Cloning and Sequencing of a Chitinase Gene from Vibrio alginolyticus H-8

概要

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A gene from Vibrio alginolyticus H-8, encoding chitinase, designated as chitinase B, was cloned by the shot-gun method using pUC118 and sequenced. The open reading frame consisted of 846 amino acids including a signal peptide. The molecular mass of the enzyme estimated based on the amino acid sequence data was 90 kDa. The N-terminal amino acid sequence of the enzyme was different from that of chitinase C1 which we had previously reported. This cloned chitinase B was considered one out of four chitinases(A, B, D, and E) which had been newly isolated from the culture broth and cell extract of V.alginolyticus H-8. The gene contained a chitin-binding domain and typical conserved regions of chitinases reported previously. The deduced amino acid sequence of the cloned chitinase B showed high sequence homology with the chitinase from V.parahaemolyticus(84% identity) and the chitinase from V.anguillarum(76.6%), but low sequence homology with the chitinase from V.harveyi(24.4%), and the chitodextrinase from V.furnissii(23.9%). Chitinase E found in cell extract is considered an intracellular chitinase which is different from chitodextrinases.
社団法人日本生物工学会の論文
2000-05-25