| IED ID | IndEnz0001000315 |
| Enzyme Type ID | amylase000315 |
| Protein Name |
Cyclomaltodextrinase CDase CDase I-5 EC 3.2.1.135 EC 3.2.1.54 Cyclomaltodextrin hydrolase, decycling |
| Gene Name | CDI5 |
| Organism | Bacillus sp. |
| Taxonomic Lineage | cellular organisms Bacteria Terrabacteria group Firmicutes Bacilli Bacillales Bacillaceae Bacillus unclassified Bacillus (in: Bacteria) Bacillus sp. |
| Enzyme Sequence | MFLEAVYHRPRKNWSYAYNGTTVHLRIRTKKDDMTAVYALAGDKYMWDHTMEYVPMTKLATDELFDYWECEVTPPYRRVKYGFLLQQGHEKRWMTEYDFLTEPPRNPDRLFEYPFINPVDVFQPPAWVKDAIFYQIFPERFANGDTRNDPEGTLPWGSADPTPSCFFGGDLQGVIDHLDHLSKLGVNAVYFTPLFKATTNHKYDTEDYFQIDPQFGDKDTLKKLVDLCHERGIRVLLDAVFNHSGRTFPPFVDVLKNGEKSKYKDWFHIRSLPLEVVDGIPTYDTFAFEPLMPKLNTEHPDVKEYLLKAAEYWIRETGIDGWRLDVANEVSHQFWREFRRVVKQANPDAYILGEVWHESSIWLEGDQFDAVMNYPFTNAVLDFFIHQIADAEKFSFMLGKQLAGYPRQASEVMFNLLDSHDTARLLTQADGDKRKMKLAVLFQFTYFGTPCIYYGDEVGLDGGHDPGCRKCMEWDETKHDKDLFAFYQTVIRLRQAHAALRTGTFKFLTAEKNSRQIAYLREDDQDTILVVMNNDKAGHTLRCLSGMHSGPICGTTMS |
| Enzyme Length | 558 |
| Uniprot Accession Number | Q59226 |
| Absorption | |
| Active Site | ACT_SITE 325; /note=Nucleophile; /evidence=ECO:0000250|UniProtKB:P38940; ACT_SITE 354; /note=Proton donor; /evidence=ECO:0000250|UniProtKB:P38940 |
| Activity Regulation | ACTIVITY REGULATION: Hydrolysis of beta-cyclodextrin is inhibited by Cu(2+), Zn(2+) and Ag(+), and activated by Ca(2+), EGTA and EDTA. Activity is increased over twofold in the presence of 5 mM EDTA (PubMed:9606956). Competitively inhibited by acarbose and methyl 6-amino-6-deoxy-alpha-D-glucopyranoside by reducing the rate of the ring opening step of the reaction (PubMed:10620329). {ECO:0000269|PubMed:10620329, ECO:0000269|PubMed:9606956}. |
| Binding Site | BINDING 243; /note=Substrate; /evidence=ECO:0000250|UniProtKB:P38940; BINDING 323; /note=Substrate; /evidence=ECO:0000250|UniProtKB:P38940; BINDING 465; /note=Substrate; /evidence=ECO:0000250|UniProtKB:P38940; BINDING 469; /note=Substrate; /evidence=ECO:0000250|UniProtKB:P38940 |
| Calcium Binding | |
| catalytic Activity | CATALYTIC ACTIVITY: Reaction=cyclomaltodextrin + H2O = linear maltodextrin; Xref=Rhea:RHEA:23980, Rhea:RHEA-COMP:14584, Rhea:RHEA-COMP:14707, ChEBI:CHEBI:15377, ChEBI:CHEBI:17623, ChEBI:CHEBI:18398; EC=3.2.1.54; Evidence={ECO:0000269|PubMed:10620329, ECO:0000269|PubMed:11923309, ECO:0000269|PubMed:16536613, ECO:0000269|PubMed:9606956};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:23981; Evidence={ECO:0000269|PubMed:10620329, ECO:0000269|PubMed:11923309, ECO:0000269|PubMed:16536613, ECO:0000269|PubMed:9606956}; CATALYTIC ACTIVITY: Reaction=Hydrolysis of pullulan to panose (6-alpha-D-glucosylmaltose).; EC=3.2.1.135; Evidence={ECO:0000269|PubMed:9606956}; |
| DNA Binding | |
| EC Number | 3.2.1.135; 3.2.1.54 |
| Enzyme Function | FUNCTION: Hydrolyzes alpha-, beta- and gamma-cyclodextrins and the resulting linear maltodextrins, with the highest activity with beta-cyclodextrin (cyclomaltoheptaose). Soluble starch is hydrolyzed slowly, but it is nevertheless preferred over pullulan as a substrate. Is able to hydrolyze amylose and amylopectin, with a very strong preference for amylose, with maltose and glucose as the main products (PubMed:9606956, PubMed:10620329, PubMed:16536613). Maltose and glucose are the main hydrolysis products of cyclomaltodextrins, maltodextrins and starch, whereas panose is the main hydrolysis product of pullulan. Acarbose is partially hydrolyzed to glucose and pseudotrisaccharide. No activity with maltose as substrate (PubMed:9606956, PubMed:10620329). Has transglycosylating activity with high concentrations of maltotriose, maltotetraose and starch (PubMed:9606956). {ECO:0000269|PubMed:10620329, ECO:0000269|PubMed:16536613, ECO:0000269|PubMed:9606956}. |
| Temperature Dependency | BIOPHYSICOCHEMICAL PROPERTIES: Temperature dependence: Optimum temperature is 50 degrees Celsius for hydrolysis of beta-cyclodextrin (PubMed:9606956, PubMed:16536613). It is elevated from 40 to 50 degrees Celsius by addition of Ca(2+), and the thermal activity is retained more than 80% at 60 degrees Celsius in the presence of Ca(2+). The optimum temperature is between 40-50 degrees Celsius for hydrolysis of soluble starch, and it is 50 degrees Celsius for hydrolysis of pullulan (PubMed:9606956). {ECO:0000269|PubMed:16536613, ECO:0000269|PubMed:9606956}; |
| PH Dependency | BIOPHYSICOCHEMICAL PROPERTIES: pH dependence: Optimum pH for hydrolysis of beta-cyclodextrin is approximately 7.5 (PubMed:9606956, PubMed:16536613). Optimum pH for hydrolysis of soluble starch is 6.0 and 6.0-7.0 for hydrolysis of pullulan (PubMed:9606956). {ECO:0000269|PubMed:16536613, ECO:0000269|PubMed:9606956}; |
| Pathway | |
| nucleotide Binding | |
| Features | Active site (2); Binding site (4); Chain (1); Metal binding (3); Mutagenesis (2); Region (1); Site (1) |
| Keywords | 3D-structure;Calcium;Carbohydrate metabolism;Direct protein sequencing;Glycosidase;Hydrolase;Metal-binding |
| Interact With | |
| Induction | |
| Subcellular Location | |
| Modified Residue | |
| Post Translational Modification | |
| Signal Peptide | |
| Structure 3D | X-ray crystallography (1) |
| Cross Reference PDB | 1EA9; |
| Mapped Pubmed ID | - |
| Motif | |
| Gene Encoded By | |
| Mass | 64,888 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=1.23 mM for alpha-cyclodextrin (at pH 7.0 and 50 degrees Celsius) {ECO:0000269|PubMed:10620329}; KM=0.83 mM for beta-cyclodextrin (at pH 7.0 and 50 degrees Celsius) {ECO:0000269|PubMed:10620329}; KM=0.92 mM for gamma-cyclodextrin (at pH 7.0 and 50 degrees Celsius) {ECO:0000269|PubMed:10620329}; KM=3.01 mg/ml for soluble starch (at pH 7.0 and 50 degrees Celsius) {ECO:0000269|PubMed:10620329}; KM=2.11 mg/ml for pullulan (at pH 7.0 and 50 degrees Celsius) {ECO:0000269|PubMed:10620329}; KM=0.515 mg/ml for beta-cyclodextrin (at pH 7.0 and 50 degrees Celsius) {ECO:0000269|PubMed:16536613}; KM=1.52 mg/ml for amylose {ECO:0000269|PubMed:16536613}; KM=55.15 mg/ml for amylopectin {ECO:0000269|PubMed:16536613}; KM=1.43 mM for beta-cyclodextrin (with dimeric form of the enzyme at pH 6.0 and 50 degrees Celsius) {ECO:0000269|PubMed:11923309}; KM=0.70 mM for beta-cyclodextrin (with dodecameric form of the enzyme at pH 7.0 and 50 degrees Celsius) {ECO:0000269|PubMed:11923309}; Vmax=320.5 umol/min/mg enzyme with beta-cyclodextrin as substrate (at pH 7.5 and 50 degrees Celsius) {ECO:0000269|PubMed:11923309}; Note=kcat is 78.18 sec(-1) with beta-cyclodextrin as substrate. kcat is 22.11 sec(-1) with amylose as substrate. kcat is 50.36 sec(-1) with amylopectin as substrate (PubMed:16536613). kcat is 101 min(-1) for dimeric form of the enzyme with beta-cyclodextrin as substrate. kcat is 521 min(-1) for dodecameric form of the enzyme with beta-cyclodextrin as substrate (PubMed:11923309). {ECO:0000269|PubMed:11923309, ECO:0000269|PubMed:16536613}; |
| Metal Binding | METAL 143; /note=Calcium; /evidence=ECO:0000250|UniProtKB:P38940; METAL 168; /note=Calcium; via carbonyl oxygen; /evidence=ECO:0000250|UniProtKB:P38940; METAL 170; /note=Calcium; /evidence=ECO:0000250|UniProtKB:P38940 |
| Rhea ID | RHEA:23980; RHEA:23981 |
| Cross Reference Brenda |