Purification and biochemical characterization of a cellulase from the digestive organs of the short-spined sea urchin Strongylocentrotus intermedius |
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Authors: | Syuto Hasegawa Kazuhiro Ura Hiroyuki Tanaka Takao Ojima Yasuaki Takagi |
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Affiliation: | 2. Laboratory of Comparative Physiology, Graduate School of Fisheries Science, Hokkaido University, 3-1-1 Minato-cho, Hakodate, 041-8611, Japan 1. Graduate School of Fisheries Science, Hokkaido University, 3-1-1 Minato-cho, Hakodate, 041-8611, Japan 3. Laboratory of Marine Molecular Biology, Graduate School of Fisheries Science, Hokkaido University, 3-1-1 Minato-cho, Hakodate, 041-8611, Japan
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Abstract: | We isolated a cellulase from the digestive organs of the short-spined sea urchin Strogylocentrotus intermedius using a combination of ion-exchange chromatography and gel filtration together with an assay for carboxymethylcellulase activity. The isolated cellulase was stained as a single band by Congo red. The molecular weight of the isolated cellulase, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions, was 59?kDa. The isolated cellulase exhibited hydrolytic activity toward carboxymethyl cellulose, with an optimum temperature and pH of 30?°C and pH 8.0, respectively. The thermal stability of the enzyme was characterized by determining the temperature at which activity decreased by 50?% with treatment for 30?min at pH 7.0 and found to be 32?°C. Cellulase activity remained at a high level at 5?C20?°C, which is the growth temperature of the short-spined sea urchin. These results confirm that the short-spined sea urchin should preferably be reared at a water temperature of <20?°C. |
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