title: Preparation and characterization of non-covalently immobilized amylosucrase using a pH-dependent autoprecipitating carrier.
abstract: In an effort to use a biocatalyst repeatedly with acceptable stability, amylosucrase from Neisseria polysaccharea (NpAS) was non-covalently immobilized on a pH-dependent autoprecipitating polymer, Eudragit L100. About 87% of the enzyme activity and 96% of the protein were recovered after the immobilization process. The immobilized NpAS showed significantly improved thermostability, whereas no difference was observed in the temperature and pH profiles of activity between the native and immobilized NpAS. To evaluate the reusability of the immobilized NpAS, repeated production of linear α-(1,4)-glucans was carried out at 35°C with 0.1M sucrose as substrate. Although only 71-74% of the protein was recovered after each reaction cycle, high stability index was observed from 0.93 to 0.96. Reusability of the immobilized NpAS was further confirmed by comparing its linear α-(1,4)-glucan products with that produced by native one. Almost identical production yield and molecular size distribution of the linear α-(1,4)-glucans were obtained between them.
authors.affiliation: Department of Food Science & Technology, BK21 Project Team, and Carbohydrate Bioproduct Research Center, Sejong University, 98 Gunja-Dong, Gwangjin-Gu, Seoul 143-747, Republic of Korea.
authors.forename: Ren
authors.initials: R
authors.lastname: Wang
authors.forename: Jung-Hwan
authors.initials: JH
authors.lastname: Kim
authors.forename: Bum-Soo
authors.initials: BS
authors.lastname: Kim
authors.forename: Cheon-Seok
authors.initials: CS
authors.lastname: Park
authors.forename: Sang-Ho
authors.initials: SH
authors.lastname: Yoo
mesh_terms: D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005964:Glucosyltransferases / Q000378:metabolism*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D009343:Neisseria / Q000201:enzymology
chemical_list: D004800:Enzymes, Immobilized; D005964:Glucosyltransferases; C019700:amylosucrase