CDAS_BACSP
ID CDAS_BACSP Reviewed; 558 AA.
AC Q59226;
DT 23-FEB-2022, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1996, sequence version 1.
DT 03-AUG-2022, entry version 94.
DE RecName: Full=Cyclomaltodextrinase {ECO:0000303|PubMed:10620329, ECO:0000303|PubMed:11923309, ECO:0000303|PubMed:16536613, ECO:0000303|PubMed:9606956, ECO:0000312|EMBL:AAA92925.1};
DE Short=CDase {ECO:0000303|PubMed:10620329, ECO:0000303|PubMed:11923309, ECO:0000303|PubMed:16536613, ECO:0000303|PubMed:9606956};
DE Short=CDase I-5 {ECO:0000303|PubMed:10620329, ECO:0000303|PubMed:16536613, ECO:0000303|PubMed:9606956};
DE EC=3.2.1.135 {ECO:0000269|PubMed:9606956};
DE EC=3.2.1.54 {ECO:0000269|PubMed:10620329, ECO:0000269|PubMed:11923309, ECO:0000269|PubMed:16536613, ECO:0000269|PubMed:9606956, ECO:0000312|EMBL:AAA92925.1};
DE AltName: Full=Cyclomaltodextrin hydrolase, decycling {ECO:0000303|PubMed:10620329, ECO:0000303|PubMed:9606956};
GN Name=CDI5 {ECO:0000312|EMBL:AAA92925.1};
OS Bacillus sp.
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX NCBI_TaxID=1409 {ECO:0000312|EMBL:AAA92925.1};
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-7, FUNCTION,
RP CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, BIOTECHNOLOGY, AND MUTAGENESIS OF
RP VAL-380 AND ILE-388.
RC STRAIN=I-5 {ECO:0000303|PubMed:9606956, ECO:0000312|EMBL:AAA92925.1};
RX PubMed=9606956; DOI=10.1006/abbi.1998.0639;
RA Kim T.J., Shin J.H., Oh J.H., Kim M.J., Lee S.B., Ryu S., Kwon K.,
RA Kim J.W., Choi E.H., Robyt J.F., Park K.H.;
RT "Analysis of the gene encoding cyclomaltodextrinase from alkalophilic
RT Bacillus sp. I-5 and characterization of enzymatic properties.";
RL Arch. Biochem. Biophys. 353:221-227(1998).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, BIOTECHNOLOGY, AND REACTION MECHANISM.
RC STRAIN=I-5 {ECO:0000303|PubMed:10620329};
RX PubMed=10620329; DOI=10.1006/abbi.1999.1471;
RA Kim M.J., Park W.S., Lee H.S., Kim T.J., Shin J.H., Yoo S.H., Cheong T.K.,
RA Ryu S., Kim J.C., Kim J.W., Moon T.W., Robyt J.F., Park K.H.;
RT "Kinetics and inhibition of cyclomaltodextrinase from alkalophilic Bacillus
RT sp. I-5.";
RL Arch. Biochem. Biophys. 373:110-115(2000).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND BIOTECHNOLOGY.
RC STRAIN=I-5 {ECO:0000303|PubMed:16536613};
RX PubMed=16536613; DOI=10.1021/jf051887r;
RA Auh J.H., Chae H.Y., Kim Y.R., Shim K.H., Yoo S.H., Park K.H.;
RT "Modification of rice starch by selective degradation of amylose using
RT alkalophilic Bacillus cyclomaltodextrinase.";
RL J. Agric. Food Chem. 54:2314-2319(2006).
RN [4] {ECO:0007744|PDB:1EA9}
RP X-RAY CRYSTALLOGRAPHY (3.20 ANGSTROMS) OF 1-541, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RC STRAIN=I-5 {ECO:0000303|PubMed:11923309};
RX PubMed=11923309; DOI=10.1074/jbc.m201623200;
RA Lee H.S., Kim M.S., Cho H.S., Kim J.I., Kim T.J., Choi J.H., Park C.,
RA Lee H.S., Oh B.H., Park K.H.;
RT "Cyclomaltodextrinase, neopullulanase, and maltogenic amylase are nearly
RT indistinguishable from each other.";
RL J. Biol. Chem. 277:21891-21897(2002).
CC -!- FUNCTION: Hydrolyzes alpha-, beta- and gamma-cyclodextrins and the
CC resulting linear maltodextrins, with the highest activity with beta-
CC cyclodextrin (cyclomaltoheptaose). Soluble starch is hydrolyzed slowly,
CC but it is nevertheless preferred over pullulan as a substrate. Is able
CC to hydrolyze amylose and amylopectin, with a very strong preference for
CC amylose, with maltose and glucose as the main products (PubMed:9606956,
CC PubMed:10620329, PubMed:16536613). Maltose and glucose are the main
CC hydrolysis products of cyclomaltodextrins, maltodextrins and starch,
CC whereas panose is the main hydrolysis product of pullulan. Acarbose is
CC partially hydrolyzed to glucose and pseudotrisaccharide. No activity
CC with maltose as substrate (PubMed:9606956, PubMed:10620329). Has
CC transglycosylating activity with high concentrations of maltotriose,
CC maltotetraose and starch (PubMed:9606956).
CC {ECO:0000269|PubMed:10620329, ECO:0000269|PubMed:16536613,
CC ECO:0000269|PubMed:9606956}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=cyclomaltodextrin + H2O = linear maltodextrin;
CC Xref=Rhea:RHEA:23980, Rhea:RHEA-COMP:14584, Rhea:RHEA-COMP:14707,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:17623, ChEBI:CHEBI:18398; EC=3.2.1.54;
CC Evidence={ECO:0000269|PubMed:10620329, ECO:0000269|PubMed:11923309,
CC ECO:0000269|PubMed:16536613, ECO:0000269|PubMed:9606956};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:23981;
CC Evidence={ECO:0000269|PubMed:10620329, ECO:0000269|PubMed:11923309,
CC ECO:0000269|PubMed:16536613, ECO:0000269|PubMed:9606956};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Hydrolysis of pullulan to panose (6-alpha-D-glucosylmaltose).;
CC EC=3.2.1.135; Evidence={ECO:0000269|PubMed:9606956};
CC -!- COFACTOR:
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC Evidence={ECO:0000250|UniProtKB:P38940};
CC Note=Binds 1 Ca(2+) ion per subunit. {ECO:0000250|UniProtKB:P38940};
CC -!- ACTIVITY REGULATION: Hydrolysis of beta-cyclodextrin is inhibited by
CC Cu(2+), Zn(2+) and Ag(+), and activated by Ca(2+), EGTA and EDTA.
CC Activity is increased over twofold in the presence of 5 mM EDTA
CC (PubMed:9606956). Competitively inhibited by acarbose and methyl 6-
CC amino-6-deoxy-alpha-D-glucopyranoside by reducing the rate of the ring
CC opening step of the reaction (PubMed:10620329).
CC {ECO:0000269|PubMed:10620329, ECO:0000269|PubMed:9606956}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.23 mM for alpha-cyclodextrin (at pH 7.0 and 50 degrees Celsius)
CC {ECO:0000269|PubMed:10620329};
CC KM=0.83 mM for beta-cyclodextrin (at pH 7.0 and 50 degrees Celsius)
CC {ECO:0000269|PubMed:10620329};
CC KM=0.92 mM for gamma-cyclodextrin (at pH 7.0 and 50 degrees Celsius)
CC {ECO:0000269|PubMed:10620329};
CC KM=3.01 mg/ml for soluble starch (at pH 7.0 and 50 degrees Celsius)
CC {ECO:0000269|PubMed:10620329};
CC KM=2.11 mg/ml for pullulan (at pH 7.0 and 50 degrees Celsius)
CC {ECO:0000269|PubMed:10620329};
CC KM=0.515 mg/ml for beta-cyclodextrin (at pH 7.0 and 50 degrees
CC Celsius) {ECO:0000269|PubMed:16536613};
CC KM=1.52 mg/ml for amylose {ECO:0000269|PubMed:16536613};
CC KM=55.15 mg/ml for amylopectin {ECO:0000269|PubMed:16536613};
CC KM=1.43 mM for beta-cyclodextrin (with dimeric form of the enzyme at
CC pH 6.0 and 50 degrees Celsius) {ECO:0000269|PubMed:11923309};
CC KM=0.70 mM for beta-cyclodextrin (with dodecameric form of the enzyme
CC at pH 7.0 and 50 degrees Celsius) {ECO:0000269|PubMed:11923309};
CC Vmax=320.5 umol/min/mg enzyme with beta-cyclodextrin as substrate (at
CC pH 7.5 and 50 degrees Celsius) {ECO:0000269|PubMed:11923309};
CC Note=kcat is 78.18 sec(-1) with beta-cyclodextrin as substrate. kcat
CC is 22.11 sec(-1) with amylose as substrate. kcat is 50.36 sec(-1)
CC with amylopectin as substrate (PubMed:16536613). kcat is 101 min(-1)
CC for dimeric form of the enzyme with beta-cyclodextrin as substrate.
CC kcat is 521 min(-1) for dodecameric form of the enzyme with beta-
CC cyclodextrin as substrate (PubMed:11923309).
CC {ECO:0000269|PubMed:11923309, ECO:0000269|PubMed:16536613};
CC pH dependence:
CC Optimum pH for hydrolysis of beta-cyclodextrin is approximately 7.5
CC (PubMed:9606956, PubMed:16536613). Optimum pH for hydrolysis of
CC soluble starch is 6.0 and 6.0-7.0 for hydrolysis of pullulan
CC (PubMed:9606956). {ECO:0000269|PubMed:16536613,
CC ECO:0000269|PubMed:9606956};
CC Temperature dependence:
CC Optimum temperature is 50 degrees Celsius for hydrolysis of beta-
CC cyclodextrin (PubMed:9606956, PubMed:16536613). It is elevated from
CC 40 to 50 degrees Celsius by addition of Ca(2+), and the thermal
CC activity is retained more than 80% at 60 degrees Celsius in the
CC presence of Ca(2+). The optimum temperature is between 40-50 degrees
CC Celsius for hydrolysis of soluble starch, and it is 50 degrees
CC Celsius for hydrolysis of pullulan (PubMed:9606956).
CC {ECO:0000269|PubMed:16536613, ECO:0000269|PubMed:9606956};
CC -!- SUBUNIT: Monomer (PubMed:9606956, PubMed:11923309). Depending on the pH
CC of the solution, exists as a monomer, a homodimer or as an assembly of
CC six homodimers forming a dodecamer, which is catalytically the most
CC efficient form of the enzyme (PubMed:11923309).
CC {ECO:0000269|PubMed:11923309, ECO:0000269|PubMed:9606956}.
CC -!- BIOTECHNOLOGY: Cyclomaltodextrins form inclusion complexes with many
CC water-insoluble compounds that have applications in the food and drug
CC industries. The capability of this enzyme to hydrolyze
CC cyclomaltodextrins is of importance for the release of substances from
CC these inclusion complexes (PubMed:9606956, PubMed:10620329). This
CC enzyme can be used to modify rice starch structure to produce starch,
CC which has a decreased amylose content, or it could be used to produce
CC starch, which is free of amylose altogether. Cooked rice treated with
CC this enzyme has a significantly slower retrogradation
CC (PubMed:16536613). {ECO:0000269|PubMed:16536613,
CC ECO:0000305|PubMed:10620329, ECO:0000305|PubMed:9606956}.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 13 family. {ECO:0000305}.
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DR EMBL; U49646; AAA92925.1; -; Genomic_DNA.
DR PDB; 1EA9; X-ray; 3.20 A; C/D=1-541.
DR PDBsum; 1EA9; -.
DR SMR; Q59226; -.
DR CAZy; CBM34; Carbohydrate-Binding Module Family 34.
DR CAZy; GH13; Glycoside Hydrolase Family 13.
DR KEGG; ag:AAA92925; -.
DR EvolutionaryTrace; Q59226; -.
DR GO; GO:0005509; F:calcium ion binding; ISS:UniProtKB.
DR GO; GO:0047798; F:cyclomaltodextrinase activity; IDA:UniProtKB.
DR GO; GO:0005536; F:glucose binding; ISS:UniProtKB.
DR GO; GO:0042802; F:identical protein binding; IDA:UniProtKB.
DR GO; GO:0042803; F:protein homodimerization activity; IDA:UniProtKB.
DR GO; GO:2000897; P:amylopectin catabolic process; IDA:UniProtKB.
DR GO; GO:0051678; P:pullulan catabolic process; IDA:UniProtKB.
DR GO; GO:0005983; P:starch catabolic process; IDA:UniProtKB.
DR CDD; cd02857; E_set_CDase_PDE_N; 1.
DR Gene3D; 2.60.40.10; -; 1.
DR Gene3D; 2.60.40.1180; -; 1.
DR Gene3D; 3.90.400.10; -; 1.
DR InterPro; IPR006047; Glyco_hydro_13_cat_dom.
DR InterPro; IPR004185; Glyco_hydro_13_lg-like_dom.
DR InterPro; IPR013780; Glyco_hydro_b.
DR InterPro; IPR017853; Glycoside_hydrolase_SF.
DR InterPro; IPR013783; Ig-like_fold.
DR InterPro; IPR032091; Malt_amylase_C.
DR InterPro; IPR045857; O16G_dom_2.
DR Pfam; PF00128; Alpha-amylase; 1.
DR Pfam; PF02903; Alpha-amylase_N; 1.
DR Pfam; PF16657; Malt_amylase_C; 1.
DR SMART; SM00642; Aamy; 1.
DR SUPFAM; SSF51445; SSF51445; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Calcium; Carbohydrate metabolism; Direct protein sequencing;
KW Glycosidase; Hydrolase; Metal-binding.
FT CHAIN 1..558
FT /note="Cyclomaltodextrinase"
FT /id="PRO_0000454653"
FT ACT_SITE 325
FT /note="Nucleophile"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT ACT_SITE 354
FT /note="Proton donor"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 143
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 168
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 170
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 243
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 323
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 420..421
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 465
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 469
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT SITE 421
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000250|UniProtKB:P13507"
FT MUTAGEN 380
FT /note="V->T: Decreased activity with beta-cyclodextrin as
FT substrate, but an increase in starch hydrolyzing activity
FT compared to the wild type. No effect in pullulan
FT hydrolyzing activity."
FT /evidence="ECO:0000269|PubMed:9606956"
FT MUTAGEN 388
FT /note="I->E: Decreased activity with starch as substrate,
FT but an increase in beta-cyclodextrin hydrolyzing activity
FT compared to the wild type. No effect in pullulan
FT hydrolyzing activity."
FT /evidence="ECO:0000269|PubMed:9606956"
FT HELIX 3..5
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 14..21
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 35..42
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 44..46
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 52..56
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 58..61
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 66..68
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 79..81
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 96..99
FT /evidence="ECO:0007829|PDB:1EA9"
FT HELIX 107..109
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 110..113
FT /evidence="ECO:0007829|PDB:1EA9"
FT TURN 118..120
FT /evidence="ECO:0007829|PDB:1EA9"
FT HELIX 127..130
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 158..160
FT /evidence="ECO:0007829|PDB:1EA9"
FT HELIX 171..176
FT /evidence="ECO:0007829|PDB:1EA9"
FT HELIX 178..184
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 187..191
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 198..201
FT /evidence="ECO:0007829|PDB:1EA9"
FT TURN 213..215
FT /evidence="ECO:0007829|PDB:1EA9"
FT HELIX 218..228
FT /evidence="ECO:0007829|PDB:1EA9"
FT TURN 229..232
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 234..238
FT /evidence="ECO:0007829|PDB:1EA9"
FT TURN 246..248
FT /evidence="ECO:0007829|PDB:1EA9"
FT HELIX 249..255
FT /evidence="ECO:0007829|PDB:1EA9"
FT TURN 256..260
FT /evidence="ECO:0007829|PDB:1EA9"
FT TURN 262..265
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 271..273
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 285..288
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 292..294
FT /evidence="ECO:0007829|PDB:1EA9"
FT HELIX 300..317
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 320..324
FT /evidence="ECO:0007829|PDB:1EA9"
FT HELIX 332..345
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 350..353
FT /evidence="ECO:0007829|PDB:1EA9"
FT TURN 360..362
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 364..367
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 369..372
FT /evidence="ECO:0007829|PDB:1EA9"
FT HELIX 374..383
FT /evidence="ECO:0007829|PDB:1EA9"
FT HELIX 391..400
FT /evidence="ECO:0007829|PDB:1EA9"
FT TURN 401..404
FT /evidence="ECO:0007829|PDB:1EA9"
FT HELIX 407..411
FT /evidence="ECO:0007829|PDB:1EA9"
FT HELIX 425..429
FT /evidence="ECO:0007829|PDB:1EA9"
FT HELIX 433..443
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 446..450
FT /evidence="ECO:0007829|PDB:1EA9"
FT HELIX 465..468
FT /evidence="ECO:0007829|PDB:1EA9"
FT HELIX 481..496
FT /evidence="ECO:0007829|PDB:1EA9"
FT HELIX 499..502
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 516..522
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 527..533
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 535..537
FT /evidence="ECO:0007829|PDB:1EA9"
FT STRAND 539..541
FT /evidence="ECO:0007829|PDB:1EA9"
SQ SEQUENCE 558 AA; 64888 MW; B2E1C684B4289740 CRC64;
MFLEAVYHRP RKNWSYAYNG TTVHLRIRTK KDDMTAVYAL AGDKYMWDHT MEYVPMTKLA
TDELFDYWEC EVTPPYRRVK YGFLLQQGHE KRWMTEYDFL TEPPRNPDRL FEYPFINPVD
VFQPPAWVKD AIFYQIFPER FANGDTRNDP EGTLPWGSAD PTPSCFFGGD LQGVIDHLDH
LSKLGVNAVY FTPLFKATTN HKYDTEDYFQ IDPQFGDKDT LKKLVDLCHE RGIRVLLDAV
FNHSGRTFPP FVDVLKNGEK SKYKDWFHIR SLPLEVVDGI PTYDTFAFEP LMPKLNTEHP
DVKEYLLKAA EYWIRETGID GWRLDVANEV SHQFWREFRR VVKQANPDAY ILGEVWHESS
IWLEGDQFDA VMNYPFTNAV LDFFIHQIAD AEKFSFMLGK QLAGYPRQAS EVMFNLLDSH
DTARLLTQAD GDKRKMKLAV LFQFTYFGTP CIYYGDEVGL DGGHDPGCRK CMEWDETKHD
KDLFAFYQTV IRLRQAHAAL RTGTFKFLTA EKNSRQIAYL REDDQDTILV VMNNDKAGHT
LRCLSGMHSG PICGTTMS
