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Development of a novel immobilization method for enzymes from hyperthermophiles |
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| Authors: | Kosuke Minamihata Masamichi Tokunaga Noriho Kamiya Shiro Kiyoyama Haruhiko Sakuraba Toshihisa Ohshima Masahiro Goto |
| Institution: | (1) Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, 744 Motooka, Fukuoka 819-0395, Japan;(2) Center for Future Chemistry, Kyushu University, 744 Motooka, Fukuoka 819-0395, Japan;(3) Department of Chemical Science and Engineering, Miyakonojo National College of Technology, Miyakonojo Miyazaki, 885-8567, Japan;(4) Department of Applied Biological Science, Faculty of Agriculture, Kagawa University, Kagawa 761-0795, Japan;(5) Microbial Genetic Division, Institute of Genetic Resources, Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan; |
| Abstract: | Peptide tags containing tyrosines (Y-tag) were introduced at the C-terminus of a hyperthermophilic enzyme, alkaline phosphatase from Pyrococcus furiosus (PfuAP). Immobilization of the recombinant PfuAPs onto water-in-oil-in-water (W/O/W) type microcapsules was performed by an in situ polymerization method. All the recombinant PfuAPs prepared in this study were quantitatively immobilized onto microcapsules. The PfuAP-immobilized microcapsules showed no significant loss of enzymatic activity until the 5th round of assays. This result implies that the recombinant PfuAPs were covalently immobilized onto microcapsules. Immobilized PfuAP tagged with a Y-tag having the sequence GGYYY exhibited approximately a twofold higher catalytic activity compared with the wild-type PfuAP. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. |
| Keywords: | Enzyme Hyperthermophilic enzyme Immobilization Microcapsules Polymerization Tyrosine Pyrococcus |
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