title: Immobilization of Neocallimastix patriciarum xylanase on artificial oil bodies and statistical optimization of enzyme activity.
abstract: A thermally stable and alkalophilic xylanase, XynCDBFV, from Neocallimastix patriciarum was overexpressed in Escherichia coli as a recombinant protein fused to the N-terminus of oleosin, a unique structural protein of seed oil bodies. As a result of the reconstitution of the artificial oil bodies (AOBs), the immobilization of active xylanase was accomplished. Response surface methodology (RSM) was employed for the optimization of the immobilized xylanase activity. The central composite design (CCD) and regression analysis methods were effective for determination of optimized temperature and pH conditions for the AOB-immobilized XynCDBFV. The optimal condition for the highest immobilized xylanase activity (3.93IU/mg of total protein) was observed at 59 degrees C and pH 6.0. Further, AOB-immobilized XynCDBFV retained 50% of its maximal activity after 120min at 60 degrees C, and it could be easily and simply recovered from the surface of the solution by brief centrifugation, and could be reused eight times while retaining more than 60% of its activity. These results proved it is a simple and effective method for direct immobilization of xylanases.
authors.affiliation: Department of Animal Science and Technology, National Taiwan University, Taipei 106, Taiwan.
authors.forename: Ying-Jing
authors.initials: YJ
authors.lastname: Hung
authors.forename: Chi-Chung
authors.initials: CC
authors.lastname: Peng
authors.forename: Jason T C
authors.initials: JT
authors.lastname: Tzen
authors.forename: Ming-Ju
authors.initials: MJ
authors.lastname: Chen
authors.forename: Je-Ruei
authors.initials: JR
authors.lastname: Liu
mesh_terms: D000704:Analysis of Variance; D004591:Electrophoresis, Polyacrylamide Gel; D043364:Endo-1,4-beta Xylanases / Q000032:analysis / Q000378:metabolism*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000032:analysis / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D020088:Neocallimastix / Q000201:enzymology*; D012044:Regression Analysis; D012639:Seeds / Q000378:metabolism*; D013696:Temperature
chemical_list: D004800:Enzymes, Immobilized; D043364:Endo-1,4-beta Xylanases