RSAKS_PSESP
ID RSAKS_PSESP Reviewed; 663 AA.
AC F8KAY7;
DT 25-MAY-2022, integrated into UniProtKB/Swiss-Prot.
DT 21-SEP-2011, sequence version 1.
DT 03-AUG-2022, entry version 45.
DE RecName: Full=(R)-specific secondary-alkylsulfatase {ECO:0000305};
DE Short=(R)-specific sec-alkylsulfatase {ECO:0000305};
DE EC=3.1.6.19 {ECO:0000269|PubMed:21770430, ECO:0000269|PubMed:23061549, ECO:0000269|Ref.2};
DE AltName: Full=Inverting sec-alkylsulfatase Pisa1 {ECO:0000303|Ref.2};
DE AltName: Full=Inverting secondary alkylsulfatase 1 {ECO:0000303|PubMed:23061549};
DE AltName: Full=Type III (R)-specific secondary-alkylsulfatase {ECO:0000305};
DE Flags: Precursor;
GN Name=pisa1 {ECO:0000303|PubMed:21770430};
OS Pseudomonas sp.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas.
OX NCBI_TaxID=306;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, AND CATALYTIC ACTIVITY.
RC STRAIN=DSM 6611;
RX PubMed=21770430; DOI=10.1021/ol201635y;
RA Schober M., Gadler P., Knaus T., Kayer H., Birner-Gruenberger R.,
RA Guelly C., Macheroux P., Wagner U., Faber K.;
RT "A stereoselective inverting sec-alkylsulfatase for the deracemization of
RT sec-alcohols.";
RL Org. Lett. 13:4296-4299(2011).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=DSM 6611;
RX DOI=10.1002/adsc.201100864;
RA Schober M., Knaus T., Toesch M., Macheroux P., Wagner U., Faber K.;
RT "The substrate spectrum of the inverting sec-alkylsulfatase Pisa1.";
RL Adv. Synth. Catal. 354:1737-1742(2012).
RN [3] {ECO:0007744|PDB:2YHE, ECO:0007744|PDB:4AV7, ECO:0007744|PDB:4AXH}
RP X-RAY CRYSTALLOGRAPHY (2.70 ANGSTROMS) OF 1-660 IN COMPLEX WITH ZINC AND
RP SULFATE, FUNCTION, CATALYTIC ACTIVITY, REACTION MECHANISM,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, AND MUTAGENESIS OF SER-233;
RP PHE-250; HIS-317 AND TYR-417.
RC STRAIN=DSM 6611;
RX PubMed=23061549; DOI=10.1111/febs.12027;
RA Knaus T., Schober M., Kepplinger B., Faccinelli M., Pitzer J., Faber K.,
RA Macheroux P., Wagner U.;
RT "Structure and mechanism of an inverting alkylsulfatase from Pseudomonas
RT sp. DSM6611 specific for secondary alkyl sulfates.";
RL FEBS J. 279:4374-4384(2012).
CC -!- FUNCTION: Alkylsulfatase that catalyzes the enantioselective hydrolysis
CC of secondary-alkylsulfates with strict inversion of configuration,
CC leading to the formation of homochiral (S)-configurated alcohols and
CC nonreacted sulfate esters (PubMed:21770430, Ref.2, PubMed:23061549).
CC The substrate spectrum includes a range of linear, branched or cyclic
CC sec-alkylsulfates (PubMed:21770430). Can use sec-alkylsulfate esters
CC bearing aromatic, olefinic and acetylenic moieties (Ref.2). Acts by
CC cleaving the C-O bond, resulting in inversion at the carbon
CC (PubMed:23061549). {ECO:0000269|PubMed:21770430,
CC ECO:0000269|PubMed:23061549, ECO:0000269|Ref.2}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=an (R)-secondary-alkyl sulfate + H2O = an (S)-secondary-
CC alcohol + sulfate.; EC=3.1.6.19;
CC Evidence={ECO:0000269|PubMed:21770430, ECO:0000269|PubMed:23061549,
CC ECO:0000269|Ref.2};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=151 uM for (R)-2-octyl sulfate {ECO:0000269|PubMed:23061549};
CC KM=651 uM for 1-octyl sulfate {ECO:0000269|PubMed:23061549};
CC Note=kcat is 262 min(-1) with (R)-2-octyl sulfate as substrate. kcat
CC is 52 min(-1) with 1-octyl sulfate as substrate.
CC {ECO:0000269|PubMed:23061549};
CC Temperature dependence:
CC Is very stable at 20-30 degrees Celsius. {ECO:0000269|Ref.2};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:23061549}.
CC -!- MISCELLANEOUS: Is a very useful inverting alkylsulfatase for the
CC deracemisation of rac-sec-alcohols via enzymatic hydrolysis of their
CC corresponding sulfate esters, which furnishes homochiral products.
CC {ECO:0000269|Ref.2}.
CC -!- SIMILARITY: Belongs to the metallo-beta-lactamase superfamily. Type III
CC sulfatase family. {ECO:0000305}.
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DR EMBL; FR850678; CCA63329.1; -; Genomic_DNA.
DR PDB; 2YHE; X-ray; 2.70 A; A/B/C/D/E/F=1-660.
DR PDB; 4AV7; X-ray; 3.00 A; A/B/C/D/E/F=1-660.
DR PDB; 4AXH; X-ray; 2.70 A; A/B=1-660.
DR PDBsum; 2YHE; -.
DR PDBsum; 4AV7; -.
DR PDBsum; 4AXH; -.
DR SMR; F8KAY7; -.
DR MINT; F8KAY7; -.
DR BRENDA; 3.1.6.19; 17955.
DR GO; GO:0018741; F:alkyl sulfatase activity; IEA:InterPro.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0046983; F:protein dimerization activity; IEA:InterPro.
DR GO; GO:0018909; P:dodecyl sulfate metabolic process; IEA:InterPro.
DR CDD; cd07710; arylsulfatase_Sdsa1-like_MBL-fold; 1.
DR Gene3D; 1.25.40.880; -; 1.
DR Gene3D; 3.30.1050.10; -; 1.
DR InterPro; IPR038536; Alkyl/aryl-sulf_dimr_sf.
DR InterPro; IPR029229; Alkyl_sulf_C.
DR InterPro; IPR029228; Alkyl_sulf_dimr.
DR InterPro; IPR044097; Bds1/SdsA1_MBL-fold.
DR InterPro; IPR001279; Metallo-B-lactamas.
DR InterPro; IPR036866; RibonucZ/Hydroxyglut_hydro.
DR InterPro; IPR036527; SCP2_sterol-bd_dom_sf.
DR Pfam; PF14864; Alkyl_sulf_C; 1.
DR Pfam; PF14863; Alkyl_sulf_dimr; 1.
DR Pfam; PF00753; Lactamase_B; 1.
DR SMART; SM00849; Lactamase_B; 1.
DR SUPFAM; SSF55718; SSF55718; 1.
DR SUPFAM; SSF56281; SSF56281; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Hydrolase; Metal-binding; Signal; Zinc.
FT SIGNAL 1..28
FT /evidence="ECO:0000255"
FT CHAIN 29..663
FT /note="(R)-specific secondary-alkylsulfatase"
FT /id="PRO_5005678730"
FT BINDING 179
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:23061549,
FT ECO:0007744|PDB:2YHE, ECO:0007744|PDB:4AV7,
FT ECO:0007744|PDB:4AXH"
FT BINDING 181
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:23061549,
FT ECO:0007744|PDB:2YHE, ECO:0007744|PDB:4AV7,
FT ECO:0007744|PDB:4AXH"
FT BINDING 183
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:23061549,
FT ECO:0007744|PDB:2YHE, ECO:0007744|PDB:4AV7,
FT ECO:0007744|PDB:4AXH"
FT BINDING 184
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:23061549,
FT ECO:0007744|PDB:2YHE, ECO:0007744|PDB:4AV7,
FT ECO:0007744|PDB:4AXH"
FT BINDING 232
FT /ligand="sulfate"
FT /ligand_id="ChEBI:CHEBI:16189"
FT /evidence="ECO:0000269|PubMed:23061549,
FT ECO:0007744|PDB:2YHE, ECO:0007744|PDB:4AV7,
FT ECO:0007744|PDB:4AXH"
FT BINDING 291
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:23061549,
FT ECO:0007744|PDB:2YHE, ECO:0007744|PDB:4AV7,
FT ECO:0007744|PDB:4AXH"
FT BINDING 310
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:23061549,
FT ECO:0007744|PDB:2YHE, ECO:0007744|PDB:4AV7,
FT ECO:0007744|PDB:4AXH"
FT BINDING 310
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:23061549,
FT ECO:0007744|PDB:2YHE, ECO:0007744|PDB:4AV7,
FT ECO:0007744|PDB:4AXH"
FT BINDING 318..323
FT /ligand="sulfate"
FT /ligand_id="ChEBI:CHEBI:16189"
FT /evidence="ECO:0000269|PubMed:23061549,
FT ECO:0007744|PDB:2YHE, ECO:0007744|PDB:4AV7,
FT ECO:0007744|PDB:4AXH"
FT BINDING 328
FT /ligand="sulfate"
FT /ligand_id="ChEBI:CHEBI:16189"
FT /evidence="ECO:0000269|PubMed:23061549,
FT ECO:0007744|PDB:2YHE, ECO:0007744|PDB:4AV7,
FT ECO:0007744|PDB:4AXH"
FT BINDING 355
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:23061549,
FT ECO:0007744|PDB:2YHE, ECO:0007744|PDB:4AV7,
FT ECO:0007744|PDB:4AXH"
FT BINDING 417
FT /ligand="sulfate"
FT /ligand_id="ChEBI:CHEBI:16189"
FT /evidence="ECO:0000269|PubMed:23061549,
FT ECO:0007744|PDB:2YHE, ECO:0007744|PDB:4AV7,
FT ECO:0007744|PDB:4AXH"
FT MUTAGEN 233
FT /note="S->Y: 2.5-fold decrease in kcat with (R)-2-octyl
FT sulfate as substrate. Cannot use (S)-2-octyl sulfate. Shows
FT activity towards (S)-2-octyl sulfate; when associated with
FT G-250."
FT /evidence="ECO:0000269|PubMed:23061549"
FT MUTAGEN 250
FT /note="F->G: 190-fold decrease in kcat with (R)-2-octyl
FT sulfate as substrate. 28-fold increase in Km for (R)-2-
FT octyl sulfate. Cannot use (S)-2-octyl sulfate. Shows
FT activity towards (S)-2-octyl sulfate; when associated with
FT Y-233."
FT /evidence="ECO:0000269|PubMed:23061549"
FT MUTAGEN 317
FT /note="H->A: 1300-fold decrease in kcat with (R)-2-octyl
FT sulfate as substrate."
FT /evidence="ECO:0000269|PubMed:23061549"
FT MUTAGEN 417
FT /note="Y->D: Loss of activity."
FT /evidence="ECO:0000269|PubMed:23061549"
FT MUTAGEN 417
FT /note="Y->F: 114-fold decrease in kcat with (R)-2-octyl
FT sulfate as substrate. 65-fold increase in Km for (R)-2-
FT octyl sulfate."
FT /evidence="ECO:0000269|PubMed:23061549"
FT MUTAGEN 417
FT /note="Y->H: 238-fold decrease in kcat with (R)-2-octyl
FT sulfate as substrate. 13-fold increase in Km for (R)-2-
FT octyl sulfate."
FT /evidence="ECO:0000269|PubMed:23061549"
SQ SEQUENCE 663 AA; 73142 MW; 682966688172CBA1 CRC64;
MSRFIRASQR RTLLATLIAA TLAQPLLAAE SLDSKPASAI TAAKNAEVLK NLPFADREEF
EAAKRGLIAP FSGQIKNAEG QVVWDMGAYQ FLNDKDAADT VNPSLWHQAQ LNNIAGLFEV
MPKLYQVRGL DPANMTIIEG DSGLVLIDTL TTAETARAAL DLYFQHRPKK PIVAVVYSHS
HIDHFGGARG IIDEADVKAG KVKVFAPSGF MEHAVSENIL AGTAMARRGQ YQSGVMVPRG
AQAQVDSGLF KTTATNATNT LVAPNVLIEK PYERHTVDGV ELEFQLTLGS EAPSDMNIYL
PQFKVLNTAD NAPPAMHNLL TPRGAEVRDA KAWAGYIDAS LEKYGDRTDV LIQQHNWPVW
GGDKVRTYLA DQRDMYAFLN NRALNLMNKG LTLHEIAAEV SKLPGELDRK WYLRSYYGAL
STNLRAVYQR YLGFYDGNPA NLDPFPPVEA GKRYVEAMGG ADAVLKQMRA AIDKGDYRWA
VQLGNHLVFA DPANKDARAL QADAMEQLGY QTENALWRNM YMTGAMELRH GVPTYDSRGK
SEMGRALTPD MFFDLLAIRL DTDKAVGHDM TLNWVFEDLK QDIALTLRNG VLTQRVGSLN
PKADVTVKLT KPTLDQIAAR KLDLPTAIKQ GTVKLDGDGK KLGEFFGLLD SFSPKFNIVE
PLE
