title: pH-dependent stability of EGX, a multi-functional cellulase from mollusca, Ampullaria crossean.
abstract: The cellulase activity and stability of EGX, a multi-functional cellulase previously purified from the mollusca Ampullaria crossean, was systematically studied under different pH. The pH induced con-formation and stability change of EGX have been investigated by using the intrinsic fluorescence, ANS fluorescence and CD spectrum. It has been found that the conformation and activity of this cellulase were strongly dependent on the pH. EGX was stable for both the enzyme activity and the conformation from pH 5.6 to pH 7.4. As shown by intrinsic and ANS fluorescence, no red shift of emission maximum occurred and a negligible intensity change was observed at pH 5.6-7.4. The activity of EGX remained about 80% in pH 5.6-7.4 and obviously decreased out of side the pH range. Urea-induced changes in EGX at pH 5.4 and pH 8.0 were measured by intrinsic fluorescence and CD spectrum. At pH 5.4, a significantly red shift of emission maximum occurred when the concentration of urea was 5 M compared to the concentration was 3 M at pH 8.0. The alpha-helix at pH 5.4 was 40.51% in the absence of urea and 31.04% in the presence of 4 M urea. At pH 8.0 the alpha-helix was 7.23% in the presence of 4 M urea. The data indicated that EGX was much susceptible to urea-induced unfolding at pH 8.0 and much stable at pH 5.4. The greater pH dependent stability of EGX may allow the enzyme to adequately catalyze the hydrolysis of cellulosic materials under natural or industrial extreme conditions.
authors.affiliation: Key Laboratory of Proteomics, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, Graduate School of the Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Wen-Ying
authors.initials: WY
authors.lastname: Li
authors.forename: Ji
authors.initials: J
authors.lastname: Wang
authors.forename: Yan-Hong
authors.initials: YH
authors.lastname: Li
authors.forename: Ming
authors.initials: M
authors.lastname: Ding
authors.forename: Gen-Jun
authors.initials: GJ
authors.lastname: Xu
authors.forename: Lan-Ying
authors.initials: LY
authors.lastname: Liu
authors.forename: Fu-Kun
authors.initials: FK
authors.lastname: Zhao
mesh_terms: D000818:Animals; D044602:Cellulases / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D002942:Circular Dichroism; D004795:Enzyme Stability; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008974:Mollusca / Q000201:enzymology*; D011487:Protein Conformation / Q000187:drug effects; D011489:Protein Denaturation / Q000187:drug effects; D017433:Protein Structure, Secondary; D013050:Spectrometry, Fluorescence; D014508:Urea / Q000494:pharmacology
chemical_list: D014508:Urea; D044602:Cellulases; C493084:EGX, Ampullaria crossean