Production of ethanol from mannitol by Zymobacter palmae

Extracts from brown seaweeds could possibly be fermented to ethanol, particularly seaweeds harvested in the autumn, which contain high levels of easily extractable laminaran and mannitol. Few microorganisms are able to utilise mannitol as a substrate for ethanol production and Zymobacter palmae was tested for this purpose. Bacterial growth as well as ethanol yield depended on the amount of oxygen present. Strictly anaerobic growth on mannitol was not observed. At excessive aeration, a change in the fermentation pattern was observed with high production of acetate and propionate. Under oxygen-limiting conditions, the bacteria grew and produced ethanol in a synthetic mannitol medium with a yield of 0.38 g ethanol (g mannitol)⁻¹. Z. palmae was also successfully applied for fermentation of mannitol from Laminaria hyperborea extracts. Journal of Industrial Microbiology & Biotechnology (2000) 24, 51–57. Received 27 June 1999/ Accepted in revised form 23 September 1999     

Fermentation study on Saccharina latissima for bioethanol production considering variable pre-treatments

Climate change, fuel security and economics are creating a requirement for alternative, renewable fuels. Bioethanol is currently produced from land-based crops but in future marine biomass could be used as an alternative biomass source for bioethanol production. Macroalgae such as Laminaria spp. grow in abundance around the United Kingdom, reaching >4 m in length and containing up to 55% dry weight of the carbohydrates laminarin and mannitol. Using enzymes, these can be hydrolysed and converted to glucose and fructose, which in turn can be utilised by yeasts to produce ethanol. In previous studies on ethanol production from macroalgae, pre-treatment was at 65°C, pH 2 for 1 h prior to fermentation. This paper shows that these pre-treatments are not required for the fermentations conducted, with higher ethanol yields being achieved in untreated fermentations than in those with altered pH or temperature pre-treatments. This result was seen in fresh and defrosted macroalgae samples using Saccharomyces cerevisiae and 1 U kg⁻¹ laminarinase.     

Ethanol production from seaweed extract

Extracts from Laminaria hyperborea could possibly be fermented to ethanol commercially. In particular, seaweed harvested in the autumn contains high levels of easily extractable laminaran and mannitol. Four microorganisms were tested to carry out this fermentation, one bacterium and three yeasts. Only Pichia angophorae was able to utilise both laminaran and mannitol for ethanol production, and its substrate preferences were investigated in batch and continuous cultures. Laminaran and mannitol were consumed simultaneously, but with different relative rates. In batch fermentations, mannitol was the preferred substrate. Its share of the total laminaran and mannitol consumption rate increased with oxygen transfer rate (OTR) and pH. In continuous fermentations, laminaran was the preferred substrate at low OTR, whereas at higher OTR, laminaran and mannitol were consumed at similar rates. Optimisation of ethanol yield required a low OTR, and the best yield of 0.43 g ethanol (g substrate)⁻¹ was achieved in batch culture at pH 4.5 and 5.8 mmol O₂ l⁻¹ h⁻¹. However, industrial production of ethanol from seaweed would require an optimisation of the extraction process to yield a higher ethanol concentration. Journal of Industrial Microbiology & Biotechnology (2000) 25, 249–254. Received 25 February 2000/ Accepted in revised form 05 August 2000     

OSMOTIC ADJUSTMENT IN LAMINARIA DIGITATA (PHAEOPHYTA) WITH PARTICULAR REFERENCE TO SEASONAL CHANGES IN INTERNAL SOLUTE CONCENTRATIONS1

Laboratory studies have indicated that Na⁺, K⁺ (together with Cl^? the presumed counter-ion to these cations), NO₃^? and mannitol represent the major cellular osmotica in Laminaria digitata (Huds.) Lamour. The cellular content of NO₃^? (together with a fraction of the K⁺ pool which acts as the counter-ion to NO₃^?) was found to be inversely proportional to that of mannitol, suggesting that L. digitata maintains a constant turgor by means of an isotonic substitution between these compounds. An analysis of the seasonal changes in solute content in an Arbroath (Scotland) population of L. digitata confirmed this hypothesis and indicated that the total pool of stored photosynthate was partitioned between the interconvertible carbohydrates mannitol and laminaran (which has a much lower osmotic potential than mannitol) depending on the size of the cellular pool of NO₃^?.     

Carbohydrate degradation and methane production during fermentation ofLaminaria saccharina (Laminariales,Phaeophyceae)

Anaerobic digestion of the brown algaLaminaria saccharina (L.) Lamour. harvested in spring and autumn was carried out at controlled laboratory conditions in stirred fermentor systems. Due to the normal seasonal variations, the autumn material had a much higher content of carbohydrates such as mannitol and laminaran. Both batch and semi-continuous feeding conditions were investigated for periods up to 800 h, with inoculum provided from previous kelp fermentations. In batch cultures, the methane yield from the autumn material was doubled compared to that of the spring material. Semi-continuous conditions gave more similar methane yields for both raw materials, 0.22 and 0.27 l CH₄ per g VS for spring and autumn material, respectively. In all experiments, mannitol and laminaran were reduced to less than 5 of the initial values within 24–48 hours after inoculation, whereas 30 of the alginate content was detectable even after 30 days. Viscometry revealed that this material was severely depolymerized, and alginate lyase activity was found to develop rapidly in all cultures. Although mannitol and laminaran were fermented much faster than alginate, the total accumulated methane yields seemed to be determined by the total carbohydrate content of the raw material during extended semi-continuous feeding.(*author for correspondence)     

Construction of bioengineered yeast platform for direct bioethanol production from alginate and mannitol

Brown macroalgae are a sustainable and promising source for bioethanol production because they are abundant in ocean ecosystems and contain negligible quantities of lignin. Brown macroalgae contain cellulose, hemicellulose, mannitol, laminarin, and alginate as major carbohydrates. Among these carbohydrates, brown macroalgae are characterized by high levels of alginate and mannitol. The direct bioconversion of alginate and mannitol into ethanol requires extensive bioengineering of assimilation processes in the standard industrial microbe Saccharomyces cerevisiae. Here, we constructed an alginate-assimilating S. cerevisiae recombinant strain by genome integration and overexpression of the genes encoding endo- and exo-type alginate lyases, DEH (4-deoxy-l-erythro-5-hexoseulose uronic acid) transporter, and components of the DEH metabolic pathway. Furthermore, the mannitol-metabolizing capacity of S. cerevisiae was enhanced by prolonged culture in a medium containing mannitol as the sole carbon source. When the constructed strain AM1 was anaerobically cultivated in a fermentation medium containing 6% (w/v) total sugars (approximately 1:2 ratio of alginate/mannitol), it directly produced ethanol from alginate and mannitol, giving 8.8 g/L ethanol and yields of up to 32% of the maximum theoretical yield from consumed sugars. These results indicate that all major carbohydrates of brown macroalgae can be directly converted into bioethanol by S. cerevisiae. This strain and system could provide a platform for the complete utilization of brown macroalgae.     

Iodine contributes to osmotic acclimatisation in the kelp Laminaria digitata (Phaeophyceae)

Iodide (I^?) retained by the brown macroalga Laminaria digitata at millimolar levels, possesses antioxidant activities, but the wider physiological significance of its accumulation remains poorly understood. In its natural habitat in the lower intertidal, L. digitata experiences salinity changes and osmotic homeostasis is achieved by regulating the organic osmolyte mannitol. However, I^? may also holds an osmotic function. Here, impacts of hypo- and hypersaline conditions on I^? release from, and accumulation by, L. digitata were assessed. Additionally, mannitol accumulation was determined at high salinities, and physiological responses to externally elevated iodine concentrations and salinities were characterised by chl a fluorometry. Net I^? release rates increased with decreasing salinity. I^? was accumulated at normal (35 S _A) and high salinities (50 S _A); this coincided with enhanced rETR_max and q_P causing pronounced photoprotection capabilities via NPQ. At 50 S _A elevated tissue iodine levels impeded the well-established response of mannitol accumulation and prevented photoinhibition. Contrarily, low tissue iodine levels limited photoprotection capabilities and resulted in photoinhibition at 50 S _A, even though mannitol was accumulated. The results indicate a, so far, undescribed osmotic function of I^? in L. digitata and, thus, multifunctional principles of this halogen in kelps. The osmotic function of mannitol may have been substituted by that of I^? under hypersaline conditions, suggesting a complementary role of inorganic and organic solutes under salinity stress. This study also provides first evidence that iodine accumulation in L. digitata positively affects photo-physiology.     

Quantifying Actual and Theoretical Ethanol Yields for Switchgrass Strains Using NIRS Analyses

Quantifying actual and theoretical ethanol yields from biomass conversion processes such as simultanteous saccharification and fermentation (SSF) requires expensive, complex fermentation assays, and extensive compositional analyses of the biomass sample. Near-infrared reflectance spectroscopy (NIRS) is a non-destructive technology that can be used to obtain rapid, low-cost, high-throughput, and accurate estimates of agricultural product composition. In this study, broad-based NIRS calibrations were developed for switchgrass biomass that can be used to estimate over 20 components including cell wall and soluble sugars and also ethanol production and pentose sugars released as measured using a laboratory SSF procedure. With this information, an additional 13 complex feedstock traits can be determined including theoretical and actual ethanol yields from hexose fermentation. The NIRS calibrations were used to estimate feedstock composition and conversion information for biomass samples from a multi-year switchgrass (Panicum virgatum L.) biomass cultivar evaluation trial. There were significant differences among switchgrass strains for all biomass conversion and composition traits including actual ethanol yields, ETOHL (L Mg^?1) and theoretical ethanol yields, ETOHTL (L Mg^?1), based on cell wall and non-cell wall composition NIRS analyses. ETOHL means ranged from 98 to 115 L Mg^?1 while ETOHTL means ranged from 203 to 222 L Mg^?1. Because of differences in both biomass yields and conversion efficiency, there were significant differences among strains for both actual (2,534?C3,720 L ha^?1) and theoretical (4,878?C7,888 L ha^?1) ethanol production per hectare. It should be feasible to improve ethanol yields per hectare by improving both biomass yield and conversion efficiency by using NIRS analyses to quantify differences among cultivars and management practices.     

Mannitol production by heterofermentative <Emphasis Type="BoldItalic">Lactobacillus reuteri</Emphasis> CRL 1101 and <Emphasis Type="BoldItalic">Lactobacillus fermentum</Emphasis> CRL 573 in free and controlled pH batch fermentations

Certain lactic acid bacteria, especially heterofermentative strains, are capable to produce mannitol under adequate culture conditions. In this study, mannitol production by Lactobacillus reuteri CRL 1101 and Lactobacillus fermentum CRL 573 in modified MRS medium containing a mixture of fructose and glucose in a 6.5:1.0 ratio was investigated during batch fermentations with free pH and constant pH 6.0 and 5.0. Mannitol production and yields were higher under constant pH conditions compared with fermentations with free pH, the increase being more pronounced in the case of the L. fermentum strain. Maximum mannitol production and yields from fructose for L. reuteri CRL 1101 (122 mM and 75.7 mol%, respectively) and L. fermentum CRL 573 (312 mM and 93.5 mol%, respectively) were found at pH 5.0. Interestingly, depending on the pH conditions, fructose was used only as an alternative external electron acceptor or as both electron acceptor and energy source in the case of the L. reuteri strain. In contrast, L. fermentum CRL 573 used fructose both as electron acceptor and carbon source simultaneously, independently of the pH value, which strongly affected mannitol production by this strain. Studies on the metabolism of these relevant mannitol-producing lactobacilli provide important knowledge to either produce mannitol to be used as food additive or to produce it in situ during fermented food production.     

Adventitious Bud Formation from Bulb-scale Explants of Lilium speciosum Thunb. in vitro Effects of aminoethoxyvinyl-glycine, 1-aminocyclopropane-1-carboxylic acid,and ethylene

Several representatives of marine brown macroalgae (Phaeophyceae) including Fucus serratus L., Fucus spiralis L. and Fucus vesiculosus L. as well as Laminaria digitata (Huds.) Lamour., Laminaria hyperborea (Gunn.) Foslie and Laminaria saccharina (L.) Lamour. were investigated with particular regard to features of biosynthesis of the storage product mannitol. The respective catalytic system involved in the last step of mannitol formation, mannitol-1-phosphate dehydrogenase, appears to be a cytoplasmic enzyme as may be judged from the degree of correlation with the chloroplast key enzyme ribulose-1,5-bisphosphate carboxylase in different tissues of Laminaria digitata and Laminaria saccharina. Activity of mannitol-1-phosphate dehydrogenase in vitro is not affected by mannitol-l-phosphate or free mannitol, suggesting that mannitol biosynthesis in vivo) is mainly controlled by the environment and/or developmental stage. Certain inorganic ions such as NO₃^- (including K⁺) exert a strong influence on the activity of mannitol1-phosphate dehydrogenase thus suggesting that the intracellular pools of stored NO₃^- and mannitol are confined to spatially separated cellular compartments.     

Alginate degradation during anaerobic digestion of Laminaria hyperborea stipes

Polyphenols and divalent metal ions present in the tissue may seriously affect the degradation of alginate during anaerobic digestion of brown seaweeds. Laminaria hyperborea stipes, harvested at 59 °N off the Norwegian coast in the autumn, were degraded at different concentrations of polyphenols in anaerobic batch reactors at 35 °C and pH 7. This was done by removing or adding the mechanically peeled outer phenolic layer of the algae, and using methanogenic and alginate degrading inocula already adapted to L. hyperborea degradation. Initial alginate released from the algal particles was affected by NaOH titrations because the Ca/Na-ratio was reduced. After a rapid consumption of the mannitol, alginate lyases were induced, and guluronate lyases showed the highest extracellular activity. Then the microbes digested 0.12–0.23 g Na-alginate L⁻¹ h⁻¹. Later the degradation rate of alginates declined almost to zero, and 13–50% of the alginate remained insoluble. The total solubilisation of alginates was apparently limited by both Ca-crosslinked guluronate residues and complexation with compounds such as polyphenols. The methane production had a lag phase that increased at higher amounts of soluble polyphenols, and the total fermentation probably also became product inhibited if soluble compounds such as acetate, ethanol and butyrate were accumulated. This revised version was published online in September 2006 with corrections to the Cover Date.     

Kinetic modeling of Candida shehatae ATCC 22984 on xylose and glucose for ethanol production

Candida shehatae ATCC 22984, a xylose-fermenting yeast, showed an ability to produce ethanol in both glucose and xylose medium. Maximum ethanol produced by the yeast was 48.8?g/L in xylose and 52.6?g/L in glucose medium with ethanol yields that varied between 0.3 and 0.4?g/g depended on initial sugar concentrations. Xylitol was a coproduct of ethanol production using xylose as substrate, and glycerol was detected in both glucose and xylose media. Kinetic model equations indicated that growth, substrate consumption, and product formation of C. shehatae were governed by substrate limitation and inhibition by ethanol. The model suggested that cell growth was totally inhibited at 40?g/L of ethanol and ethanol production capacity of the yeast was 52?g/L, which were in good agreement with experimental results. The developed model could be used to explain C. shehatae fermentation in glucose and xylose media from 20 to 170?g/L sugar concentrations.     

Effect of carbon sources on the growth and ethanol production of native yeast Pichia kudriavzevii ITV-S42 isolated from sweet sorghum juice

The importance of non-Saccharomyces yeast species in fermentation processes is widely acknowledged. Within this group, Pichia kudriavzevii ITV-S42 yeast strain shows particularly desirable characteristics for ethanol production. Despite this fact, a thorough study of the metabolic and kinetic characteristics of this strain is currently unavailable. The aim of this work is to study the nutritional requirements of Pichia kudriavzevii ITV-S42 strain and the effect of different carbon sources on the growth and ethanol production. Results showed that glucose and fructose were both assimilated and fermented, achieving biomass and ethanol yields of 0.37 and 0.32 gg⁻¹, respectively. Glycerol was assimilated but not fermented; achieving a biomass yield of 0.88 gg⁻¹. Xylose and sucrose were not metabolized by the yeast strain. Finally, the use of a culture medium enriched with salts and yeast extract favored glucose consumption both for growth and ethanol production, improving ethanol tolerance reported for this genre (35 g L⁻¹) to 90 g L⁻¹ maximum ethanol concentration (over 100%). Furthermore Pichia kudriavzevii ITV-S42 maintained its fermentative capacity up to 200 g L⁻¹ initial glucose, demonstrating that this yeast is osmotolerant.

     

Enhancement of yeast ethanol tolerance by calcium and magnesium

Ethanol-induced changes of CO₂ production were compared in three strains ofSaccharomyces cerevisiœ. CaCl₂ and MgCl₂ exerted protective effects against the action of ethanol. Optimal concentrations ensuring maximum of CO₂ production at 10% (V/V) of ethanol under non-growing conditions were 3 mmol/L Ca²⁺ and 2 mmol/L Mg²⁺. Yeast growth with and without ethanol addition was stimulated by Mg²⁺ more than by Ca²⁺ during fermentation, whereas ethanol production was more efificient when both Ca²⁺ and Mg²⁺ were added.     

Optimization of fermentation parameters for production of ethanol from kinnow waste and banana peels by simultaneous saccharification and fermentation

A study was taken up to evaluate the role of some fermentation parameters like inoculum concentration, temperature, incubation period and agitation time on ethanol production from kinnow waste and banana peels by simultaneous saccharification and fermentation using cellulase and co-culture of Saccharomyces cerevisiae G and Pachysolen tannophilus MTCC 1077. Steam pretreated kinnow waste and banana peels were used as substrate for ethanol production in the ratio 4:6 (kinnow waste: banana peels). Temperature of 30°C, inoculum size of S. cerevisiae G 6% and (v/v) Pachysolen tannophilus MTCC 1077 4% (v/v), incubation period of 48 h and agitation for the first 24 h were found to be best for ethanol production using the combination of two wastes. The pretreated steam exploded biomass after enzymatic saccharification containing 63 gL⁻¹ reducing sugars was fermented with both hexose and pentose fermenting yeast strains under optimized conditions resulting in ethanol production, yield and fermentation efficiency of 26.84 gL⁻¹, 0.426 gg ⁻¹ and 83.52 % respectively. This study could establish the effective utilization of kinnow waste and banana peels for bioethanol production using optimized fermentation parameters.     

Consolidated bioprocessing strategy for ethanol production from Jerusalem artichoke tubers by Kluyveromyces marxianus under high gravity conditions

Aims: Developing an innovative process for ethanol fermentation from Jerusalem artichoke tubers under very high gravity (VHG) conditions. Methods and Results: A consolidated bioprocessing (CBP) strategy that integrated inulinase production, saccharification of inulin contained in Jerusalem artichoke tubers and ethanol production from sugars released from inulin by the enzyme was developed with the inulinase‐producing yeast Kluyveromyces marxianus Y179 and fed‐batch operation. The impact of inoculum age, aeration, the supplementation of pectinase and nutrients on the ethanol fermentation performance of the CBP system was studied. Although inulinase activities increased with the extension of the seed incubation time, its contribution to ethanol production was negligible because vigorously growing yeast cells harvested earlier carried out ethanol fermentation more efficiently. Thus, the overnight incubation that has been practised in ethanol production from starch‐based feedstocks is recommended. Aeration facilitated the fermentation process, but compromised ethanol yield because of the negative Crabtree effect of the species, and increases the risk of contamination under industrial conditions. Therefore, nonaeration conditions are preferred for the CBP system. Pectinase supplementation reduced viscosity of the fermentation broth and improved ethanol production performance, particularly under high gravity conditions, but the enzyme cost should be carefully balanced. Medium optimization was performed, and ethanol concentration as high as 94·2 g l^?1 was achieved when 0·15 g l^?1 K₂HPO₄ was supplemented, which presents a significant progress in ethanol production from Jerusalem artichoke tubers. Conclusions: A CBP system using K. marxianus is suitable for efficient ethanol production from Jerusalem artichoke tubers under VHG conditions. Significance and Impact of the Study: Jerusalem artichoke tubers are an alternative to grain‐based feedstocks for ethanol production. The high ethanol concentration achieved using K. marxianus with the CBP system not only saves energy consumption for ethanol distillation, but also significantly reduces the amount of waste distillage discharged from the distillation system.     

Performance comparison of ethanol and butanol production in a continuous and closed-circulating fermentation system with membrane bioreactor

Since both ethanol and butanol fermentations are urgently developed processes with the biofuel-demand increasing, performance comparison of aerobic ethanol fermentation and anerobic butanol fermentation in a continuous and closed-circulating fermentation (CCCF) system was necessary to achieve their fermentation characteristics and further optimize the fermentation process. Fermentation and pervaporation parameters including the average cell concentration, glucose consumption rate, cumulated production concentration, product flux, and separation factor of ethanol fermentation were 11.45?g/L, 3.70?g/L/h, 655.83?g/L, 378.5?g/m²/h, and 4.83, respectively, the corresponding parameters of butanol fermentation were 2.19?g/L, 0.61?g/L/h, 28.03?g/L, 58.56?g/m²/h, and 10.62, respectively. Profiles of fermentation and pervaporation parameters indicated that the intensity and efficiency of ethanol fermentation was higher than butanol fermentation, but the stability of butanol fermentation was superior to ethanol fermentation. Although the two fermentation processes had different features, the performance indicated the application prospect of both ethanol and butanol production by the CCCF system.     

Evaluation of the cellobiose-fermenting yeastBrettanomyces custersii in the simultaneous saccharification and fermentation of cellulose

Summary Brettanomyces custersii (CBS 5512) was identified as a promising glucose- and cellobiose-fermenting yeast for the simultaneous saccharification and fermentation (SSF) of cellulose for ethanol production. In SSF studies with 75 g/L of cellulose,B. custersii produced 32 g/L of ethanol in just 3 days (75% of theoretical yield). This yield represents an increase of more than 16% over the yields of other fementative yeasts and the time to achieve it is less than that with other organisms. In addition, the ethanol tolerance ofB. custersii seems to be greater than that of other cellobiose-fermenting yeasts considered to date. Overall, the combination of higher yields, rates, and ethanol concentrations obtained withB. custersii improves the economics of ethanol production.     

Regulation of nitric oxide production in snail (Lymnaea stagnalis) defence cells: a role for PKC and ERK signalling pathways

Background information. Nitric oxide (NO) is an important molecule in innate immune responses. In molluscs NO is produced by mobile defence cells called haemocytes; however, the molecular mechanisms that regulate NO production in these cells is poorly understood. The present study focused on the role of cell signalling pathways in NO production by primary haemocytes from the snail Lymnaea stagnalis. Results. When haemocytes were challenged with PMA (10 μM) or the β‐1,3‐glucan laminarin (10 mg/ml), an 8‐fold and 4‐fold increase in NO production were observed after 60 min respectively. Moreover, the NOS (NO synthase) inhibitors L‐NAME (N^G‐nitro‐L‐arginine methyl ester) and L‐NMMA (N^G‐monomethyl‐L‐arginine) were found to block laminarin‐ and PMA‐induced NO synthesis. Treatment of haemocytes with PMA or laminarin also increased the phosphorylation (activation) status of PKC (protein kinase C). When haemocytes were preincubated with PKC inhibitors (calphostin C or GF109203X) or inhibitors of the ERK (extracellular‐signal‐regulated kinase) pathway (PD98059 or U0126) prior to challenge, significant reductions in PKC and ERK phosphorylation and NO production were observed following exposure to laminarin or PMA. The greatest effect on NO production was seen with GF109203X and U0126, with PMA‐induced NO production inhibited by 94% and 87% and laminarin‐induced NO production by 50% and 91% respectively. Conclusions. These data suggest that ERK and PKC comprise part of the signalling machinery that regulates NOS activation and subsequent production of NO in molluscan haemocytes. To our knowledge, this is the first report that shows a role for these signalling proteins in the generation of NO in invertebrate defence cells.     

Contrasting genetic diversity patterns in two sister kelp species co‐distributed along the coast of Brittany,France

We investigated patterns of genetic structure in two sister kelp species to explore how distribution width along the shore, zonation, latitudinal distribution and historical factors contribute to contrasting patterns of genetic diversity. We implemented a hierarchical sampling scheme to compare patterns of genetic diversity and structure in these two kelp species co‐distributed along the coasts of Brittany (France) using a total of 12 microsatellites, nine for Laminaria hyperborea and 11 for Laminaria digitata, of which eight amplified in both species. The genetic diversity and connectivity of L. hyperborea populations were greater than those of L. digitata populations in accordance with the larger cross‐shore distribution width along the coast and the greater depth occupied by L. hyperborea populations in contrast to L. digitata populations. In addition, marginal populations showed reduced genetic diversity and connectivity, which erased isolation‐by‐distance patterns in both species. As L. digitata encounters its southern range limit in southern Brittany (SBr) while L. hyperborea extends down to mid‐Portugal, it was possible to distinguish the effect of habitat continuity from range edge effects. We found that L. digitata did not harbour high regional diversity at its southern edge, as expected in a typical rear edge, suggesting that refuges from the last glacial maximum for L. digitata were probably not located in SBr, but most likely further north. For both species, the highest levels of genetic diversity were found in the Iroise Sea and Morlaix Bay, the two regions in which they are being currently harvested. Preserving genetic diversity of these two foundation species in these areas should, thus, be a priority for the management of this resource in Brittany.