CEEP_RUMAL
ID CEEP_RUMAL Reviewed; 389 AA.
AC P0DKY4; A8CF79;
DT 06-MAR-2013, integrated into UniProtKB/Swiss-Prot.
DT 06-MAR-2013, sequence version 1.
DT 03-AUG-2022, entry version 39.
DE RecName: Full=Cellobiose 2-epimerase {ECO:0000255|HAMAP-Rule:MF_00929};
DE Short=CE {ECO:0000255|HAMAP-Rule:MF_00929};
DE EC=5.1.3.11 {ECO:0000255|HAMAP-Rule:MF_00929};
GN Name=ce-ne1;
OS Ruminococcus albus.
OC Bacteria; Firmicutes; Clostridia; Eubacteriales; Oscillospiraceae;
OC Ruminococcus.
OX NCBI_TaxID=1264;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-17, FUNCTION,
RP CATALYTIC ACTIVITY, AND SUBCELLULAR LOCATION.
RC STRAIN=NE1;
RX PubMed=17612504; DOI=10.1016/j.bbrc.2007.06.091;
RA Ito S., Hamada S., Yamaguchi K., Umene S., Ito H., Matsui H., Ozawa T.,
RA Taguchi H., Watanabe J., Wasaki J., Ito S.;
RT "Cloning and sequencing of the cellobiose 2-epimerase gene from an
RT obligatory anaerobe, Ruminococcus albus.";
RL Biochem. Biophys. Res. Commun. 360:640-645(2007).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-17, FUNCTION,
RP CATALYTIC ACTIVITY, ACTIVITY REGULATION, AND BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=NE1;
RX PubMed=18392616; DOI=10.1007/s00253-008-1449-7;
RA Ito S., Taguchi H., Hamada S., Kawauchi S., Ito H., Senoura T.,
RA Watanabe J., Nishimukai M., Ito S., Matsui H.;
RT "Enzymatic properties of cellobiose 2-epimerase from Ruminococcus albus and
RT the synthesis of rare oligosaccharides by the enzyme.";
RL Appl. Microbiol. Biotechnol. 79:433-441(2008).
RN [3]
RP MUTAGENESIS OF ARG-52; PHE-114; HIS-243; GLU-246; TRP-249; TRP-303;
RP TRP-304; GLU-308 AND HIS-374.
RC STRAIN=NE1;
RX PubMed=19330485; DOI=10.1007/s10529-009-9979-3;
RA Ito S., Hamada S., Ito H., Matsui H., Ozawa T., Taguchi H., Ito S.;
RT "Site-directed mutagenesis of possible catalytic residues of cellobiose 2-
RT epimerase from Ruminococcus albus.";
RL Biotechnol. Lett. 31:1065-1071(2009).
CC -!- FUNCTION: Catalyzes the reversible epimerization of cellobiose to 4-O-
CC beta-D-glucopyranosyl-D-mannose (Glc-Man). Can also epimerize
CC cellotriose to Glc-Glc-Man, cellotetraose to Glc-Glc-Glc-Man, and
CC lactose to epilactose. {ECO:0000255|HAMAP-Rule:MF_00929,
CC ECO:0000269|PubMed:17612504, ECO:0000269|PubMed:18392616}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=D-cellobiose = beta-D-glucosyl-(1->4)-D-mannopyranose;
CC Xref=Rhea:RHEA:23384, ChEBI:CHEBI:17057, ChEBI:CHEBI:47931;
CC EC=5.1.3.11; Evidence={ECO:0000255|HAMAP-Rule:MF_00929,
CC ECO:0000269|PubMed:17612504, ECO:0000269|PubMed:18392616};
CC -!- ACTIVITY REGULATION: Enhanced by Mg(2+) and Ca(2+) ions,
CC ethylenediaminetetraacetic acid, ethylene glycol tetraacetic acid and
CC citrate. Inhibited by Al(3+), Fe(3+), Co(2+), Cu(2+), Zn(2+), Pb(2+)
CC and Ag(+) ions, iodoacetate, 4-chloromercuribenzoate and N-
CC bromosuccinimide. {ECO:0000269|PubMed:18392616}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=13.8 mM for cellobiose {ECO:0000269|PubMed:18392616};
CC KM=33 mM for lactose {ECO:0000269|PubMed:18392616};
CC Vmax=88.8 umol/min/mg enzyme with cellobiose as substrate
CC {ECO:0000269|PubMed:18392616};
CC Vmax=72.5 umol/min/mg enzyme with lactose as substrate
CC {ECO:0000269|PubMed:18392616};
CC Note=kcat is 63.8 sec(-1) for cellobiose. kcat is 52.1 sec(-1) for
CC lactose.;
CC pH dependence:
CC Optimum pH is 7.5. {ECO:0000269|PubMed:18392616};
CC Temperature dependence:
CC Optimum temperature is 30 degrees Celsius.
CC {ECO:0000269|PubMed:18392616};
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305|PubMed:17612504}.
CC -!- SIMILARITY: Belongs to the cellobiose 2-epimerase family.
CC {ECO:0000255|HAMAP-Rule:MF_00929}.
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DR EMBL; AB301953; BAF81108.1; -; Genomic_DNA.
DR PDB; 3VW5; X-ray; 2.60 A; A/B/C=1-389.
DR PDBsum; 3VW5; -.
DR AlphaFoldDB; P0DKY4; -.
DR SMR; P0DKY4; -.
DR MINT; P0DKY4; -.
DR KEGG; ag:BAF81108; -.
DR BioCyc; MetaCyc:MON-18526; -.
DR BRENDA; 5.1.3.11; 5477.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0047736; F:cellobiose epimerase activity; IEA:UniProtKB-UniRule.
DR GO; GO:0005975; P:carbohydrate metabolic process; IEA:InterPro.
DR Gene3D; 1.50.10.10; -; 1.
DR HAMAP; MF_00929; Cellobiose_2_epim; 1.
DR InterPro; IPR008928; 6-hairpin_glycosidase_sf.
DR InterPro; IPR012341; 6hp_glycosidase-like_sf.
DR InterPro; IPR028584; Cellobiose_2_epim.
DR SUPFAM; SSF48208; SSF48208; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cytoplasm; Direct protein sequencing; Isomerase.
FT CHAIN 1..389
FT /note="Cellobiose 2-epimerase"
FT /id="PRO_0000421448"
FT MUTAGEN 52
FT /note="R->A,K,H: Lack of activity."
FT /evidence="ECO:0000269|PubMed:19330485"
FT MUTAGEN 114
FT /note="F->A: Strong decrease in activity."
FT /evidence="ECO:0000269|PubMed:19330485"
FT MUTAGEN 243
FT /note="H->A,K,R: Lack of activity."
FT /evidence="ECO:0000269|PubMed:19330485"
FT MUTAGEN 246
FT /note="E->A,Q: Lack of activity."
FT /evidence="ECO:0000269|PubMed:19330485"
FT MUTAGEN 249
FT /note="W->F: Lack of activity."
FT /evidence="ECO:0000269|PubMed:19330485"
FT MUTAGEN 303
FT /note="W->F: Strong decrease in activity."
FT /evidence="ECO:0000269|PubMed:19330485"
FT MUTAGEN 304
FT /note="W->F: Lack of activity."
FT /evidence="ECO:0000269|PubMed:19330485"
FT MUTAGEN 308
FT /note="E->A,D,Q: Lack of activity."
FT /evidence="ECO:0000269|PubMed:19330485"
FT MUTAGEN 374
FT /note="H->A,K,R: Lack of activity."
FT /evidence="ECO:0000269|PubMed:19330485"
FT CONFLICT 3..8
FT /note="ISEIRQ -> KEEVKN (in Ref. 2; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 8
FT /note="Q -> N (in Ref. 1; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 12
FT /note="D -> E (in Ref. 1; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 12
FT /note="D -> S (in Ref. 2; AA sequence)"
FT /evidence="ECO:0000305"
FT HELIX 2..12
FT /evidence="ECO:0007829|PDB:3VW5"
FT HELIX 14..19
FT /evidence="ECO:0007829|PDB:3VW5"
FT TURN 24..26
FT /evidence="ECO:0007829|PDB:3VW5"
FT STRAND 27..29
FT /evidence="ECO:0007829|PDB:3VW5"
FT HELIX 47..64
FT /evidence="ECO:0007829|PDB:3VW5"
FT HELIX 67..83
FT /evidence="ECO:0007829|PDB:3VW5"
FT TURN 87..89
FT /evidence="ECO:0007829|PDB:3VW5"
FT STRAND 94..96
FT /evidence="ECO:0007829|PDB:3VW5"
FT STRAND 102..104
FT /evidence="ECO:0007829|PDB:3VW5"
FT HELIX 109..126
FT /evidence="ECO:0007829|PDB:3VW5"
FT HELIX 129..145
FT /evidence="ECO:0007829|PDB:3VW5"
FT TURN 166..168
FT /evidence="ECO:0007829|PDB:3VW5"
FT STRAND 170..172
FT /evidence="ECO:0007829|PDB:3VW5"
FT STRAND 176..178
FT /evidence="ECO:0007829|PDB:3VW5"
FT HELIX 179..196
FT /evidence="ECO:0007829|PDB:3VW5"
FT HELIX 199..214
FT /evidence="ECO:0007829|PDB:3VW5"
FT TURN 219..221
FT /evidence="ECO:0007829|PDB:3VW5"
FT STRAND 238..240
FT /evidence="ECO:0007829|PDB:3VW5"
FT HELIX 241..258
FT /evidence="ECO:0007829|PDB:3VW5"
FT HELIX 261..281
FT /evidence="ECO:0007829|PDB:3VW5"
FT STRAND 291..293
FT /evidence="ECO:0007829|PDB:3VW5"
FT STRAND 296..298
FT /evidence="ECO:0007829|PDB:3VW5"
FT HELIX 303..318
FT /evidence="ECO:0007829|PDB:3VW5"
FT HELIX 323..339
FT /evidence="ECO:0007829|PDB:3VW5"
FT STRAND 351..353
FT /evidence="ECO:0007829|PDB:3VW5"
FT STRAND 365..367
FT /evidence="ECO:0007829|PDB:3VW5"
FT HELIX 373..385
FT /evidence="ECO:0007829|PDB:3VW5"
SQ SEQUENCE 389 AA; 45220 MW; FC3DADD67BFA6433 CRC64;
MMISEIRQEL TDHIIPFWNK LRDDENGGFY GYLSYGLGLD KKADKGVILH SRILWFYSNA
YMTLGGDELL DNAKHAYEFI KNNCIDYEYG GVYWMMDFEG KPADTMKHTY NIAFAIYALS
SYYRASGDKE ALALAYRPFE DIEKNTLYEY GYREAFDRQW RLVDNEALSE NGLKADKTMN
AILHLIEAYT ELYKADGNEK VADRLKFQLG QMRDIVYTPD TNALKVFFDT AFNLVGDIHS
YGHDIEATWL MDRACDVLGD EDLKKQFAEM DLKISHNIQD IALEDGALNN ERDKNEIDKT
RVWWVQAEAV VGFINAYQHS GDEKFLESAK SVWENIKEYI IDKREGGEWY SEVTFDHTPH
DYKETVGPWK CPYHNGRMCM EVITRGVDI
