title: A useful method integrating production and immobilization of recombinant cellulase.
abstract: The development of cellulase-based bioprocess is afflicted by the processing efficiency of enzymes. To address this issue, a method based on artificial oil bodies (AOBs) was proposed to integrate production and immobilization of recombinant cellulase. First, the heterologous endoglucanase (celA), cellobiohydrolase (celK), and β-glucosidase (gls) genes were individually fused with oleosin, a structural protein of plant seed oils. After expression in Escherichia coli, each fusion protein of insolubility was mixed together with plant oils. AOBs were assembled by subjecting the mixture to sonication. Consequently, active CelA, CelK, and Gls were resumed and co-immobilized on AOBs surface. Finally, the assembly condition (including the protein ratio) and the reaction condition were further optimized by response surface methodology. The resulting AOBs-bound cellulase remained stable for 4 cycles of cellulose-hydrolyzed reactions. Overall, the result shows a promise of this proposed approach for processing recombinant cellulase, which may provide a facile method to investigate optimum combination of cellulase components towards various cellulosic materials.
authors.affiliation: Department of Medical Laboratory Science and Biotechnology, China Medical University, No. 91, Hsueh-Shih Road, Taichung, 40402, Taiwan.
authors.forename: Chung-Jen
authors.initials: CJ
authors.lastname: Chiang
authors.forename: Po Ting
authors.initials: PT
authors.lastname: Chen
authors.forename: Chien Yu
authors.initials: CY
authors.lastname: Yeh
authors.forename: Zei Wen
authors.initials: ZW
authors.lastname: Wang
authors.forename: Yun-Peng
authors.initials: YP
authors.lastname: Chao
mesh_terms: D001419:Bacteria / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001671:Biochemistry / Q000379:methods*; D044602:Cellulases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D048013:Acetivibrio thermocellus / Q000737:chemistry / Q000201:enzymology*; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010938:Plant Oils / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D004800:Enzymes, Immobilized; D010938:Plant Oils; D010940:Plant Proteins; D011994:Recombinant Proteins; D044602:Cellulases