CDAS_GEOTM
ID CDAS_GEOTM Reviewed; 588 AA.
AC A0A7U9P668;
DT 23-FEB-2022, integrated into UniProtKB/Swiss-Prot.
DT 02-JUN-2021, sequence version 1.
DT 03-AUG-2022, entry version 4.
DE RecName: Full=Cyclomaltodextrinase {ECO:0000303|PubMed:31212108, ECO:0000312|EMBL:ESU72342.1};
DE Short=CDase {ECO:0000303|PubMed:31212108};
DE EC=3.2.1.54 {ECO:0000269|PubMed:31212108};
DE AltName: Full=Cyclomaltodextrin hydrolase, decycling {ECO:0000305};
GN ORFNames=T260_08735 {ECO:0000312|EMBL:ESU72342.1};
OS Geobacillus thermopakistaniensis (strain MAS1).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Geobacillus;
OC unclassified Geobacillus.
OX NCBI_TaxID=1408282;
RN [1] {ECO:0000312|EMBL:ESU72342.1, ECO:0000312|Proteomes:UP000018339}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=MAS1 {ECO:0000312|EMBL:ESU72342.1,
RC ECO:0000312|Proteomes:UP000018339};
RX PubMed=24903880; DOI=10.1128/genomea.00559-14;
RA Siddiqui M.A., Rashid N., Ayyampalayam S., Whitman W.B.;
RT "Draft Genome Sequence of Geobacillus thermopakistaniensis Strain MAS1.";
RL Genome Announc. 2:E0055914-E0055914(2014).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RX PubMed=31212108; DOI=10.1016/j.carres.2019.06.004;
RA Aroob I., Ahmad N., Aslam M., Shaeer A., Rashid N.;
RT "A highly active alpha-cyclodextrin preferring cyclomaltodextrinase from
RT Geobacillus thermopakistaniensis.";
RL Carbohydr. Res. 481:1-8(2019).
CC -!- FUNCTION: Hydrolyzes alpha-, beta- and gamma-cyclodextrins with the
CC highest activity with alpha-cyclodextrin (cyclomaltohexaose). Pullulan
CC is the preferred substrate from linear substrates. Maltose is a major
CC product of these reactions. Is also able to hydrolyze maltotriose and
CC acarbose, and transglycosylate their hydrolytic products. Major
CC reaction products of maltotriose and of acarbose are maltose and
CC glucose, and glucose and pseudotrisaccharide, respectively. No activity
CC with glucose or maltose as substrate. {ECO:0000269|PubMed:31212108}.
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:31212108};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:23981;
CC Evidence={ECO:0000269|PubMed:31212108};
CC -!- ACTIVITY REGULATION: No metal dependence, but Mn(2+) increases the
CC activity with alpha-cyclodextrin as substrate. No effect on the
CC activity with presence or absence of Ca(2+), Zn(2+), Tween-20 or EDTA.
CC {ECO:0000269|PubMed:31212108}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.8 mM for alpha-cyclodextrin {ECO:0000269|PubMed:31212108};
CC KM=0.6 mM for beta-cyclodextrin {ECO:0000269|PubMed:31212108};
CC KM=0.4 mM for gamma-cyclodextrin {ECO:0000269|PubMed:31212108};
CC Vmax=1200 umol/min/mg enzyme with alpha-cyclodextrin as substrate
CC {ECO:0000269|PubMed:31212108};
CC Vmax=735 umol/min/mg enzyme with beta-cyclodextrin as substrate
CC {ECO:0000269|PubMed:31212108};
CC Vmax=360 umol/min/mg enzyme with gamma-cyclodextrin as substrate
CC {ECO:0000269|PubMed:31212108};
CC Vmax=105 umol/min/mg enzyme with pullulan as substrate
CC {ECO:0000269|PubMed:31212108};
CC Note=kcat is 2739 sec(-1) with alpha-cyclodextrin as substrate. kcat
CC is 1678 sec(-1) with beta-cyclodextrin as substrate. kcat is 821
CC sec(-1) with gamma-cyclodextrin as substrate. kcat value calculations
CC are based on the dimeric enzyme. {ECO:0000269|PubMed:31212108};
CC pH dependence:
CC Optimum pH is 6.0 for hydrolysis of alpha-cyclodextrin.
CC {ECO:0000269|PubMed:31212108};
CC Temperature dependence:
CC Optimum temperature is 55 degrees Celsius for hydrolysis of alpha-
CC cyclodextrin. Thermostable. No significant loss of catalytic activity
CC even after overnight incubation at 65 degrees. EDTA enhances the
CC thermostability, the half-life being 90 min at 70 degrees Celsius in
CC the absence of EDTA, increasing to 360 min in the presence of 10 mM
CC EDTA. However, overnight incubation at 4 degrees Celsius is first
CC required for enhancement of thermostability. Activity decreases
CC drastically at 75 degrees Celsius. {ECO:0000269|PubMed:31212108};
CC -!- SUBUNIT: Exists as a monomer or a homodimer in solution. Homodimer is
CC more active and stable than the monomer. {ECO:0000269|PubMed:31212108}.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 13 family. {ECO:0000305}.
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DR EMBL; AYSF01000045; ESU72342.1; -; Genomic_DNA.
DR RefSeq; WP_023633943.1; NZ_AYSF01000045.1.
DR SMR; A0A7U9P668; -.
DR Proteomes; UP000018339; Unassembled WGS sequence.
DR GO; GO:0047798; F:cyclomaltodextrinase activity; IDA:UniProtKB.
DR GO; GO:0042802; F:identical protein binding; IDA:UniProtKB.
DR GO; GO:0042803; F:protein homodimerization activity; IDA:UniProtKB.
DR GO; GO:0051678; P:pullulan 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 Carbohydrate metabolism; Glycosidase; Hydrolase.
FT CHAIN 1..588
FT /note="Cyclomaltodextrinase"
FT /id="PRO_0000454654"
FT ACT_SITE 328
FT /note="Nucleophile"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT ACT_SITE 357
FT /note="Proton donor"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 247
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 326
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 423..424
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 468
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 472
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT SITE 424
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000250|UniProtKB:P13507"
SQ SEQUENCE 588 AA; 68157 MW; 72E0837B6575C1A8 CRC64;
MRKEAIHHRS TDNFAYAYDS ETLHLRLQTK KNDVDHVELL FGDPYEWHDG AWQFQTMPMR
KTGSDGLFDY WLAEVKPPYR RLRYGFVLRA GGEKLVYTEK GFYHEAPSDD TAYYFCFPFL
HRVDLFQAPD WVKDTVWYQI FPERFANGNP AISPKGARPW GSEDPTPTSF FGGDLQGIID
HLDYLADLGI TGIYLTPIFR APSNHKYDTA DYFEIDPHFG DKETLKTLVK RCHEKGIRVM
LDAVFNHCGY EFGPFQDVLK NGAASRYKDW FHIREFPLQT EPRPNYDTFA FVPQMPKLNT
AHPEVKRYLL DVATYWIREF DIDGWRLDVA NEIDHQFWRE FRQAVKALKP DVYILGEIWH
DAMPWLRGDQ FDAVMNYPLA DAALRFFAKE DMSASEFADR LMHVLHSYPK QVNEAAFNLL
GSHDTPRLLT VCGGDVRKVK LLFLFQLTFT GSPCIYYGDE IGMTGGNDPE CRKCMVWDPE
KQNKELYEHV KQLIALRKQY RALRRGDVAF LAADDEVNHL VYAKTDGNET VMIIINRSNE
AAEIPMPIDA RGKWLVNLLT GERFAAEAET LCVSLPPYGF VLYAVESW
