleotide and Mg2+ ions were required for full activity. 4-HPA 1-hydrocylase was inhibited by KCl, which was uncompetitive with 4-HPA. Values of Ki determined for inhibitors competitive with 4-HPA were 17 muM dl-4-hydroxymandelic acid, 43 muM 3,4-dihydroxyphenylacetic acid, 87 muM 4-hydroxy-3-methylphenylacetic acid, and 440 muM 4-hydroxyphenylpropionic acid. Apparent Km values for substrates of 4-HPA 1-hydroxylase were 31 muM 4-HPA, 67 muM oxygen, 95 muM reduced nicotinamide adenine dinucleotide (NADH); AND 250 muM reduced nicotinamide adenine dinucleotide phosphate (NADPH). The same maximum velocity was given by NADH and NADPH. A chemical synthesis is described for 2-deutero-4-hydroxyphenylacetic acid. This compound was enzymatically hydroxylated with retention of half the deuterium in the homogentisic acid formed. Activity as substrate or inhibitor of 4-HPA 1-hydroxylase was shown only by those analogues of 4-HPA that possessed a hydroxyl group substituent at C-4 of the benze nucleus. A mechanism is suggested that accounts for this structural requirement and also for the observation that when 4-hydroxyphenoxyacetic acid was attacked by the enzyme, hydroquinone was formed by release of the side chain, probably as glycolic acid. Only one enantiometer of racemic 4-hydroxyhydratropic acid was attacked by 4-HPA 1-hydroxylase; the product, alpha-methylhomogentisic acid (2-(2,5-dihydroxyphenyl)-propionic acid), exhibited optical activity. This observation suggests that, during its shift from C-1 to C-2 of the nucleus, the side chain of the substrate remains bound to a site on the enzyme while a conformational change of the protein permits the necessary movement of the benzene ring.</jats:p>"}]}],"creator":[{"@id":"https://cir.nii.ac.jp/crid/1381699994385806082","@type":"Researcher","foaf:name":[{"@value":"W A Hareland"}]},{"@id":"https://cir.nii.ac.jp/crid/1381699994385806080","@type":"Researcher","foaf:name":[{"@value":"R L Crawford"}]},{"@id":"https://cir.nii.ac.jp/crid/1381699994385806083","@type":"Researcher","foaf:name":[{"@value":"P J Chapman"}]},{"@id":"https://cir.nii.ac.jp/crid/1381699994385806081","@type":"Researcher","foaf:name":[{"@value":"S Dagley"}]}],"publication":{"publicationIdentifier":[{"@type":"PISSN","@value":"00219193"},{"@type":"EISSN","@value":"10985530"}],"prism:publicationName":[{"@value":"Journal of Bacteriology"}],"dc:publisher":[{"@value":"American Society for Microbiology"}],"prism:publicationDate":"1975-01","prism:volume":"121","prism:number":"1","prism:startingPage":"272","prism:endingPage":"285"},"reviewed":"false","dc:rights":["https://journals.asm.org/non-commercial-tdm-license"],"url":[{"@id":"https://journals.asm.org/doi/pdf/10.1128/jb.121.1.272-285.1975"}],"createdAt":"2020-01-03","modifiedAt":"2021-07-29","relatedProduct":[{"@id":"https://cir.nii.ac.jp/crid/1360284918863362560","@type":"Article","relationType":["isReferencedBy"],"jpcoar:relatedTitle":[{"@value":"Purification and characterization of o-hydroxyphenylacetate 5-hydroxylase, m-hydroxyphenylacetate 6-hydroxylase and p-hydroxyphenylacetate 1-hydroxylase from Rhodococcus erythropolis"}]}],"dataSourceIdentifier":[{"@type":"CROSSREF","@value":"10.1128/jb.121.1.272-285.1975"},{"@type":"CROSSREF","@value":"10.1016/0922-338x(96)87590-8_references_DOI_O6YGeanRcr6HGNHzztOoO2Z0T8P"}]}