A novel cold-adapted cellulase complex from Eisenia foetida: characterization of a multienzyme complex with carboxymethylcellulase, beta-glucosidase, beta-1,3 glucanase, and beta-xylosidase.
Clostridium sp. and some bacterial cellulases exist as an enzyme complex with cellulolytic, and hemicellulolytic enzymes, so called "cellulosome". In this article, we report that EF-CMCase25 occurs as a complex with beta-glucosidase, beta-1,3 glucanase, and beta-xylosidase. The multienzyme complex had a molecular mass of 150 kDa on gel filtration under non-reducing condition. After the gel filtration, the enzyme complex was purified to homogeneous state on BN-PAGE. The SDS-PAGE demonstrated that the purified protein is a complex with at least one CMCase (25 kDa), one beta-glucosidase (32 kDa), and one beta-1,3 glucanase (40 kDa) components. The CMCase activity in the purified enzyme complex at 15 degrees C was 44% of that obtained at the optimal temperature. The optimum pH of the EF-CMCase25 in the purified enzyme complex was pH 5.0 and stable at pH 7.0-9.0.
Ueda
Mitsuhiro
M
Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Sakai, Osaka, 599-8531, Japan. mueda@biochem.osakafu-u.ac.jp
Goto
Takahiro
T
Nakazawa
Masami
M
Miyatake
Kazutaka
K
Sakaguchi
Minoru
M
Inouye
Kuniyo
K
D000222:Adaptation, Physiological; D000818:Animals; D002480:Cellulase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D003080:Cold Temperature; D004795:Enzyme Stability; D043326:Glucan 1,3-beta-Glucosidase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D009835:Oligochaeta / Q000201:enzymology* / Q000502:physiology*; D013379:Substrate Specificity; D013696:Temperature; D014995:Xylosidases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D001617:beta-Glucosidase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
D009097:Multienzyme Complexes; D014995:Xylosidases; D001617:beta-Glucosidase; C026579:exo-1,4-beta-D-xylosidase; D002480:Cellulase; C019826:carboxymethylcellulase; D043326:Glucan 1,3-beta-Glucosidase