Progress towards a targeted biorefinery of Chromochloris zofingiensis: a review.

Biorefinery approaches offer the potential to improve the economics of the microalgae industry by producing multiple products from a single source of biomass. Chromochloris zofingiensis shows great promise for biorefinery due to high biomass productivity and a diverse range of products including secondary carotenoids, predominantly astaxanthin; lipids such as TAGs; carbohydrates including starch; and proteins and essential amino acids. Whilst this species has been demonstrated to accumulate multiple products, the development of an integrated downstream process to obtain these is lacking. The objective of this review paper is to assess the research that has taken place and to identify the steps that must be taken to establish a biorefinery approach for C. zofingiensis. In particular, the reasons why C. zofingiensis is a promising species to target for biorefinery are discussed in terms of cellular structure, potential products, and means to accumulate desirable components via the alteration of culture conditions. Future advances and the challenges that lie ahead for successful biorefinery of this species are also reviewed along with potential solutions to address them. Supplementary Information: The online version contains supplementary material available at 10.1007/s13399-022-02955-7.

A circular bioprocess application of algal-based substrate for Bacillus subtilis natto production of γ-PGA.

Poly-γ-glutamic acid (γ-PGA) is a bio-derived water-soluble, edible, hydrating, non-immunogenic polymer. Bacillus subtilis natto is a wild-type γ-PGA producer originally isolated from Japanese fermented natto beans whose activity has been shown to be enhanced through ion-specific activation of Extrachromosomal DNA maintenance mechanisms. Being a GRAS γ-PGA producer, this microorganism has attracted great interest in its use within an industrial context. Here we successfully synthesised amorphous, crystalline and semi-crystalline γ-PGA between 11-27 g/L. In line with circular economy principles, scalable macroalgal biomass has been evaluated as substrate for γ-PGA, displaying great potential in both yields and material composition. In this study whole cell, freeze dried seaweed -namely Laminaria digitata, Saccharina latissima and Alaria esculenta-were pre-treated by means of mechanical methods, sterilised and subsequently inoculated with B. subtilis natto. High shear mixing was found to be the most suitable pre-treatment technique. Supplemented L. digitata (9.1 g/L), S. latissima (10.2 g/L), A. esculenta (13 g/L) displayed γ-PGA yields comparable to those of standard GS media (14.4 g/L). Greatest yields of pure γ-PGA were obtained during the month of June for L. digitata (Avg. 4.76 g/L) comparable to those obtained with GS media (7.0 g/L). Further, pre-treated S. latissima and L. digitata complex media enabled for high molar mass (4,500 kDa) γ-PGA biosynthesis at 8.6 and 8.7 g/L respectively. Compared to standard GS media, algal derived γ-PGA displayed significantly higher molar masses. Further studies will be necessary to further evaluate the impact of varying ash contents upon the stereochemical properties and modify the properties of algal media based γ-PGA with the aid of key nutrients; however, the material synthesised to date can directly displace a number of fossil fuel derived chemicals in drug delivery applications, cosmetics, bioremediation, wastewater treatment, flocculation and as cryoprotectants.

Microbial Poly-γ-Glutamic Acid (γ-PGA) as an Effective Tooth Enamel Protectant.

Poly-γ-glutamic acid (γ-PGA) is a bio-derived water-soluble, edible, non-immunogenic nylon-like polymer with the biochemical characteristics of a polypeptide. This Bacillus-derived material has great potential for a wide range of applications, from bioremediation to tunable drug delivery systems. In the context of oral care, γ-PGA holds great promise in enamel demineralisation prevention. The salivary protein statherin has previously been shown to protect tooth enamel from acid dissolution and act as a reservoir for free calcium ions within oral cavities. Its superb enamel-binding capacity is attributed to the L-glutamic acid residues of this 5380 Da protein. In this study, γ-PGA was successfully synthesised from Bacillus subtilis natto cultivated on supplemented algae media and standard commercial media. The polymers obtained were tested for their potential to inhibit demineralisation of hydroxyapatite (HAp) when exposed to caries simulating acidic conditions. Formulations presenting 0.1, 0.25, 0.5, 0.75, 1, 2, 3 and 4% (w/v) γ-PGA concentration were assessed to determine the optimal conditions. Our data suggests that both the concentration and the molar mass of the γ-PGA were significant in enamel protection (p = 0.028 and p < 0.01 respectively). Ion Selective Electrode, combined with Fourier Transform Infra-Red studies, were employed to quantify enamel protection capacity of γ-PGA. All concentrations tested showed an inhibitory effect on the dissolution rate of calcium ions from hydroxyapatite, with 1% (wt) and 2% (wt) concentrations being the most effective. The impact of the average molar mass (M) on enamel dissolution was also investigated by employing commercial 66 kDa, 166 kDa, 440 kDa and 520 kDa γ-PGA fractions. All γ-PGA solutions adhered to the surface of HAp with evidence that this remained after 60 min of continuous acidic challenge. Inductively Coupled Plasma analysis showed a significant abundance of calcium ions associated with γ-PGA, which suggests that this material could also act as a responsive calcium delivery system. We have concluded that all γ-PGA samples tested (commercial and algae derived) display enamel protection capacity regardless of their concentration or average molar mass. However, we believe that γ-PGA D/L ratios might affect the binding more than its molar mass.

Quality and Safety Assessment of Edible Seaweeds Alaria esculenta and Saccharina latissima Cultivated in Scotland.

Within Europe over the last 10 years, there has been an increase in seaweeds cultivated for human consumption. For food safety reasons, it is important to assess the microbiological and nutritional quality of the biomass. The fresh and dried edible seaweeds Alaria esculenta and Saccharina latissima were assessed over two consecutive years for the presence of microorganisms. Seaweed samples supplied from Scotland were stored under isothermal conditions for specific time intervals depending on the sample's condition (fresh, dried or rehydrated). During storage, microbiological analyses were performed for the enumeration of Total Viable Counts (TVC), Pseudomonas spp., Enterobacteriaceae and Bacillus spp., as well as yeasts and molds. Additionally, bacterial colonies from the Marine Agar growth medium were isolated and subjected to PCR-RAPD analysis for characterization of the bacterial diversity of seaweeds. Bacterial isolates with different fingerprint patterns were further subjected to sequencing (16S rDNA, V1-V4 region). The presence of human pathogenic bacteria was also investigated. Results showed that the initial population of TVC was differentiated depending on the year of seaweed harvest, being closer to the enumeration limit (1.0 log CFU/g) in fresh samples from 2020 and higher in samples from 2019 (6.7 and 3.9 log CFU/g in A. esculenta and S. latissima, respectively). DNA-based analysis revealed the presence of Psychrobacter, Cobetia and Pseudomonas species in A. esculenta, while Psychrobacter and Micrococcus species were present in S. latissima.

Enabling large-scale production of algal oil in continuous output mode.

Large-scale algal oil production requires continuous outputs and a trade-off between growth and oil content. Two unrelated marine algae (Nannochloropsis oceanica [CCAP 849/10] and Chlorella vulgaris [CCAP 211/21A]) that showed high oil production under batch culture were studied under controlled semicontinuous cultivation conditions. Three essential attributes maximized oil productivity: (i) downregulation of cell size to maximize light absorption under N limitation; (ii) low nutrient-depletion thresholds to trigger oil induction; (iii) a means of carbohydrate suppression in favor of oil. N. oceanica responded better to input N/P variations and is more suited to continuous oil production. A low N/P ratio was effective in both suppressing carbohydrate and reducing cell size concomitant with oil production. In C. vulgaris, nutrient starvation thresholds for oil were higher and carbohydrate was preferentially induced, which impeded stress-level optimization for oil. These differences, which impact continuous oil production at scale, are driven by species adaptation to specific marine habitats.

Charting a course for genetic diversity in the UN Decade of Ocean Science.

The health of the world's oceans is intrinsically linked to the biodiversity of the ecosystems they sustain. The importance of protecting and maintaining ocean biodiversity has been affirmed through the setting of the UN Sustainable Development Goal 14 to conserve and sustainably use the ocean for society's continuing needs. The decade beginning 2021-2030 has additionally been declared as the UN Decade of Ocean Science for Sustainable Development. This program aims to maximize the benefits of ocean science to the management, conservation, and sustainable development of the marine environment by facilitating communication and cooperation at the science-policy interface. A central principle of the program is the conservation of species and ecosystem components of biodiversity. However, a significant omission from the draft version of the Decade of Ocean Science Implementation Plan is the acknowledgment of the importance of monitoring and maintaining genetic biodiversity within species. In this paper, we emphasize the importance of genetic diversity to adaptive capacity, evolutionary potential, community function, and resilience within populations, as well as highlighting some of the major threats to genetic diversity in the marine environment from direct human impacts and the effects of global climate change. We then highlight the significance of ocean genetic diversity to a diverse range of socioeconomic factors in the marine environment, including marine industries, welfare and leisure pursuits, coastal communities, and wider society. Genetic biodiversity in the ocean, and its monitoring and maintenance, is then discussed with respect to its integral role in the successful realization of the 2030 vision for the Decade of Ocean Science. Finally, we suggest how ocean genetic diversity might be better integrated into biodiversity management practices through the continued interaction between environmental managers and scientists, as well as through key leverage points in industry requirements for Blue Capital financing and social responsibility.

Removal of metals from aqueous solutions using dried Cladophora parriaudii of varying biochemical composition.

Macroalgal biosorption has shown promise for the removal of metal ions from wastewaters, whose presence can pose a threat to the aquatic environment. There is a wealth of literature published on macroalgal biosorption, the common thread being that the biosorbent material was collected from the field, under undefined conditions. These studies offer little insight into the impact of prior cultivation or biomass production practices upon the biosorbent material, its adsorptive physico-chemical properties and its subsequent capacity for metal removal. The present study sought to investigate the influence of changes in macroalgal cultivation, specifically nutrient regime, upon biomass properties and the resultant adsorption performance. The macroalga Cladophora parriaudii was cultivated under six different nutrient regimes; 2:1 and 12:1 N:P molar ratios, with nitrogen supplied either as ammonium (NH4+), nitrate (NO3-), or urea (CO(NH2)2). These nutrient regimes were designed to produce biomass of varying biochemical and cell surface profiles. After cultivation, the biomass was rinsed, dried, biochemically analysed and then used for the removal of four individual metals from solution. Metal removal varied considerably between treatments and across initial metal concentrations, with removal values of 46-85%, 9-80%, 8-71%, and 49-94% achieved for Al, Cu, Mn, and Pb, respectively, with initial metal concentrations varying between 0 and 150 mg L-1. The observed variation in metal removal can only be attributed to differences in biochemistry and cell surface properties of the biosorbent induced by nutrient regime, as all other variables were constant. This study demonstrates that prior cultivation conditions influence the biochemistry of a biosorbent material, namely macroalgae Cladophora parriaudii, which has an impact upon metal removal. This aspect should be given due consideration for future biosorption research and when reviewing already published literature.

Exploring the Chemical Space of Macro- and Micro-Algae Using Comparative Metabolomics.

With more than 156,000 described species, eukaryotic algae (both macro- and micro-algae) are a rich source of biological diversity, however their chemical diversity remains largely unexplored. Specialised metabolites with promising biological activities have been widely reported for seaweeds, and more recently extracts from microalgae have exhibited activity in anticancer, antimicrobial, and antioxidant screens. However, we are still missing critical information on the distinction of chemical profiles between macro- and microalgae, as well as the chemical space these metabolites cover. This study has used an untargeted comparative metabolomics approach to explore the chemical diversity of seven seaweeds and 36 microalgal strains. A total of 1390 liquid chromatography-mass spectrometry (LC-MS) features were detected, representing small organic algal metabolites, with no overlap between the seaweeds and microalgae. An in-depth analysis of four Dunaliella tertiolecta strains shows that environmental factors may play a larger role than phylogeny when classifying their metabolomic profiles.

Comparing the effectiveness of twine- and binder-seeding in the Laminariales species Alaria esculenta and Saccharina latissima.

The continuing expansion of seaweed cultivation could assist in ensuring future global food security. The Laminariales species Alaria esculenta and Saccharina latissima are each cultivated for food across their European ranges. The predominant method for cultivating European kelps involves growing juveniles on twine within a hatchery which is then deployed at a farm site. The associated hatchery and deployment cost of this approach are relatively high. A new and innovative methodology-called binder-seeding-can reduce these costs, but, has yet to be validated. We compare the biomass yield and morphology of A. esculenta and S. latissima cultured using either the traditional twine-longline method or binder-seeding onto AlgaeRope and AlgaeRibbon, specially designed textiles. In a controlled growth experiment, A. esculenta had a similar biomass yield on all materials, but fronds were shorter (23 ± 7%) and thinner on the AlgaeRibbon (42 ± 4%) due to a 3-4-fold higher density of developing sporophytes compared to the twine-longline. In contrast, S. latissima gave a 4-fold higher biomass yield on the AlgaeRibbon in June (4.0 kg m-1), but frond morphology was not different between materials, despite a 4-fold higher sporophyte density on the AlgaeRibbon. The stipe length of both species also increased at the higher sporophyte density on the AlgaeRibbon. The AlgaeRope gave an intermediate response or was similar to the twine-longline. These results show that binder-seeding onto the AlgaeRibbon significantly increases the achieved biomass yield in S. latissima. These results can assist cultivators to select the most appropriate method of kelp cultivation depending on morphological/yield requirements of the end use. Further study is needed on the optimisation of the binder-seeding density and its impact on thallus morphology.

Lipidomics analysis of juveniles' blue mussels (Mytilus edulis L. 1758), a key economic and ecological species.

Blue mussels (Mytilus edulis L. 1758) are important components of coastal ecosystems and in the economy of rural and coastal areas. The understanding of their physiological processes at key life stages is important both within food production systems and in the management of wild populations. Lipids are crucial molecules for bivalve growth, but their diversity and roles have not been fully characterised. In this study, traditional lipid profiling techniques, such as fatty acid (FA) and lipid class analysis, are combined to untargeted lipidomics to elucidate the lipid metabolism in newly settled spat fed on a range of diets. The evaluated diets included single strains treatments (Cylindrotheca fusiformis CCAP 1017/2 -CYL, Isochrysis galbana CCAP 927/1- ISO, Monodopsis subterranean CCAP 848/1 -MONO, Nannochloropsis oceanica CCAP 849/10- NANNO) and a commercial algae paste (SP). Spat growth was influenced by the diets, which, according to their efficacy were ranked as follows: ISO>NANNO/CYL>SP>MONO. A higher triacylglycerols (TG) content, ranging from 4.23±0.82 µg mgashfree Dry weight (DW)-1 at the beginning of the trial (T0) to 51±15.3 µg mgashfreeDW-1 in ISO, characterised significant growth in the spat, whereas, a reduction of TG (0.3±0.08 µg mgashfreeDW-1 in MONO), mono unsaturated FA-MUFA (from 8.52±1.02 µg mgFAashfreeDW-1 at T0 to 2.81±1.02 µg mgFAashfreeDW-1 in MONO) and polyunsaturated FA-PUFA (from 17.57±2.24 µg mgFAashfreeDW-1 at T0 to 6.19±2.49 µg mgFAashfreeDW-1 in MONO) content characterised poor performing groups. Untargeted lipidomics evidenced how the availability of dietary essential PUFA did not influence only neutral lipids but also the membrane lipids, with changes in lipid molecular species in relation to the essential PUFA provided via the diet. Such changes have the potential to affect spat production cycle and their ability to respond to the surrounding environment. This study evidenced the advantages of coupling different lipid analysis techniques, as each technique disclosed relevant information on nutritional requirements of M. edulis juveniles, expanding the existing knowledge on the physiology of this important species.

Microalgal Carotenoids: A Review of Production, Current Markets, Regulations, and Future Direction.

Microalgae produce a variety of compounds that are beneficial to human and animal health. Among these compounds are carotenoids, which are microalgal pigments with unique antioxidant and coloring properties. The objective of this review is to evaluate the potential of using microalgae as a commercial feedstock for carotenoid production. While microalgae can produce some of the highest concentrations of carotenoids (especially astaxanthin) in living organisms, there are challenges associated with the mass production of microalgae and downstream processing of carotenoids. This review discusses the synthesis of carotenoids within microalgae, their physiological role, large-scale cultivation of microalgae, up- and down-stream processing, commercial applications, natural versus synthetic carotenoids, and opportunities and challenges facing the carotenoid markets. We emphasize legal aspects and regulatory challenges associated with the commercial production of microalgae-based carotenoids for food/feed, nutraceutical and cosmetic industry in Europe, the USA, the People's Republic of China, and Japan. This review provides tools and a broad overview of the regulatory processes of carotenoid production from microalgae and other novel feedstocks.

Effects of cryopreservation on viability and functional stability of an industrially relevant alga.

As algal biotechnology develops, there is an increasing requirement to conserve cultures without the cost, time and genetic stability implications of conventional serial transfers, including issues regarding potential loss by failure to regrow, contamination on transfer, mix up and/or errors in the documentation on transfer. Furthermore, it is crucial to ensure both viability and functionality are retained by stored stock-cultures. Low temperature storage, ranging from the use of domestic freezers to storage under liquid nitrogen, is widely being used, but the implication to stability and function rarely investigated. We report for the first time, retention of functionality in the maintenance of master stock-cultures of an industrially relevant, lipid-producing alga, under a variety of cryopreservation regimes. Storage in domestic (-15 °C), or conventional -80 °C freezers was suboptimal, with a rapid reduction in viability observed for samples at -15 °C and a >50% loss of viability, within one month, for samples stored at -80 °C. No reduction in viability occurred at -196 °C. Post-thaw culture functional performance was also influenced by the cryopreservation approach employed. Only samples held at -196 °C responded to nitrogen limitation in terms of growth characteristics and biochemical profiles (lipid production and chlorophyll a) comparable to the untreated control, cultured prior to cryopreservation. These results have important implications in microbial biotechnology, especially for those responsible for the conservation of genetic resources.

A comparison of methods for the non-destructive fresh weight determination of filamentous algae for growth rate analysis and dry weight estimation.

The determination of rates of macroalgal growth and productivity via temporal fresh weight (FW) measurements is attractive, as it does not necessitate the sacrifice of biomass. However, there is no standardised method for FW analysis; this may lead to potential discrepancies when determining growth rates or productivity and make literature comparison problematic. This study systematically assessed a variety of lab-scale methods for macroalgal FW measurement for growth rate determination. Method efficacy was assessed over a 14-day period as impact upon algal physiology, growth rate on basis of FW and dry weight (DW), nitrate removal, and maintenance of structural integrity. The choice of method is critical to both accuracy and inter-study comparability of the data generated. In this study, it was observed that the choice of protocol had an impact upon the DW yield (P values = 0.036-0.51). For instance, those involving regular mechanical pressing resulted in a >25% reduction in the final DW in two of the three species studied when compared to algae not subjected to any treatment. This study proposes a standardised FW determination method employing a reticulated spinner that is rapid, reliable, and non-destructive and provides an accurate growth estimation.

Media Screening for Obtaining Haematococcus pluvialis Red Motile Macrozooids Rich in Astaxanthin and Fatty Acids.

Astaxanthin from Haematococcus pluvialis is commercially produced in a two-stage process, involving green vegetative (macrozooid) and red aplanospore stages. This approach has been scaled up to an industrial process but constraints limit its commercial success and profitability, including: contamination issues, high pigment extraction costs, requirements for high light levels and photo-bleaching in the red stage. However, in addition to the aplanospore stage, this alga can produce astaxanthin in vegetative palmelloid and motile macrozooid cells. In this study, a two-stage process utilising different media in the green stage, with subsequent re-suspension in medium without nitrate was employed to optimise the formation of red motile macrozooids. Optimal growth in the green phase was obtained on cultivation under mixotrophic conditions in EG:JM media followed by re-suspension in medium without nitrate resulting in red motile macrozooids with an astaxanthin content of 2.74% (78.4% of total carotenoids) and a lipid content of 35.3% (rich in unsaturated fatty acids. It is envisaged that the red motile macrozooids could be harvested and fed as a whole-cell product directly in the animal feed and aquaculture sectors, or used as a blend of carotenoids and polyunsaturated fatty acids (PUFAs) in nutraceutical products.

Biofouling community composition across a range of environmental conditions and geographical locations suitable for floating marine renewable energy generation.

Knowledge of biofouling typical of marine structures is essential for engineers to define appropriate loading criteria in addition to informing other stakeholders about the ecological implications of creating novel artificial environments. There is a lack of information regarding biofouling community composition (including weight and density characteristics) on floating structures associated with future marine renewable energy generation technologies. A network of navigation buoys were identified across a range of geographical areas, environmental conditions (tidal flow speed, temperature and salinity), and deployment durations suitable for future developments. Despite the perceived importance of environmental and temporal factors, geographical location explained the greatest proportion of the observed variation in community composition, emphasising the importance of considering geography when assessing the impact of biofouling on device functioning and associated ecology. The principal taxa associated with variation in biofouling community composition were mussels (Mytilus edulis), which were also important when determining loading criteria.

Unlocking nature's treasure-chest: screening for oleaginous algae.

Micro-algae synthesize high levels of lipids, carbohydrates and proteins photoautotrophically, thus attracting considerable interest for the biotechnological production of fuels, environmental remediation, functional foods and nutraceuticals. Currently, only a few micro-algae species are grown commercially at large-scale, primarily for "health-foods" and pigments. For a range of potential products (fuel to pharma), high lipid productivity strains are required to mitigate the economic costs of mass culture. Here we present a screen concentrating on marine micro-algal strains, which if suitable for scale-up would minimise competition with agriculture for water. Mass-Spectrophotometric analysis (MS) of nitrogen (N) and carbon (C) was subsequently validated by measurement of total fatty acids (TFA) by Gas-Chromatography (GC). This identified a rapid and accurate screening strategy based on elemental analysis. The screen identified Nannochloropsis oceanica CCAP 849/10 and a marine isolate of Chlorella vulgaris CCAP 211/21A as the best lipid producers. Analysis of C, N, protein, carbohydrate and Fatty Acid (FA) composition identified a suite of strains for further biotechnological applications e.g. Dunaliella polymorpha CCAP 19/14, significantly the most productive for carbohydrates, and Cyclotella cryptica CCAP 1070/2, with utility for EPA production and N-assimilation.

Comparison of screening methods for high-throughput determination of oil yields in micro-algal biofuel strains.

The phenotypic and phylogenetic diversity of micro-algae capable of accumulating triacylglycerols provides a challenge for the accurate determination of biotechnological potential. High-yielding strains are needed to improve economic viability and their compositional information is required for optimizing biodiesel properties. To facilitate a high-throughput screening programme, a very rapid direct-derivatization procedure capable of extracting lyophilized material for GC analysis was compared with a scaled-down Folch-based method. This was carried out on ten micro-algal strains from 6 phyla where the more rapid direct-derivatization approach was found to provide a more reliable measure of yield. The modified Folch-based procedure was found to substantially underestimate oil yield in one Chlorella species (P < 0.01). In terms of fatty acid composition however, the Folch procedure proved to be slightly better in recovering polyunsaturated fatty acids, in six out of the ten strains. Therefore, direct-derivatization is recommended for rapid determination of yields in screening approaches but can provide slightly less compositional accuracy than solvent-based extraction methods.

Effects of temperature and nutrient regimes on biomass and lipid production by six oleaginous microalgae in batch culture employing a two-phase cultivation strategy.

Commercial success of algal-based biofuels depends on growth characteristics and lipid metabolism of the production species. The oleaginous microalgae, Thalassiosira pseudonana, Odontella aurita, Nannochloropsis oculata, Isochrysis galbana, Chromulina ochromonoides, and Dunaliella tertiolecta, were cultivated under a matrix of two temperatures (10 and 20 °C) and two nutrient regimes (deplete and replete). For all species, a strong negative correlation between growth rate and lipid content was observed. Multiple stressors have no additive effect on lipid accumulation. Total oil content (fatty acid methyl esters, FAMEs, pg cell(-1)) was increased more by nutrient limitation than by temperature stress. In response to nutrient stress, N. oculata emerged as the most robust species with an increase in lipid accumulation of up to three to four-fold compared to the accumulation under nutrient sufficient conditions. Although stress conditions led to reduced fatty acid unsaturation in most taxa due to increased triacylglycerol (TAG) production, a high proportion of eicosapentaenoic acid (EPA) was maintained in O. aurita.

A rapid and general method for measurement of protein in micro-algal biomass.

A convenient small-scale extraction method for lyophilized micro-algae is described that dispenses with labor-intensive homogenization and is widely applicable to algae from different phyla. The procedure employs an optimized sequential extraction in trichloroacetic acid (TCA) and NaOH to achieve chemical lysis. Conditions were tested using several micro-algal strains to develop a method that was generally applicable. Incubation of lyophilized material in 24% (w/v) TCA at 95 °C followed by a hot alkaline treatment was found to be effective for strains that are resistant to conventional extraction approaches, such as the Chlorella and the Eustigmatophycean species. The single-tube extraction procedure can be complete in 4h and is conveniently followed by the Lowry assay, requiring a further 30 min. Overall, this method proved to be generally applicable and ideal either for single samples or for high-throughput screening of multiple algal strains for protein content.

Biogas from Macroalgae: is it time to revisit the idea?

The economic and environmental viability of dedicated terrestrial energy crops is in doubt. The production of large scale biomass (macroalgae) for biofuels in the marine environment was first tested in the late 1960's. The culture attempts failed due to the engineering challenges of farming offshore. However the energy conversion via anaerobic digestion was successful as the biochemical composition of macroalgae makes it an ideal feedstock. The technology for the mass production of macroalgae has developed principally in China and Asia over the last 50 years to such a degree that it is now the single largest product of aquaculture. There has also been significant technology transfer and macroalgal cultivation is now well tried and tested in Europe and America. The inherent advantage of production of biofuel feedstock in the marine environment is that it does not compete with food production for land or fresh water. Here we revisit the idea of the large scale cultivation of macroalgae at sea for subsequent anaerobic digestion to produce biogas as a source of renewable energy, using a European case study as an example.