Development of a novel immobilization method for enzymes from hyperthermophiles

Kosuke Minamihata, Masamichi Tokunaga, Noriho Kamiya, Shiro Kiyoyama, Haruhiko Sakuraba, Toshihisa Ohshima, Masahiro Goto

Research output: Contribution to journalArticle

3 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1037-1041
Number of pages5
JournalBiotechnology letters
Volume31
Issue number7
DOIs
Publication statusPublished - Jul 1 2009

Fingerprint

Immobilization
Capsules
Enzymes
Phosphatases
Peptides
Water
Assays
Catalyst activity
Pyrococcus furiosus
Polymerization
Alkaline Phosphatase
Tyrosine
Oils

All Science Journal Classification (ASJC) codes

  • Biotechnology

Cite this

Development of a novel immobilization method for enzymes from hyperthermophiles. / Minamihata, Kosuke; Tokunaga, Masamichi; Kamiya, Noriho; Kiyoyama, Shiro; Sakuraba, Haruhiko; Ohshima, Toshihisa; Goto, Masahiro.

In: Biotechnology letters, Vol. 31, No. 7, 01.07.2009, p. 1037-1041.

Research output: Contribution to journalArticle

Minamihata, K, Tokunaga, M, Kamiya, N, Kiyoyama, S, Sakuraba, H, Ohshima, T & Goto, M 2009, 'Development of a novel immobilization method for enzymes from hyperthermophiles', Biotechnology letters, vol. 31, no. 7, pp. 1037-1041. https://doi.org/10.1007/s10529-009-9974-8
Minamihata, Kosuke ; Tokunaga, Masamichi ; Kamiya, Noriho ; Kiyoyama, Shiro ; Sakuraba, Haruhiko ; Ohshima, Toshihisa ; Goto, Masahiro. / Development of a novel immobilization method for enzymes from hyperthermophiles. In: Biotechnology letters. 2009 ; Vol. 31, No. 7. pp. 1037-1041.
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