{"entryType":"UniProtKB reviewed (Swiss-Prot)","primaryAccession":"P0C2Y0","uniProtkbId":"PHAM_XANAG","entryAudit":{"firstPublicDate":"2007-05-15","lastAnnotationUpdateDate":"2025-04-09","lastSequenceUpdateDate":"2007-05-15","entryVersion":32,"sequenceVersion":1},"annotationScore":4.0,"organism":{"scientificName":"Xanthobacter agilis","taxonId":47492,"lineage":["Bacteria","Pseudomonadati","Pseudomonadota","Alphaproteobacteria","Hyphomicrobiales","Xanthobacteraceae","Xanthobacter"]},"proteinExistence":"1: Evidence at protein level","proteinDescription":{"recommendedName":{"fullName":{"value":"o-phthalyl amidase"},"ecNumbers":[{"value":"3.5.1.79"}]}},"comments":[{"texts":[{"evidences":[{"evidenceCode":"ECO:0000269","source":"Reference","id":"Ref.1"},{"evidenceCode":"ECO:0000269","source":"Reference","id":"Ref.2"}],"value":"Catalyzes the removal of the phthalyl group from phthalyl amides generating phthalate and an amine. The enzyme has a broad substrate specificity and hydrolyzes phthalylated amino acids, peptides, beta-lactams, aromatic and aliphatic amines; substitutions allowed on the phthalyl group include 6-F, 6-NH(2), 3-OH, and a nitrogen in the aromatic ring ortho to the carboxy group attached to the amine"}],"commentType":"FUNCTION"},{"commentType":"CATALYTIC ACTIVITY","reaction":{"name":"a phtalamide + H2O = phthalate + a primary amine","reactionCrossReferences":[{"database":"Rhea","id":"RHEA:18297"},{"database":"ChEBI","id":"CHEBI:15377"},{"database":"ChEBI","id":"CHEBI:17563"},{"database":"ChEBI","id":"CHEBI:65295"},{"database":"ChEBI","id":"CHEBI:65296"}],"ecNumber":"3.5.1.79"}},{"texts":[{"evidences":[{"evidenceCode":"ECO:0000269","source":"Reference","id":"Ref.2"}],"value":"Inhibited by iodoacetate, p-hydroxymercuric benzoate and copper ions"}],"commentType":"ACTIVITY REGULATION"},{"commentType":"BIOPHYSICOCHEMICAL PROPERTIES","kineticParameters":{"maximumVelocities":[{"velocity":7.6,"unit":"umol/min/mg","enzyme":"enzyme with o-phthalyl carbacephem as substrate","evidences":[{"evidenceCode":"ECO:0000269","source":"Reference","id":"Ref.1"},{"evidenceCode":"ECO:0000269","source":"Reference","id":"Ref.2"}]},{"velocity":5.95,"unit":"umol/min/mg","enzyme":"enzyme with 4-(2'-carboxy-N-benzoyl)amino-2-carboxy-nitrobenzene as substrate","evidences":[{"evidenceCode":"ECO:0000269","source":"Reference","id":"Ref.1"},{"evidenceCode":"ECO:0000269","source":"Reference","id":"Ref.2"}]}],"michaelisConstants":[{"constant":0.9,"unit":"mM","substrate":"o-phthalyl carbacephem","evidences":[{"evidenceCode":"ECO:0000269","source":"Reference","id":"Ref.1"},{"evidenceCode":"ECO:0000269","source":"Reference","id":"Ref.2"}]},{"constant":0.05,"unit":"mM","substrate":"4-(2'-carboxy-N-benzoyl)amino-2-carboxy-nitrobenzene","evidences":[{"evidenceCode":"ECO:0000269","source":"Reference","id":"Ref.1"},{"evidenceCode":"ECO:0000269","source":"Reference","id":"Ref.2"}]}]},"phDependence":{"texts":[{"evidences":[{"evidenceCode":"ECO:0000269","source":"Reference","id":"Ref.1"},{"evidenceCode":"ECO:0000269","source":"Reference","id":"Ref.2"}],"value":"Optimum pH is 7.8-8.4."}]},"temperatureDependence":{"texts":[{"evidences":[{"evidenceCode":"ECO:0000269","source":"Reference","id":"Ref.1"},{"evidenceCode":"ECO:0000269","source":"Reference","id":"Ref.2"}],"value":"Optimum temperature is 28-34 degrees Celsius."}]}},{"texts":[{"evidences":[{"evidenceCode":"ECO:0000269","source":"Reference","id":"Ref.2"}],"value":"Monomer"}],"commentType":"SUBUNIT"},{"texts":[{"value":"The N-terminus is blocked"}],"commentType":"PTM"},{"texts":[{"value":"The broad substrate acceptance, high catalytic activity, and stability at high salt or substrate concentration of the enzyme indicates that it can serve as a gentle method for deprotecting phthalimido and o-phthalyl protected amides in new chemo-enzymatic synthetic routes"}],"commentType":"BIOTECHNOLOGY"}],"features":[{"type":"Chain","location":{"start":{"value":1,"modifier":"EXACT"},"end":{"value":453,"modifier":"EXACT"}},"description":"o-phthalyl amidase","featureId":"PRO_0000287884"}],"keywords":[{"id":"KW-0903","category":"Technical term","name":"Direct protein sequencing"},{"id":"KW-0378","category":"Molecular function","name":"Hydrolase"}],"references":[{"referenceNumber":1,"citation":{"id":"CI-662G383F4H7KU","citationType":"patent","authors":["Briggs B.S.","Zmijewski M.J. Jr."],"title":"Enzyme from microbial source: phthalyl amidase.","publicationDate":"29-AUG-1995","patentNumber":"US5445959"},"referencePositions":["NUCLEOTIDE SEQUENCE [GENOMIC DNA]","FUNCTION","BIOPHYSICOCHEMICAL PROPERTIES"]},{"referenceNumber":2,"citation":{"id":"CI-DH9S5V85HSHS2","citationType":"journal article","authors":["Briggs B.S.","Kreuzman A.J.","Whitesitt C.","Yeh W.-K.","Zmijewski M.J. Jr."],"title":"Discovery, purification, and properties of o-phthalyl amidase from Xanthobacter agilis.","publicationDate":"1996","journal":"J. Mol. Catal., B Enzym.","firstPage":"53","lastPage":"69","volume":"2"},"referencePositions":["PROTEIN SEQUENCE OF 143-168; 210-236; 267-281; 323-341 AND 435-447","FUNCTION","SUBSTRATE SPECIFICITY","SUBUNIT","BLOCKAGE OF N-TERMINUS","ACTIVITY REGULATION","BIOPHYSICOCHEMICAL PROPERTIES"]}],"uniProtKBCrossReferences":[{"database":"EMBL","id":"I14148","properties":[{"key":"ProteinId","value":"-"},{"key":"Status","value":"NOT_ANNOTATED_CDS"},{"key":"MoleculeType","value":"Genomic_DNA"}]},{"database":"AlphaFoldDB","id":"P0C2Y0","properties":[{"key":"Description","value":"-"}]},{"database":"SMR","id":"P0C2Y0","properties":[{"key":"Description","value":"-"}]},{"database":"ESTHER","id":"xanag-pham","properties":[{"key":"FamilyName","value":"6_AlphaBeta_hydrolase"}]},{"database":"GO","id":"GO:0047418","properties":[{"key":"GoTerm","value":"F:phthalyl amidase activity"},{"key":"GoEvidenceType","value":"IEA:UniProtKB-EC"}]},{"database":"Gene3D","id":"3.40.50.1820","properties":[{"key":"EntryName","value":"alpha/beta hydrolase"},{"key":"MatchStatus","value":"1"}]},{"database":"InterPro","id":"IPR029058","properties":[{"key":"EntryName","value":"AB_hydrolase_fold"}]},{"database":"InterPro","id":"IPR045556","properties":[{"key":"EntryName","value":"DUF6351"}]},{"database":"Pfam","id":"PF19878","properties":[{"key":"EntryName","value":"DUF6351"},{"key":"MatchStatus","value":"1"}]},{"database":"SUPFAM","id":"SSF53474","properties":[{"key":"EntryName","value":"alpha/beta-Hydrolases"},{"key":"MatchStatus","value":"1"}]}],"sequence":{"value":"MQAPSVHQHVAFTEEIGDLPDGSSYMIRVPENWNGVLIRDLDLVSGTSNSNAARYETMLKEGFAVAGTARHPLRQWQYDPAHEIENLNHVLDTFEENYGSPERVIQYGCSGGAHVSLAVAEDFSDRVDGSVALAAHTPVWIMNSFLDGWFSLQSLIGEYYVEAGHGPLSDLAITKLPNDGSSNSSGHGMEGDLPAAWRNAFTAANATPEGRARMALAFALGQWSPWLADNTPQPDLDDPEAIADSVYESAMRLAGSPGGEARIMFENAARGQQLSWNDDIDYADFWENSNPAMKSAVQELYDTAGLDLQSDIETVNSQPRIEASQYALDYWNTPGRNVIGDPEVPVLRLHMIGDYQIPYSLVQGYSDLISENNNDDLYRTAFVQSTGHCNFTAAESSAAIEVMMQRLDTGEWPSTEPDDLNAIAEASNTGTEARFMALDGWEIPEYNRTWKPE","length":453,"molWeight":49856,"crc64":"E48F11725D2DFF42","md5":"FD5696EB172A288ED0A79C7875C559B0"},"extraAttributes":{"countByCommentType":{"FUNCTION":1,"CATALYTIC ACTIVITY":1,"ACTIVITY REGULATION":1,"BIOPHYSICOCHEMICAL PROPERTIES":1,"SUBUNIT":1,"PTM":1,"BIOTECHNOLOGY":1},"countByFeatureType":{"Chain":1},"uniParcId":"UPI0000142D22"}}