@article{oai:repo.qst.go.jp:00049415,
 author = {上田, 光宏 and 平野, 優 and 福原, 宏章 and 中, 裕規 and 中澤, 昌美 and 阪本, 龍司 and Ogata, Yoshiyuki and 玉田, 太郎 and 平野 優 and 玉田 太郎},
 journal = {Enzyme and Microbial Technology},
 month = {May},
 note = {The Ef-Man gene 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-beta-mannanases of Daphnia pulex (62%), Cryptopygus antarcticus (64%), Crassostrea gigas (61%),  Mytilus edulis (60%), and Aplisia 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 39.5 kDa. 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°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.},
 pages = {15--22},
 title = {Gene cloning, expression, and X-ray crystallographic analysis of a β-mannanase from Eisenia fetida},
 volume = {117},
 year = {2018}
}