CE15_UNKP
ID CE15_UNKP Reviewed; 432 AA.
AC A0A0K2VM55;
DT 15-MAR-2017, integrated into UniProtKB/Swiss-Prot.
DT 11-NOV-2015, sequence version 1.
DT 03-AUG-2022, entry version 15.
DE RecName: Full=Carbohydrate esterase MZ0003 {ECO:0000305|PubMed:27433797};
DE EC=3.1.1.- {ECO:0000269|PubMed:27433797};
DE Flags: Precursor;
GN Name=MZ0003 {ECO:0000312|EMBL:CDW92038.1};
OS Unknown prokaryotic organism.
OC Bacteria; environmental samples.
OX NCBI_TaxID=2725;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RA Tian C., Fan X.;
RT "Three species of the Botryosphaeriales overlap on five unrelated trees in
RT China, with a novel species.";
RL Submitted (JUN-2014) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP FUNCTION, ESTERASE ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE
RP SPECIFICITY, COFACTOR, ACTIVITY REGULATION, SUBCELLULAR LOCATION,
RP 3D-STRUCTURE MODELING, MUTAGENESIS OF 152-LYS-GLU-153; SER-268; CYS-291 AND
RP HIS-409, AND ACTIVE SITE.
RX PubMed=27433797; DOI=10.1371/journal.pone.0159345;
RA De Santi C., Willassen N.P., Williamson A.;
RT "Biochemical characterization of a family 15 carbohydrate esterase from a
RT bacterial marine arctic metagenome.";
RL PLoS ONE 11:E0159345-E0159345(2016).
CC -!- FUNCTION: Displays some glucuronoyl esterase activity in vitro, since
CC it is able to hydrolyze methyl 4-O-methyl-D-glucopyranosyluronate,
CC allyl D-glucuronate, benzyl D-glucuronate and D-glucuronic acid methyl
CC ester. However, esters of glucuronic acid are probably not its
CC biological substrate, as they are not present in the marine
CC environment. Can also hydrolyze a range of other esters, including p-
CC nitrophenyl acetate. More likely biologically-relevant substrates for
CC MZ0003 and other marine bacterial CE15s are algal cell wall
CC polysaccharides, as these would be readily available in this
CC environment and could be used as energy sources.
CC {ECO:0000269|PubMed:27433797}.
CC -!- COFACTOR:
CC Note=Does not require metal ions for activity.
CC {ECO:0000269|PubMed:27433797};
CC -!- ACTIVITY REGULATION: Is inhibited by PMSF and by NaF in vitro, which is
CC consistent with the catalytic nucleophile being a serine.
CC {ECO:0000269|PubMed:27433797}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.71 mM for p-nitrophenyl acetate (in the absence of NaCl)
CC {ECO:0000269|PubMed:27433797};
CC KM=0.97 mM for p-nitrophenyl acetate (in the presence of 1M NaCl)
CC {ECO:0000269|PubMed:27433797};
CC Note=kcat is 8.9 sec(-1) with p-nitrophenyl acetate as substrate (in
CC the absence of NaCl). kcat is 12.78 sec(-1) with p-nitrophenyl
CC acetate as substrate (in the presence of 1M NaCl).
CC {ECO:0000269|PubMed:27433797};
CC pH dependence:
CC Optimum pH is 8.0. Shows more than 60% of its maximum activity in the
CC pH range of 7.0-9.5. 20% of the activity is retained at low pH range
CC and at pH 10.5, while no catalytic activity is found at pH 5.0 or
CC 10.5. {ECO:0000269|PubMed:27433797};
CC Temperature dependence:
CC Optimum temperature is 35 degrees Celsius with p-nitrophenyl acetate
CC as substrate. Above 35 degrees Celsius, activity significantly
CC decreases, while 30% of the maximum activity is recorded at 10
CC degrees Celsius. {ECO:0000269|PubMed:27433797};
CC -!- SUBCELLULAR LOCATION: Periplasm {ECO:0000305|PubMed:27433797}.
CC -!- SIMILARITY: Belongs to the carbohydrate esterase 15 (CE15) family.
CC {ECO:0000305|PubMed:27433797}.
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DR EMBL; LM651246; CDW92038.1; -; Genomic_DNA.
DR PDB; 6EHN; X-ray; 1.90 A; A=29-427.
DR PDBsum; 6EHN; -.
DR AlphaFoldDB; A0A0K2VM55; -.
DR SMR; A0A0K2VM55; -.
DR ESTHER; unkp-CE15; Glucuronoyl_esterase.
DR GO; GO:0042597; C:periplasmic space; IEA:UniProtKB-SubCell.
DR GO; GO:0052689; F:carboxylic ester hydrolase activity; IDA:UniProtKB.
DR GO; GO:0000272; P:polysaccharide catabolic process; IEA:UniProtKB-KW.
DR Gene3D; 3.40.50.1820; -; 1.
DR InterPro; IPR029058; AB_hydrolase.
DR SUPFAM; SSF53474; SSF53474; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Carbohydrate metabolism; Hydrolase; Periplasm;
KW Polysaccharide degradation; Serine esterase; Signal.
FT SIGNAL 1..25
FT /evidence="ECO:0000255"
FT CHAIN 26..432
FT /note="Carbohydrate esterase MZ0003"
FT /id="PRO_0000439191"
FT MOTIF 266..271
FT /note="GXSYXG catalytic site motif"
FT /evidence="ECO:0000250|UniProtKB:G2QJR6"
FT ACT_SITE 268
FT /note="Nucleophile"
FT /evidence="ECO:0000305|PubMed:27433797"
FT ACT_SITE 409
FT /note="Charge relay system"
FT /evidence="ECO:0000305|PubMed:27433797"
FT BINDING 272
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:G2QJR6"
FT BINDING 359
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:G2QJR6"
FT MUTAGEN 152..153
FT /note="KE->AA: No effect on esterase activity."
FT /evidence="ECO:0000269|PubMed:27433797"
FT MUTAGEN 268
FT /note="S->A: Loss of esterase activity."
FT /evidence="ECO:0000269|PubMed:27433797"
FT MUTAGEN 291
FT /note="C->A: No effect on esterase activity."
FT /evidence="ECO:0000269|PubMed:27433797"
FT MUTAGEN 409
FT /note="H->A: Loss of esterase activity."
FT /evidence="ECO:0000269|PubMed:27433797"
FT HELIX 34..36
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 57..62
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 64..75
FT /evidence="ECO:0007829|PDB:6EHN"
FT STRAND 87..89
FT /evidence="ECO:0007829|PDB:6EHN"
FT STRAND 95..97
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 98..100
FT /evidence="ECO:0007829|PDB:6EHN"
FT STRAND 102..114
FT /evidence="ECO:0007829|PDB:6EHN"
FT STRAND 119..127
FT /evidence="ECO:0007829|PDB:6EHN"
FT STRAND 134..143
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 145..147
FT /evidence="ECO:0007829|PDB:6EHN"
FT TURN 165..168
FT /evidence="ECO:0007829|PDB:6EHN"
FT STRAND 171..173
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 176..178
FT /evidence="ECO:0007829|PDB:6EHN"
FT TURN 179..182
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 183..185
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 188..193
FT /evidence="ECO:0007829|PDB:6EHN"
FT STRAND 197..202
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 203..206
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 217..222
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 230..232
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 236..253
FT /evidence="ECO:0007829|PDB:6EHN"
FT STRAND 257..265
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 269..280
FT /evidence="ECO:0007829|PDB:6EHN"
FT STRAND 285..291
FT /evidence="ECO:0007829|PDB:6EHN"
FT TURN 294..297
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 300..302
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 308..314
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 321..326
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 330..332
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 337..343
FT /evidence="ECO:0007829|PDB:6EHN"
FT TURN 344..346
FT /evidence="ECO:0007829|PDB:6EHN"
FT STRAND 349..354
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 358..360
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 362..371
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 373..378
FT /evidence="ECO:0007829|PDB:6EHN"
FT STRAND 398..407
FT /evidence="ECO:0007829|PDB:6EHN"
FT HELIX 413..426
FT /evidence="ECO:0007829|PDB:6EHN"
SQ SEQUENCE 432 AA; 48457 MW; 57E02D3A26F662F7 CRC64;
MQRTCVLIVL IVTSTMWTPD PDVYAQPRGF NYDEAQVPKY TLPDPLVMVD GTKVTSAKQW
NDKRRDEVQQ LFEAYMYGKV PDGETELIFT DAKGERALGG AAIRKQVKIS FGEKEDAPAM
DLLIYLPADA KVRVPVFLGL NFHGNHTIHK DKEIWLTESW VRTNKKFGIT KNKANELSRG
VAAGRWQIEK AIAKGYGVAT IYCGDIDPDF NFPSNGIQAY YYKKDQTIPE KGQWGTIAAW
AFGLSCAMDY FETDTDIDHK KVAVLGHSRL GKTSLWAGAI DTRFALTISN CSGCGGAALS
RRRFGETVRR INTSFPHWFC SRFHQYNDKE DKLPIDQHML IALCAPRPVL INSATEDKWA
DPHGEFLAAQ GADAVYRMLG TGGLDAKKWP EPNKLVKSTI GYHLRPGKHD VTARDWDVYI
EFADHHMTGG AE
