5Y6T

Crystal structure of endo-1,4-beta-mannanase from Eisenia fetida

5Y6T の概要

• エントリーDOI: 10.2210/pdb5y6t/pdb
• 分子名称: endo-1,4-beta-mannanase, 2-AMINO-2-HYDROXYMETHYL-PROPANE-1,3-DIOL, ISOPROPYL ALCOHOL, ... (4 entities in total)
• 機能のキーワード: cold adaptation, gh5 family, carbohydrate
• 由来する生物種: Eisenia fetida
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 42809.80

構造登録者

Hirano, Y.,Ueda, M.,Tamada, T. (登録日: 2017-08-15, 公開日: 2018-06-27, 最終更新日: 2024-10-23)

主引用文献

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

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: 30037547
DOI: 10.1016/j.enzmictec.2018.05.014

実験手法

X-RAY DIFFRACTION (1.7 Å)