Characterization of a cellulase from the haemolymph of the giant African snail (Archachatina marginata)

Abstract

The giant African snails (Archachatina marginata) are herbivorous tropical gastropods. They are commonly located in areas littered with decaying vegetable matters which they feed on. When the haemolyph of this organism was passed through Sephadex G-200, the resin was digested. The ability of snails to digest vegetable matters and the digestion of Sephadex G-200 resin by the haemolyph led to the suspicion of the presence of cellulase in snail haemolyph. Cellulase from haemolyph of the giant African snail was purified to homogeneity by a combination of gel filtration on BioGel P-300 and ionexchange chromatography on DEAE-Sephadex. The homogeneity of the pure enzyme was adjudged by polyacrylamide gel electrophoresis in the absence and presence of sodium dodecyl sulphate. The subunit molecular weight was 16,875±1,556 daltons and the apparent molecular weight as determined by gel filtration was 52,000 daltons. Thus, the enzyme is at least a dimer. The enzyme had a specific activity of 1359.09 units/mg of protein. The Michaelis-Menten constant (K m) for carboxymethyl (CM)cellulose was 5.17± 0.74 mg/ml and the maximum velocity (V max) was 1067±195 units/ml. The enzyme did not degrade salicin, cellobiose and o-nitrophenyl-β-D-glucopyranoside. In addition, its activities towards filter paper and cotton wool were insignificant compared with its activity towards CM-Cellulose. Linamarin was found to inhibit the action of the enzyme to about 18%. The high specificity for CMcellulose and the rapid decrease in the viscosity of CM-cellulose suggests that the enzyme is likely an endoglucanase.

Isolasi dan Karakterisasi Enzim Selulase dari Keong Sawah Pila ampullaceal Menggunakan Substrat Serbuk Gergaji Kayu

“…Enzim selulase dapat disolasi dari mikroorganisme, hewan, tumbuhan. Telah banyak dilaporkan bahwa selulase dapat diisolasi dari hewan tingkat rendah seperti moluska, antara lain keong Mudalia dilatata [10], (Farrisdkk., 1994), keong mas Pomacea canaliculata [11], bekicot Achatina fulica [12,13], siput air tawar Menggunakan Substrat Serbuk Gergaji Kayu Ampullaria crossean [14], dan keong afrika Archatina marginata [15].…”