title: Immobilization of Clostridium cellulolyticum D-psicose 3-epimerase on artificial oil bodies.
abstract: The rare sugar D-psicose possesses several fundamental biological functions. D-Psicose 3-epimerase from Clostridium cellulolyticum (CC-DPEase) has considerable potential for use in D-psicose production. In this study, CC-DPEase was fused to the N terminus of oleosin, a unique structural protein of seed oil bodies and was overexpressed in Escherichia coli as a CC-DPEase-oleosin fusion protein. After reconstitution into artificial oil bodies (AOBs), refolding, purification, and immobilization of the active CC-DPEase were simultaneously accomplished. Immobilization of CC-DPEase on AOB increased the optimal temperature but decreased the optimal pH of the enzyme activity. Furthermore, the AOB-immobilized CC-DPEase had a thermal stability and a bioconversion rate similar to those of the free-form enzyme and retained >50% of its initial activity after five cycles of enzyme use. Thus, AOB-immobilized CC-DPEase has potential application in the production of d-psicose at a lower cost than the free-form enzyme.
authors: Institute of Biotechnology, National Taiwan University , Taipei, Taiwan.
authors: Chih-Wen
authors: CW
authors: Tseng
authors: Chien-Yi
authors: CY
authors: Liao
authors: Yuanxia
authors: Y
authors: Sun
authors: Chi-Chung
authors: CC
authors: Peng
authors: Jason T C
authors: JT
authors: Tzen
authors: Rey-Ting
authors: RT
authors: Guo
authors: Je-Ruei
authors: JR
authors: Liu
mesh_terms: D002238:Carbohydrate Epimerases / Q000737:chemistry* / Q000378:metabolism*; D046973:Ruminiclostridium cellulolyticum / Q000201:enzymology*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D005632:Fructose / Q000096:biosynthesis*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D066292:Lipid Droplets / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry*
chemical_list: D004800:Enzymes, Immobilized; D010940:Plant Proteins; C110161:oleosin protein, Sesamum indicum; C003243:psicose; D005632:Fructose; D002238:Carbohydrate Epimerases; C000595678:D-psicose 3-epimerase, Clostridium cellulolyticum