5Y6T
Crystal structure of endo-1,4-beta-mannanase from Eisenia fetida
Summary for 5Y6T
| Entry DOI | 10.2210/pdb5y6t/pdb |
| Descriptor | endo-1,4-beta-mannanase, 2-AMINO-2-HYDROXYMETHYL-PROPANE-1,3-DIOL, ISOPROPYL ALCOHOL, ... (4 entities in total) |
| Functional Keywords | cold adaptation, gh5 family, carbohydrate |
| Biological source | Eisenia fetida |
| Total number of polymer chains | 1 |
| Total formula weight | 42809.80 |
| Authors | Hirano, Y.,Ueda, M.,Tamada, T. (deposition date: 2017-08-15, release date: 2018-06-27, Last modification date: 2024-10-23) |
| Primary citation | Ueda, M.,Hirano, Y.,Fukuhara, H.,Naka, Y.,Nakazawa, M.,Sakamoto, T.,Ogata, Y.,Tamada, T. Gene cloning, expression, and X-ray crystallographic analysis of a beta-mannanase from Eisenia fetida. Enzyme.Microb.Technol., 117:15-22, 2018 Cited by PubMed Abstract: The endo-1,4-β-mannanases (Ef-Man) gene from Eisenia fetida was determined to consist of 1131 bp and encode a 377 amino acid protein. The amino acid sequence showed similarity with the endo-1,4-β-mannanases of Daphnia pulex (62%), Cryptopygus antarcticus (64%), Crassostrea gigas (61%), Mytilus edulis (60%), and Aplysia kurodai (58%). The gene encoding mature Ef-Man was expressed in Pichia pastoris (GS115 strain). Based on SDS-PAGE analysis, the molecular mass of the purified recombinant Ef-Man (rEf-Man) was estimated to be 39 kDa. All catalytically important residues of endo-1,4-β-mannanases in the glycoside hydrolase (GH) family 5 were conserved in Ef-Man. The optimal temperature for rEf-Man was identified as 60 °C. HPLC and HPAEC analyses suggest that Ef-Man requires at least six subsites for efficient hydrolysis and is capable of performing transglycosylation reactions. The overall structure of rEf-Man is similar to those of GH5 family proteins, and tertiary structures around the active site are conserved among endo-1,4-β-mannanase families. X-ray crystallographic analysis supports the hydrolysis and transglycosylation reaction mechanism determined by HPLC and HPAEC analyses. PubMed: 30037547DOI: 10.1016/j.enzmictec.2018.05.014 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.7 Å) |
Structure validation
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