Protoplast production in Chondrus crispus gametophytes (gigartinales, rhodophyta)

Gall, Y. Le; Braud, Jean-Paul; Kloareg, Bernard

doi:10.1007/bf00270058

Cited by 48 publications

(19 citation statements)

References 11 publications

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“…The efficiency of procedure using simultaneously carrageenase and cellulase on Chondrus crispus suggest that the destruction of cellulose facilitates access to carrageenan. This fact indirectly confirms the results of Le Gall et al (1990), who found that the carrageenase-cellulase couple gives optimal production of protoplasts from Chondrus crispus. An increase of incubation time does not appear to be an important factor to improve the protein extraction from Chondrus crispus (Table 4).…”

Section: Discussionsupporting

confidence: 91%

“…Epiphytes and other contaminants (sand, shells) were eliminated by successive washings with sea water and deionised water. The algal material (10 g) was cut into 0.5-1 cm pieces (Le Gall et al, 1990), suspended in the incubation medium containing appropriate enzymes and buffers (Tables 1, 2), and incubated for 2 or 14 h under the optimum activity conditions (pH, temperature) for the enzyme tested (Table 1). NaCI (500 mM), MgCI2 (40 mM), KC1 (5 mM) were added into the incubation medium as described by Le Gall et al (1991) for the production of Chondrus crispus protoplasts with carrageenase and cellulase.…”

Section: Methodsmentioning

confidence: 99%

“…However, the presence of cell wall polysaccharides limits the efficiency of classical protein extraction procedures (Jordan & Vilter, 1991). The degradation of cell wall polysaccharides by enzymes is a procedure frequently used for protoplast production (Le Gall et al, 1990;Liu et aL, 1992;Cachot et al, 1994) and has been used for the isolation of extensin, a protein linked to cell wall polysaccharides of higher plants (Lamport, 1969). A study performed on the red alga Porphyra yezoensis showed that the protoplasts production using hydrolitic enzymes improves accessibility to the proteic fraction (Amano & Noda, 1990).…”

Section: Introductionmentioning

confidence: 99%

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Use of enzymatic cell wall degradation for improvement of protein extraction from Chondrus crispus, Gracilaria verrucosa and Palmaria palmata

et al. 1995

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The effect of polysaccharidases (;-carrageenase, P-agarase, xylanase, cellulase) on the protein extraction from three rhodophytes has been studied. The kinetic parameters (apparent V,, apparent Kin) 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 1-(carrageenan) for -carrageenase, 8000 mg 1-' (agar) for B-agarase, 5000 mg 1 -' (xylane) for -xylanase and 6000 mg 1 -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 0 C for agarase, pH 5 at 55°C for xylanase and pH 3.8 at 50°C for cellulase. Different alga/enzymes couples (-carrageenase/Chondrus crispus, /3-agaraselGracilaria verrucosa, /-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 cellulase 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 + cellulase 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 cellulase 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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Section: Discussionsupporting

confidence: 91%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Use of enzymatic cell wall degradation for improvement of protein extraction from Chondrus crispus, Gracilaria verrucosa and Palmaria palmata

et al. 1995

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“…The addition volume was adapted to the new fresh weight in such a way that the nutrient load for each treatment was kept constant during the experiment. During the last 2 wk of the experiment, 47.8 μM GeO 2 was added to the medium to suppress diatom growth (Le Gall et al 1990). The experiment lasted 32 d, after which all units were collected and weighed; the stolon length produced was recorded and the final number of assimilators was counted.…”

Section: Methodsmentioning

confidence: 99%

Nitrogen load and irradiance affect morphology, photosynthesis and growth of Caulerpa prolifera (Bryopsidales: Chlorophyta)

Malta¹,

Ferreira²,

Vergara³

et al. 2005

Mar. Ecol. Prog. Ser.

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The effect of nitrogen (N) load and irradiance on morphology, growth and photosynthetic performance was studied in the green macroalga Caulerpa prolifera (Forsskål) J. V. Lamouroux from the Gulf of Cádiz (south Spain). Constant growth rates were obtained for thalli growing at different N loads, which could be fitted to tissue N using the Droop equation, rendering a maximum growth rate of 0.09 d -1 , a minimum tissue N level of 1.71% dry weight (DW) and a critical tissue N of 5.2% DW. N limitation had no effects on F v /F m (maximum quantum yield of chlorophyll a fluorescence). Stolon production was significantly highest at low N loads; a reverse trend was observed for assimilator production. In a second experiment, algae were subjected to combinations of high and low N loads (HN and LN) and irradiance (HL and LL) levels. Highest growth rates were observed in the HNLL treatment, whereas the reverse combination rendered the lowest growth rate. High irradiance and high N load both led to increased biomass allocation to assimilators; at low N, the bulk of the biomass (> 75% in the HLLN treatment) was allocated to the stolons. HN had a positive effect on F v /F m , and HL had a negative effect. HL algae had a higher capacity for non-photochemical quenching. Despite its prolific nature, C. prolifera should be characterised as a slow-growing, but highly nitrophilic alga which has the capacity to forage for nutrients by allocating biomass to the stolons. KEY WORDS: Caulerpa prolifera · Clonal growth · Morphology · Nitrogen foraging · Growth rate · PhotosynthesisResale or republication not permitted without written consent of the publisher

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“…The biotechnology of enzymes and enzyme degraded products of carrageenan is still in infancy compared to that of agar, alginate, starch [3] or pectin [4]. Carrageenases which hydrolyse 1,4 linkages in carrageenan to a series of homologous, even numbered oligosaccharides [5] are useful tool for the structural analysis of the cell walls and protoplast isolation from red algae [6,7]. The sulfated carrageeno-oligosaccharides have also drawn considerable interest [8] owing to their diverse biological and physiological activities including anticoagulation [9], anti-inflammation [10], anti-thrombosis [11], antitumor activity [12] and viral inactivation [13], which depend on structural parameters such as carbohydrate structure, molecular mass, degree of sulfate esterification, and the linking position of sulpho groups [14].…”

Section: Introductionmentioning

confidence: 99%

Increased Production of Carrageenase by <i>Pseudomonas aeruginosa</i> ZSL-2 Using Taguchi Experimental Design

Ziayoddin

Lalitha

Shinde

2014

ILNS

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The culture conditions for the production of carrageenase were optimized using one-factor-at-atime method combined with orthogonal array design. With the one-factor-at-a-time method revealed optimal conditions for carrageenase production were 24 h of fermentation period, 28 °C incubation temperature at pH 8.0 with NaNO 3 as nitrogen source and carrageenan as carbon source in MMS media. Further optimization of carrgeenase production by using orthogonal experimental design L 9 (3 4 ) with four factors, temperature, pH, NH 4 NO 3 and carrageenan with their relevant levels revealed optimised conditions for carrageenase production were temperature of 28 °C, pH 8.0, 2 g L -1 NaNO 3 and 2 g L -1 carrageenan. The order of the factors affecting the fermentation process was found to be temperature > pH > NaNO 3 > carrageenan. The temperature played a significant role on the carrageenase production. Higher carrageenase yield with activity of 0.542 ±0.045 U ml -1 was obtained in the optimised medium when compared to those of basal medium. Carrageenase hydrolysed products of carrageenan were identified by LC-ESI-MS as neocarrabiose, neocarrabiose-4 sulfate, neocarratetraose, neocarratetraose-4 sulfate, anhydrogalactose, galactose, galactose-4 sulphate and sulphate.

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Protoplast production in Chondrus crispus gametophytes (gigartinales, rhodophyta)

Cited by 48 publications

References 11 publications

Use of enzymatic cell wall degradation for improvement of protein extraction from Chondrus crispus, Gracilaria verrucosa and Palmaria palmata

Use of enzymatic cell wall degradation for improvement of protein extraction from Chondrus crispus, Gracilaria verrucosa and Palmaria palmata

Nitrogen load and irradiance affect morphology, photosynthesis and growth of Caulerpa prolifera (Bryopsidales: Chlorophyta)

Increased Production of Carrageenase by <i>Pseudomonas aeruginosa</i> ZSL-2 Using Taguchi Experimental Design

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