Purification, properties, and partial amino acid sequence of chitinase from a marine Alteromonas sp. strain O-7.
Chitinase (EC 3.2.1.14) was isolated from the culture supernatant of a marine bacterium, Alteromonas sp. strain O-7. The enzyme (Chi-A) was purified by anion-exchange chromatography (DEAE-Toyopearl 650 M) and gel filtration (Sephadex G-100). The purified enzyme showed a single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The molecular size and pI of Chi-A were 70 kDa and 3.9, respectively. The optimum pH and temperature of Chi-A were 8.0 and 50 degrees C, respectively. Chi-A was stable in the range of pH 5-10 up to 40 degrees C. Among the main cations, such as Na+, K+, Mg2+, and Ca2+, contained in seawater, Mg2+ stimulated Chi-A activity. N-Bromosuccinimide and 2-hydroxy-5-nitrobenzyl bromide inhibited Chi-A activity. The amino-terminal 27 amino acid residues of Chi-A were sequenced. This enzyme showed sequence homology with chitinases from terrestrial bacteria such as Serratia marcescens QMB1466 and Bacillus circulans WL-12.
Tsujibo
H
H
Osaka University of Pharmaceutical Sciences, Japan.
Yoshida
Y
Y
Miyamoto
K
K
Imada
C
C
Okami
Y
Y
Inamori
Y
Y
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002686:Chitin / Q000378:metabolism; D002688:Chitinases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006088:Gram-Negative Aerobic Bacteria / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008969:Molecular Sequence Data; D012492:Salts / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid; D013696:Temperature
D000596:Amino Acids; D012492:Salts; D002686:Chitin; D002688:Chitinases