Isolation of cellulolytic enzymes from moldy silage by new culture-independent strategy.
A culture-independent strategy has been developed for investigation of cellulases in moldy silage. By the qualitative differences in the adsorption of cellulases on lingo-cellulosics, a new cellobiohydrolase (CBH) with apparent molecular mass of 194 kDa was isolated and characterized. The entire extracellular proteins of silage were separated by two-dimensional gel electrophoresis, and five potential endoglucanases were identified by activity staining. These results demonstrate the feasibility of direct screening cellulases from environment without microorganism cultivation and this strategy could be expected to facilitate the research of uncultured microorganisms.
Yu
Rentao
R
State Key Laboratory of Microbial Technology, Shandong University, Jinan, China.
Wang
Lushan
L
Duan
Xinyuan
X
Gao
Peiji
P
D001709:Biotechnology / Q000379:methods*; D043366:Cellulose 1,4-beta-Cellobiosidase / Q000737:chemistry / Q000302:isolation & purification*; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D040901:Proteomics; D012820:Silage; D012995:Solubility; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D043366:Cellulose 1,4-beta-Cellobiosidase