Open AccessCrystal structure of endo‐1,4‐β‐glucanase from Eisenia fetida
Author(s) -
Arimori Takao,
Ito Akihiro,
Nakazawa Masami,
Ueda Mitsuhiro,
Tamada Taro
Publication year - 2013
Publication title -
journal of synchrotron radiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.172
H-Index - 99
ISSN - 1600-5775
DOI - 10.1107/s0909049513021110
Subject(s) - eisenia fetida , hydrolysis , pichia pastoris , hydrolase , glycoside hydrolase , chemistry , cellulase , glucanase , crystallization , fermentation , recombinant dna , enzyme , food science , biochemistry , organic chemistry , gene , toxicity
The saccharification process is essential for bioethanol production from woody biomass including celluloses. Cold‐adapted cellulase, which has sufficient activity at low temperature (<293 K), is capable of reducing heating costs during the saccharification process and is suitable for simultaneous saccharification and fermentation. Endo‐1,4‐β‐glucanase from the earthworm Eisenia fetida (EF‐EG2) belonging to glycoside hydrolase family 9 has been shown to have the highest activity at 313 K, and also retained a comparatively high activity at 283 K. The recombinant EF‐EG2 was purified expressed in Pichia pastoris, and then grew needle‐shaped crystals with dimensions of 0.02 × 0.02 × 1 mm. The crystals belonged to the space group P 3 2 21 with unit‐cell parameters of a = b = 136 Å, c = 55.0 Å. The final model of EF‐EG2, including 435 residues, two ions, seven crystallization reagents and 696 waters, was refined to a crystallographic R ‐factor of 14.7% (free R ‐factor of 16.8%) to 1.5 Å resolution. The overall structure of EF‐EG2 has an (α/α) 6 barrel fold which contains a putative active‐site cleft and a negatively charged surface. This structural information helps us understand the catalytic and cold adaptation mechanisms of EF‐EG2.