ID   PHAM_XANAG              Reviewed;         453 AA.
AC   P0C2Y0;
DT   15-MAY-2007, integrated into UniProtKB/Swiss-Prot.
DT   15-MAY-2007, sequence version 1.
DT   25-MAY-2022, entry version 27.
DE   RecName: Full=o-phthalyl amidase;
DE            EC=3.5.1.79;
OS   Xanthobacter agilis.
OC   Bacteria; Proteobacteria; Alphaproteobacteria; Hyphomicrobiales;
OC   Xanthobacteraceae; Xanthobacter.
OX   NCBI_TaxID=47492;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, AND BIOPHYSICOCHEMICAL
RP   PROPERTIES.
RA   Briggs B.S., Zmijewski M.J. Jr.;
RT   "Enzyme from microbial source: phthalyl amidase.";
RL   Patent number US5445959, 29-AUG-1995.
RN   [2]
RP   PROTEIN SEQUENCE OF 143-168; 210-236; 267-281; 323-341 AND 435-447,
RP   FUNCTION, SUBSTRATE SPECIFICITY, SUBUNIT, BLOCKAGE OF N-TERMINUS, ACTIVITY
RP   REGULATION, AND BIOPHYSICOCHEMICAL PROPERTIES.
RA   Briggs B.S., Kreuzman A.J., Whitesitt C., Yeh W.-K., Zmijewski M.J. Jr.;
RT   "Discovery, purification, and properties of o-phthalyl amidase from
RT   Xanthobacter agilis.";
RL   J. Mol. Catal., B Enzym. 2:53-69(1996).
CC   -!- FUNCTION: Catalyzes the removal of the phthalyl group from phthalyl
CC       amides generating phthalate and an amine. The enzyme has a broad
CC       substrate specificity and hydrolyzes phthalylated amino acids,
CC       peptides, beta-lactams, aromatic and aliphatic amines; substitutions
CC       allowed on the phthalyl group include 6-F, 6-NH(2), 3-OH, and a
CC       nitrogen in the aromatic ring ortho to the carboxy group attached to
CC       the amine. {ECO:0000269|Ref.1, ECO:0000269|Ref.2}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=a phtalamide + H2O = a primary amine + phthalate;
CC         Xref=Rhea:RHEA:18297, ChEBI:CHEBI:15377, ChEBI:CHEBI:17563,
CC         ChEBI:CHEBI:65295, ChEBI:CHEBI:65296; EC=3.5.1.79;
CC   -!- ACTIVITY REGULATION: Inhibited by iodoacetate, p-hydroxymercuric
CC       benzoate and copper ions. {ECO:0000269|Ref.2}.
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=0.9 mM for o-phthalyl carbacephem {ECO:0000269|Ref.1,
CC         ECO:0000269|Ref.2};
CC         KM=0.05 mM for 4-(2'-carboxy-N-benzoyl)amino-2-carboxy-nitrobenzene
CC         {ECO:0000269|Ref.1, ECO:0000269|Ref.2};
CC         Vmax=7.6 umol/min/mg enzyme with o-phthalyl carbacephem as substrate
CC         {ECO:0000269|Ref.1, ECO:0000269|Ref.2};
CC         Vmax=5.95 umol/min/mg enzyme with 4-(2'-carboxy-N-benzoyl)amino-2-
CC         carboxy-nitrobenzene as substrate {ECO:0000269|Ref.1,
CC         ECO:0000269|Ref.2};
CC       pH dependence:
CC         Optimum pH is 7.8-8.4. {ECO:0000269|Ref.1, ECO:0000269|Ref.2};
CC       Temperature dependence:
CC         Optimum temperature is 28-34 degrees Celsius. {ECO:0000269|Ref.1,
CC         ECO:0000269|Ref.2};
CC   -!- SUBUNIT: Monomer. {ECO:0000269|Ref.2}.
CC   -!- PTM: The N-terminus is blocked.
CC   -!- BIOTECHNOLOGY: The broad substrate acceptance, high catalytic activity,
CC       and stability at high salt or substrate concentration of the enzyme
CC       indicates that it can serve as a gentle method for deprotecting
CC       phthalimido and o-phthalyl protected amides in new chemo-enzymatic
CC       synthetic routes.
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DR   EMBL; I14148; -; NOT_ANNOTATED_CDS; Genomic_DNA.
DR   AlphaFoldDB; P0C2Y0; -.
DR   SMR; P0C2Y0; -.
DR   GO; GO:0047418; F:phthalyl amidase activity; IEA:UniProtKB-EC.
DR   Gene3D; 3.40.50.1820; -; 1.
DR   InterPro; IPR029058; AB_hydrolase.
DR   InterPro; IPR045556; DUF6351.
DR   Pfam; PF19878; DUF6351; 1.
DR   SUPFAM; SSF53474; SSF53474; 1.
PE   1: Evidence at protein level;
KW   Direct protein sequencing; Hydrolase.
FT   CHAIN           1..453
FT                   /note="o-phthalyl amidase"
FT                   /id="PRO_0000287884"
SQ   SEQUENCE   453 AA;  49856 MW;  E48F11725D2DFF42 CRC64;
     MQAPSVHQHV AFTEEIGDLP DGSSYMIRVP ENWNGVLIRD LDLVSGTSNS NAARYETMLK
     EGFAVAGTAR HPLRQWQYDP AHEIENLNHV LDTFEENYGS PERVIQYGCS GGAHVSLAVA
     EDFSDRVDGS VALAAHTPVW IMNSFLDGWF SLQSLIGEYY VEAGHGPLSD LAITKLPNDG
     SSNSSGHGME GDLPAAWRNA FTAANATPEG RARMALAFAL GQWSPWLADN TPQPDLDDPE
     AIADSVYESA MRLAGSPGGE ARIMFENAAR GQQLSWNDDI DYADFWENSN PAMKSAVQEL
     YDTAGLDLQS DIETVNSQPR IEASQYALDY WNTPGRNVIG DPEVPVLRLH MIGDYQIPYS
     LVQGYSDLIS ENNNDDLYRT AFVQSTGHCN FTAAESSAAI EVMMQRLDTG EWPSTEPDDL
     NAIAEASNTG TEARFMALDG WEIPEYNRTW KPE