Use of enzymatic cell wall degradation for improvement of protein extraction from Chondrus crispus,Gracilaria verrucosa and Palmaria palmata |
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Authors: | Joël Fleurence Laurence Massiani Odile Guyader Serge Mabeau |
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Institution: | (1) Centre d'Etude et de Valorisation des Algues, BP 3, 22610 Pleubian, France;(2) Present address: Laboratoire Qualité et Physico-Chimie, IFREMER, BP 1105, 44311 Nantes, France |
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Abstract: | The effect of polysaccharidases (κ-carrageenase, β-agarase, xylanase, cellulase) on the protein extraction from three rhodophytes
has been studied. The kinetic parameters (apparent V
m, apparent K
m) and the optimum activity conditions (pH, temperature) of each enzyme were determined by using pure substrates. All the tested
enzymes possess Michaelis Menten mechanism with estimated substrate saturating concentrations of 8 000 mg l−1(carrageenan) for κ-carrageenase, 8 000 mg l−1 (agar) for β-agarase, 5000 mg l−1 (xylane) for β-xylanase and 6 000 mg l−1 (carboxymethylcellulose) for cellulase. The optimum activity conditions are pH 6.5–6.8 at 45°C for carrageenase, pH 6–6.5
at 55°C for agarase, pH 5 at 55°C for xylanase and pH 3.8 at 50°C for cellulose. Different alga/enzymes couples (κ-carrageenase/Chondrus crispus, β-agarase/Gracilaria verrucosa, β-xylanase/Palmaria palmata) were tested under the optimum activity conditions. Alga/cellulase + specific enzyme (e.g. Chondrus crispus/carrageenase + cellulase) systems were also studied at the optimum activity conditions of a specific enzyme (e.g. carageenase).
The use of the only cellulose was also tested on each alga. Except for Palmaria palmata, the highest protein yields were observed with the procedures using cellulase coupled with carrageenase or agarase for an
incubation period limited to 2 h. The Chondrus crispus/carrageenase + cellulose and Gracilaria verrucosa/agarase + cellulase systems gave ten-fold and three-fold improvements, respectively, in protein extraction yield as compared
to the enzyme-free blank procedure. The combined action of xylanase and cellulose on protein extraction from Palmaria palmata does not significantly improve protein yield. The best overall protein yield for P. palmata is for P. palmata/xylanase with a 14-h incubation time. This study shows the interest in the use of a polysaccharidase mixture for improving
protein extractibility from certain rhodophytes. This biotechnology approach, adapted from procedures for protoplast production
or enzymatic liquefaction of higher plants, could be tested as an alternative method to obtain proteins from seaweeds of nutritional
interest. |
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Keywords: | red seaweeds polysaccharidases improvement of protein extraction |
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