title: Purification and characterization of chitinase isozymes from a red algae, Chondrus verrucosus.
abstract: Three seaweed chitinase isozymes (Chi-A, B, and C) were purified from a red algae, Chondrus verrucosus. The molecular weights and isoelectric points were 24.5 kDa and 3.5 for Chi-A, 25.5 kDa and 4.6 for Chi-B, and 24.5 kDa and <3.5 for Chi-C. Optimum pH and temperature were observed at pH 2.0 at 80 degrees C for Chi-A and Chi-C, and at pH 1.0 and 70 degrees C for Chi-B. Toward N-acetylchitooligosaccharide (GlcNAc(n)) (n=2 to 6), Chi-A, B, and C hydrolyzed GlcNAc(5) and GlcNAc(6) and produced GlcNAc(n) (n=2 to 4). GlcNAc(n) (n=3, 4) with the reducing end-side of beta anomer was detected in the hydrolysis products. These results indicate that the reactions of Chi-A, B, and C for GlcNAc(n) were a retaining mechanism similar to that of family 18 chitinase. Toward crystalline chitins, Chi-A, B, and C degraded squid pen beta-chitin more than crab shell or shrimp shell alpha-chitin.
authors.affiliation: Department of Marine Science and Resources, College of Biosource Sciences, Nihon University, Fujisawa, Kanagawa, Japan.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Shirota
authors.forename: Takuya
authors.initials: T
authors.lastname: Sato
authors.forename: Junichi
authors.initials: J
authors.lastname: Sekiguchi
authors.forename: Kouji
authors.initials: K
authors.lastname: Miyauchi
authors.forename: Atsushi
authors.initials: A
authors.lastname: Mochizuki
authors.forename: Masahiro
authors.initials: M
authors.lastname: Matsumiya
mesh_terms: D000595:Amino Acid Sequence; D002688:Chitinases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D044723:Chondrus / Q000201:enzymology*; D004586:Electrophoresis; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012995:Solubility; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D007527:Isoenzymes; D002688:Chitinases