Alteration of metal ions improves the activity and thermostability of aminoacylase from hyperthermophilic archaeon Pyrococcus horikoshii.

Motomu Nishioka, Koichi Tanimoto, Noriko Higashi, Harumi Fukada, Kazuhiko Ishikawa, Masahito Taya

Index: Biotechnol. Lett. 30(9) , 1639-43, (2008)

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Abstract

Recombinant L-aminoacylase (PhoACY) from a hyperthermophilic archeon, Pyrococcus horikoshii, is a zinc-containing metalloenzyme. When the zinc was substituted by Mn(2+) or Ni(2+), its specific activity was significantly increased with acetyl-L-methionine as a substrate. The thermostability of PhoACY was improved when it was incubated with 1 mM Zn(2+), Mn(2+) or Ni(2+). The enzyme with external Zn(2+) addition had no significant loss of the activity when held at 90 degrees C for up to 12 h and moreover had more than a 10-fold longer half-life even at 100 degrees C, compared to the enzyme without Zn(2+) addition. A thermostable structure of the enzyme associated with zinc binding is described based on differential scanning calorimetry.