Analysis of the Hyperthermophilic Chitinase from Pyrococcus furiosus : Activity toward Crystalline Chitin
Chitinase [EC 220.127.116.11] is an enzyme that can hydrolyze the β-1,4 linkage between <I>N</I>-acetyl-<small>D</small>-glucosamine in chitin. In the genome database of the hyperthermophilic archaeon <I>Pyrococcus furiosus</I>, we found two adjacent genes (PF1233 and PF1234) homologous to those of the chitinase of <I>Thermococcus kodakaraensis</I>. In the cultured medium of <I>P. furiosus</I>, however, no chitinase activity was detected. On analysis of the structural gene of <I>P. furiosus</I>, it appears that one nucleotide insertion in PF1234 caused a frame shift and separated a gene. By deletion of one nucleotide in PF1234, the best match was achieved between chitinases of <I>T. kodakaraenesis</I> and <I>P. furiosus</I>. We succeeded in constructing an artificial recombinant chitinase exhibiting hydrolytic activity toward not only colloidal but also crystalline chitins at high temperature. Furthermore, by analyzing the characteristics of the domains, a recombinant enzyme comprising two domains exhibiting high activity toward crystalline chitin was prepared.
- Bioscience, biotechnology, and biochemistry
Bioscience, biotechnology, and biochemistry 70(7), 1696-1701, 2006-07-23
Japan Society for Bioscience, Biotechnology, and Agrochemistry