| IED ID | IndEnz0001000120 |
| Enzyme Type ID | amylase000120 |
| Protein Name |
Cyclomaltodextrinase CDase EC 3.2.1.54 Cyclomaltodextrin hydrolase, decycling |
| Gene Name | T260_08735 |
| Organism | Geobacillus thermopakistaniensis (strain MAS1) |
| Taxonomic Lineage | cellular organisms Bacteria Terrabacteria group Firmicutes Bacilli Bacillales Bacillaceae Geobacillus unclassified Geobacillus Geobacillus thermopakistaniensis (strain MAS1) |
| Enzyme Sequence | MRKEAIHHRSTDNFAYAYDSETLHLRLQTKKNDVDHVELLFGDPYEWHDGAWQFQTMPMRKTGSDGLFDYWLAEVKPPYRRLRYGFVLRAGGEKLVYTEKGFYHEAPSDDTAYYFCFPFLHRVDLFQAPDWVKDTVWYQIFPERFANGNPAISPKGARPWGSEDPTPTSFFGGDLQGIIDHLDYLADLGITGIYLTPIFRAPSNHKYDTADYFEIDPHFGDKETLKTLVKRCHEKGIRVMLDAVFNHCGYEFGPFQDVLKNGAASRYKDWFHIREFPLQTEPRPNYDTFAFVPQMPKLNTAHPEVKRYLLDVATYWIREFDIDGWRLDVANEIDHQFWREFRQAVKALKPDVYILGEIWHDAMPWLRGDQFDAVMNYPLADAALRFFAKEDMSASEFADRLMHVLHSYPKQVNEAAFNLLGSHDTPRLLTVCGGDVRKVKLLFLFQLTFTGSPCIYYGDEIGMTGGNDPECRKCMVWDPEKQNKELYEHVKQLIALRKQYRALRRGDVAFLAADDEVNHLVYAKTDGNETVMIIINRSNEAAEIPMPIDARGKWLVNLLTGERFAAEAETLCVSLPPYGFVLYAVESW |
| Enzyme Length | 588 |
| Uniprot Accession Number | A0A7U9P668 |
| Absorption | |
| Active Site | ACT_SITE 328; /note=Nucleophile; /evidence=ECO:0000250|UniProtKB:P38940; ACT_SITE 357; /note=Proton donor; /evidence=ECO:0000250|UniProtKB:P38940 |
| Activity Regulation | ACTIVITY REGULATION: No metal dependence, but Mn(2+) increases the activity with alpha-cyclodextrin as substrate. No effect on the activity with presence or absence of Ca(2+), Zn(2+), Tween-20 or EDTA. {ECO:0000269|PubMed:31212108}. |
| Binding Site | BINDING 247; /note=Substrate; /evidence=ECO:0000250|UniProtKB:P38940; BINDING 326; /note=Substrate; /evidence=ECO:0000250|UniProtKB:P38940; BINDING 468; /note=Substrate; /evidence=ECO:0000250|UniProtKB:P38940; BINDING 472; /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:31212108};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:23981; Evidence={ECO:0000269|PubMed:31212108}; |
| DNA Binding | |
| EC Number | 3.2.1.54 |
| Enzyme Function | FUNCTION: Hydrolyzes alpha-, beta- and gamma-cyclodextrins with the highest activity with alpha-cyclodextrin (cyclomaltohexaose). Pullulan is the preferred substrate from linear substrates. Maltose is a major product of these reactions. Is also able to hydrolyze maltotriose and acarbose, and transglycosylate their hydrolytic products. Major reaction products of maltotriose and of acarbose are maltose and glucose, and glucose and pseudotrisaccharide, respectively. No activity with glucose or maltose as substrate. {ECO:0000269|PubMed:31212108}. |
| Temperature Dependency | BIOPHYSICOCHEMICAL PROPERTIES: Temperature dependence: Optimum temperature is 55 degrees Celsius for hydrolysis of alpha-cyclodextrin. Thermostable. No significant loss of catalytic activity even after overnight incubation at 65 degrees. EDTA enhances the thermostability, the half-life being 90 min at 70 degrees Celsius in the absence of EDTA, increasing to 360 min in the presence of 10 mM EDTA. However, overnight incubation at 4 degrees Celsius is first required for enhancement of thermostability. Activity decreases drastically at 75 degrees Celsius. {ECO:0000269|PubMed:31212108}; |
| PH Dependency | BIOPHYSICOCHEMICAL PROPERTIES: pH dependence: Optimum pH is 6.0 for hydrolysis of alpha-cyclodextrin. {ECO:0000269|PubMed:31212108}; |
| Pathway | |
| nucleotide Binding | |
| Features | Active site (2); Binding site (4); Chain (1); Region (1); Site (1) |
| Keywords | Carbohydrate metabolism;Glycosidase;Hydrolase |
| Interact With | |
| Induction | |
| Subcellular Location | |
| Modified Residue | |
| Post Translational Modification | |
| Signal Peptide | |
| Structure 3D | |
| Cross Reference PDB | - |
| Mapped Pubmed ID | 31212108; |
| Motif | |
| Gene Encoded By | |
| Mass | 68,157 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=0.8 mM for alpha-cyclodextrin {ECO:0000269|PubMed:31212108}; KM=0.6 mM for beta-cyclodextrin {ECO:0000269|PubMed:31212108}; KM=0.4 mM for gamma-cyclodextrin {ECO:0000269|PubMed:31212108}; Vmax=1200 umol/min/mg enzyme with alpha-cyclodextrin as substrate {ECO:0000269|PubMed:31212108}; Vmax=735 umol/min/mg enzyme with beta-cyclodextrin as substrate {ECO:0000269|PubMed:31212108}; Vmax=360 umol/min/mg enzyme with gamma-cyclodextrin as substrate {ECO:0000269|PubMed:31212108}; Vmax=105 umol/min/mg enzyme with pullulan as substrate {ECO:0000269|PubMed:31212108}; Note=kcat is 2739 sec(-1) with alpha-cyclodextrin as substrate. kcat is 1678 sec(-1) with beta-cyclodextrin as substrate. kcat is 821 sec(-1) with gamma-cyclodextrin as substrate. kcat value calculations are based on the dimeric enzyme. {ECO:0000269|PubMed:31212108}; |
| Metal Binding | |
| Rhea ID | RHEA:23980; RHEA:23981 |
| Cross Reference Brenda |