Structural highlights
FunctionPublication Abstract from PubMedThe endo-1,4-beta-mannanases (Ef-Man) gene from Eisenia fetida was determined to consist of 1131bp and encode a 377 amino acid protein. The amino acid sequence showed similarity with the endo-1,4-beta-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 39kDa. All catalytically important residues of endo-1,4-beta-mannanases in the glycoside hydrolase (GH) family 5 were conserved in Ef-Man. The optimal temperature for rEf-Man was identified as 60 degrees 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-beta-mannanase families. X-ray crystallographic analysis supports the hydrolysis and transglycosylation reaction mechanism determined by HPLC and HPAEC analyses. Gene cloning, expression, and X-ray crystallographic analysis of a beta-mannanase from Eisenia fetida.,Ueda M, Hirano Y, Fukuhara H, Naka Y, Nakazawa M, Sakamoto T, Ogata Y, Tamada T Enzyme Microb Technol. 2018 Oct;117:15-22. doi: 10.1016/j.enzmictec.2018.05.014. , Epub 2018 May 25. PMID:30037547[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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