Bio-processing of macroalgae Palmaria palmata: metabolite fractionation from pressed fresh material and ensiling considerations for long-term storage.

Red algae, belonging to the phylum Rhodophyta, contain an abundance of useful chemicals including bioactive molecules and present opportunities for the production of different products through biorefinery cascades. The rhodophyte Palmaria palmata, commonly termed dulse or dillisk, grows predominantly on the northern coasts of the Atlantic and Pacific Oceans and is a well-known snack food. Due to its abundance, availability and cultivation capacity, P. palmata was selected for study as a potential candidate for a biorefinery process. In addition to studying juice and solid fractions of freshly harvested P. palmata, we have investigated the novel possibility of preserving algal biomass by ensilaging protocols similar to those employed for terrestrial forage crops. In the metabolite partitioning within the solid and liquid fractions following screw-pressing, the majority of the metabolites screened for-water soluble carbohydrates, proteins and amino acids, lipids, pigments, phenolics and antioxidant activity-remained in the solid fraction, though at differing proportions depending on the metabolite, from 70.8% soluble amino acids to 98.2% chlorophyll a and 98.1% total carotenoids. For the ensiling study, screw-pressed P. palmata, with comparative wilted and chopped, and chopped only samples, were ensiled at scale with and without Safesil silage additive. All samples were successfully ensiled after 90 days, with screw-pressing giving lower or equal pH before and after ensiling compared with the other preparations. Of particular note was the effluent volumes generated during ensiling: 26-49% of the fresh weight, containing 16-34% of the silage dry matter. This may be of advantage depending on the final use of the biomass.

What cell wall components are the best indicators for Miscanthus digestibility and conversion to ethanol following variable pretreatments?

BACKGROUND: Energy crops including Miscanthus provide a storable, portable energy source which can be used to complement a wide range of products and energy generation systems. Miscanthus is predominantly used in Europe as a combustion material for electricity generation but also has the potential for biochemical conversion due to its high yield and low-nutrient requirements. The ratio of holocellulose (hemicellulose and cellulose combined) to acid detergent lignin (H:L) within the senesced material has previously been shown to indicate the relative suitability of Miscanthus accessions for thermochemical conversion. In this study, the ratio was assessed to examine its use as a selection aid for biochemical conversion. 20 highly-characterised Miscanthus accessions were saccharified using an enzyme mix to determine optimum sugar release. Nine of these accessions spanning high, medium and low H:L ratios were then autoclaved with dilute acid, alkali or water, and enzymically hydrolysed and fermented to produce ethanol. Samples taken throughout the process allowed assessments of released sugars. RESULTS: Enzymic degradation of the biomass showed a relationship between H:L ratio and glucose release, with high glucose release for high H:L ratio accessions and vice versa. Xylose release showed no such relationship. This relationship was maintained following pretreatments and enzyme saccharification, where compound analysis showed that following all pretreatments, accessions with high H:L ratios repeatedly had the highest releases of glucose, xylose and arabinose, and produced more ethanol. Release of all measured compounds increased with the pretreatment severity and ethanol yields from each pretreatment correlated with the respective glucose yield, providing assurance that any inhibitory compounds generated were tolerated by the fermentation yeast. Strong correlations were also seen between glucose release, ethanol and cell wall components, with cellulose showing the highest correlations with ethanol yields for some treatments and H:L ratio with others. CONCLUSIONS: The H:L ratio is a good predictor of ethanol yields and sugar release from Miscanthus in this study but individual components lignin and cellulose also correlate well, especially for hot water and mild acid pretreatments. In conclusion, use of the H:L ratio does not provide any advantages over the concentration of individual cell wall components for predicting sugar release and ethanol yields.

Novel rapid method for the characterisation of polymeric sugars from macroalgae.

Laminarins are storage polysaccharides found only in brown seaweeds, specifically Laminarialaes and Fucales. Laminarin has been shown to have anti-apoptotic and anti-tumoural activities and is considered as a nutraceutical component that can positively influence human health. The structure is species dependent, generally composed of linear ß(1-3) glucans with intrachain ß(1-6) branching and varies according to harvest season and environmental factors. Current methods for analysis of molar mass and DP length are technically demanding and are not widely available. Here, we present a simple inexpensive method which enables rapid analysis of laminarins from macroalgal biomass using high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) without the need for hydrolysis or further processing. This is based on the linear relationship observed between log10 DP and retention time following separation of laminarins on a CarboPac PA-100 column (Dionex) using standard 1,3-ß-d-gluco-oligosaccharides ranging in DP from 2 to 8. This method was applied to analyse laminarin oligomers in extracts from different species harvested from within the intertidal zone on Welsh rocky shores containing laminarin polymers with different ranges of DP. The degree of polymerisation and extrapolated molar mass agreed well with values estimated by LC-ESI/MS n analysis and those reported in the literature.

Seasonal variation in Laminaria digitata and its impact on biochemical conversion routes to biofuels.

Laminaria digitata is a highly prevalent kelp growing off the coast of the UK but has rarely been considered as a source of biomass to date. This study shows it can be used as a feedstock in both ethanol fermentation and anaerobic digestion for methane production. The study optimised several parameters in the fermentation of L. digitata and investigated the suitability of the macroalgae through the year using samples harvested every month. For both methane and ethanol production, minimum yields were seen in material harvested in March when the carbohydrates laminarin and mannitol were lowest. July material contained the highest combined laminarin and mannitol content and maximum yields of 167 mL ethanol and 0.219 m(3) kg(-1)L. digitata.

Seasonal variation in the chemical composition of the bioenergy feedstock Laminaria digitata for thermochemical conversion.

To avoid negative impacts on food production, novel non-food biofuel feedstocks need to be identified and utilised. One option is to utilise marine biomass, notably fast-growing, large marine 'plants' such as the macroalgal kelps. This paper reports on the changing composition of Laminaria digitata throughout it growth cycle as determined by new technologies. The potential of Laminaria sp. as a feedstock for biofuel production and future biorefining possibilities was assessed through proximate and ultimate analysis, initial pyrolysis rates using thermo-gravimetric analysis (TGA), metals content and pyrolysis gas chromatography-mass spectrometry. Samples harvested in March contained the lowest proportion of carbohydrate and the highest ash and alkali metal content, whereas samples harvested in July contained the highest proportions of carbohydrate, lowest alkali metals and ash content. July was therefore considered the most suitable month for harvesting kelp biomass for thermochemical conversion to biofuels.