An endo-beta-1,4-mannanase, AkMan, from the common sea hare Aplysia kurodai.
A mannan-degrading enzyme was isolated from the digestive fluid of the common sea hare Aplysia kurodai by ammonium sulfate fractionation followed by conventional column chromatography. The purified enzyme, named AkMan in the present paper, showed a single band with an approximate molecular mass of 40,000 Da on SDS-PAGE and preferably degraded a linear beta-1,4-mannan from green algae Codium fragile producing tri- and disaccharides. The optimal temperature of AkMan was 55 degrees C at pH 7.0 and temperature that caused 50% inactivation of AkMan during a 20-min incubation was 52 degrees C. AkMan retained high activity at pH 4.0-7.5 and was not inactivated in such acidic pH range by the incubation at 40 degrees C for 20 min. AkMan could degrade glucomannan from konjak root and galactomannan (tara gum and guar gum) as well as the linear beta-1,4-mannan, while not carboxymethyl cellulose, agarose, dextran and xylan. These results indicate that AkMan is a typical endo-beta-1,4-mannanase (EC 3.2.1.78) splitting internal beta-1,4-mannosyl linkages of mannan. The N-terminal and internal amino-acid sequences of AkMan shared approximately 55% amino-acid identity to the corresponding sequences of an abalone beta-1,4-mannanase, HdMan, which belongs to glycosyl hydrolase family 5 (GHF5). Thus, AkMan was also regarded as a member of GHF5 beta-1,4-mannanases.
Zahura
Umme Afsari
UA
Laboratory of Marine Biotechnology and Microbiology, Graduate School of Fisheries Sciences, Hokkaido University, Hakodate, Hokkaido, 041-8611, Japan.
Rahman
Mohammad Matiur
MM
Inoue
Akira
A
Tanaka
Hiroyuki
H
Ojima
Takao
T
D000595:Amino Acid Sequence; D000818:Animals; D001048:Aplysia / Q000201:enzymology*; D002240:Carbohydrate Sequence; D004064:Digestive System / Q000201:enzymology; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents / Q000494:pharmacology; D008351:Mannans / Q000737:chemistry / Q000378:metabolism; D008361:Mannosidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D008670:Metals / Q000494:pharmacology; D008969:Molecular Sequence Data; D013379:Substrate Specificity; D013696:Temperature
D007202:Indicators and Reagents; D008351:Mannans; D008670:Metals; D008361:Mannosidases; C041867:endo-1,4-beta-D-mannanase