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As an eco-friendly energy source, photobioreactors are drawing high interest. In these reactors, phototrophic microorganisms such as microalgae harness solar energy to produce biomass, making the amount of solar energy the most critical factor for the reactors' productivity. In this paper, we present a method to determine the optimal arrangement of photobioreactor panels in an array to maximize the accumulation of solar energy over a specified period. The panels are flat, rectangular, and vertically arrayed with uniform spacing. This arrangement results in the shading of direct solar beams between adjacent panels and illumination from diffuse radiation. We calculate the shaded area, which varies with time, and estimate the illumination effect between panels by determining the solid angle of diffused light sources. We begin by predicting the solar position and local frame vectors in the ecliptic frame using our numerical model and then using the solar radiation model to calculate the accumulated solar energy on the reactor panels. Our optimization goal is efficiency, which balances productivity and installation costs. Installation cost includes the panel price and land value and affects the reactor efficiency, which depends on panel spacing and land area. We use dimensionless efficiency, where all lengths are expressed as ratios to the panel height. Finally, we find the optimal dimensionless spacing and panel tilt angle to provide maximum productivity over the cost at selected locations specified by latitude and longitude.

During Drosophila embryogenesis, the essential pioneer factor Zelda defines hundreds of cis-regulatory regions and in doing so reprograms the zygotic transcriptome. While Zelda is essential later in development, it is unclear how the ability of Zelda to define cis-regulatory regions is shaped by cell-type-specific chromatin architecture. Asymmetric division of neural stem cells (neuroblasts) in the fly brain provide an excellent paradigm for investigating the cell-type-specific functions of this pioneer factor. We show that Zelda synergistically functions with Notch to maintain neuroblasts in an undifferentiated state. Zelda misexpression reprograms progenitor cells to neuroblasts, but this capacity is limited by transcriptional repressors critical for progenitor commitment. Zelda genomic occupancy in neuroblasts is reorganized as compared to the embryo, and this reorganization is correlated with differences in chromatin accessibility and cofactor availability. We propose that Zelda regulates essential transitions in the neuroblasts and embryo through a shared gene-regulatory network driven by cell-type-specific enhancers., The pioneer transcription factor Zelda is essential for the reprogramming of germ cells to totipotent cells of the early Drosophila embryo. Here the authors examine the function of Zelda later in development in the larval brain to show that Zelda promotes the undifferentiated stem-cell fate. They further identify factors that limit the reprogramming capacity of this pioneer factor.

Synechocystis sp. PCC 7338 (hereafter referred to as Synechocystis 7338) is a marine cyanobacterium that has the potential to produce photosynthetic pigments. In this study, we investigated the effects of various NaCl concentrations (0, 0.4, 0.8, and 1.2 M) on cell growth, photosynthetic pigments, and metabolites and intact lipid species profiles in Synechocystis 7338. The overall growth pattern of Synechocystis 7338 was similar under 0, 0.4, and 0.8 M NaCl conditions. Cell growth was retarded after reaching the exponential phase under 1.2 M NaCl; however, a similar growth pattern was observed after the exponential phase under 0.4 M NaCl (control group). The highest production of chlorophyll a (4.18 mg L−1), allophycocyanin (4.08 mg L−1), and phycoerythrin (1.70 mg L−1) were achieved under 1.2 M NaCl conditions. Altered metabolic and lipidomic profiles were observed at different NaCl conditions; significantly increased relative yields of glucosylglycerol, one diacylglyceryltrimethylhomoserine, one monogalactosyldiacylglycerol, and four phosphatidylglycerol species were observed under 1.2 M NaCl conditions using gas chromatography–mass spectrometry and direct infusion–mass spectrometry analyses. In addition, it was revealed that the photosynthetic activity recovered under 1.2 M NaCl conditions in long-term culture. Hydrogen peroxide content significantly increased under 1.2 M NaCl conditions. It is believed that glutathione content also significantly increased under high salinity conditions to retain the normal functioning of Synechocystis 7338. These results indicate that high salinity conditions for Synechocystis 7338 culture could be used for the large-scale production of chlorophyll a, allophycocyanin, phycoerythrin, and other bioactive metabolites.

This paper presents a new surface modification strategy to develop a poly(ethylene terephthalate) (PET)-based membrane having a hydrophilic surface, high nutrient ion permeability, sufficient mechanical strength, and organic fouling resistance, using an anthracene (ANT)-attached polyethylene glycol (PEG) surface modification agent (SMA) synthesized in this work. During the modification process, the ANT parts of the SMAs poke through and anchor to the surface of a commercial PET woven fabric via physical interactions and mechanical locking. The PEG chain parts coat the surface in the brush and arch forms, which generates a hydration layer on the fabric surface. The consequently obtained surface property and unique structure of the modified PET-based membrane result in higher nitrate ion permeability, organic fouling resistance, and microalgae production compared to those of the unmodified one. These are also affected by the molecular weight of the PEG and the number density of the anchored SMAs. The study demonstrates that this new surface modification method has the potential to allow the development of a desirable PET-based membrane for the efficient massive production of marine microalgae.

Synechocystis strains are considered as valuable resources for production of useful metabolites. Comparative primary metabolic and lipidomic profiling of freshwater and marine Synechocystis strains is needed to understand the physiological characteristics of each strain at molecular level. In this study, we performed comparative primary metabolic and lipidomic profiling of freshwater Synechocystis sp. PCC 6803 (hereafter Synechocystis 6803) and marine Synechocystis sp. PCC 7338 (hereafter Synechocystis 7338) by using gas chromatography-mass spectrometry and nano electrospray ionization-mass spectrometry. Neophytadiene, phytol, monogalactosyldiacylglycerol (MGDG), and digalactosyldiacylglycerol (DGDG) were significantly higher in Synechocystis 6803. Levels of alanine, aspartic acid, valine, linoleic acid, linolenic acid, oleic acid, plamitic acid, stearic acid, glucosylglycerol, sucrose, phosphatidylglycerol (PG), and diacylglyceryltrimethylhomoserine (DGTS) (only detected in Synechocystis 7338) were significantly higher in Synechocystis 7338. Our results provide basic information to choose Synechocystis strains for various industrial usages, and also provide fundamental information on the future development of novel Synechocystis strains to produce high levels of specific metabolites by metabolic pathway modulation.

Microalgal biomass productivity of semi-permeable membrane photobioreactors (SPM-PBRs) highly depends on diffusion of nutrient ions from surrounding environment through SPMs. The objective of this study was to enhance microalgal biomass productivity in floating photobioreactors (PBRs) with semi-permeable membranes (SPMs) grafted with 4-hydroxyphenethyl bromide (4-HPB) by increasing ion permeabilities and reducing biofouling of SPMs. In this study, we investigated the reduction of biofouling and improvement of biomass productivity by grafting 4-HPB onto SPMs which could decrease hydrophobicity. The degree of alkylation could be increased by up to 4.9%, with water contact angles on alkylated SPM surfaces decreased from 102.5° to 58.2°, indicating decreased surface hydrophobicity. Ion and water permeabilities of SPMs were increased in alkylated SPMs by 148% and 200%, respectively, compared to non-treated SPM control. Moreover, biomass productivity of PBRs with alkylated SPMs was 75% (740 mg m−2 day−1) higher than that of the control. Biofouling was also remarkably reduced by up to 40% in 4-HPB grafted SPMs after cultivation of microalgae. These results suggest that decreasing hydrophobicity of SPMs by grafting 4-HPB could enhance biomass productivity in SPM-PBRs through improving ion permeability and reducing biofouling.

Environmentally benign silver nanoparticles (Ag NPs) were synthesized at different temperatures using leaf extract of Platycodon grandiflorum. The apparent UV–Vis spectrum maxima clearly displayed plasmon resonance bands at 442 and 457 nm for nanoparticles synthesized at 37 °C and 50 °C, respectively. The morphology of Ag NPs were investigated by FESEM, FETEM and AFM analysis and the results showed the formation of uniform spherical nanoparticles with average sizes of 19 and 21 nm for Ag NPs produced at 37 °C and 50 °C, respectively, with slightly larger nanoparticles formed at 50 °C owing to agglomeration. Further characterization by XRD, XPS, and FTIR confirmed their crystal nature, while stability was confirmed by zeta potential analysis. The strong XRD diffraction pattern confirmed the face-central cubic crystalline nature of the nanoparticles. Moreover, XPS revealed Ag 3d doublet (3d5/2 and 3d3/2) with C1s peaks of the catalysts that were turned into C H (283.68) C C, C C (284.16 eV), C O (285.34 eV), and C O (288.55 eV). The O1s spectra were also exhibited with binding energy at 530.94, 531.96 and 532.53 for AgO, C O and C O, respectively, and FTIR confirmed the peaks of plant extract appeared on Ag NPs. Overall, Ag NPs synthesized at 50 °C had good shape and structure with a high stability (−5.23). The nanoparticles also exhibited antibacterial activity against Escherichia coli and Bacillus subtilis. The biosynthesized Ag NPs have the potential for use in various applications, particularly in the field of biomedicine.

Nanoparticles have been paid a great attraction in interdisciplinary sciences owing to their unique characteristics. Different nanoparticles were synthesized in metallic and non-metallic states, showing varied sizes for down-stream applications. Copper nanoparticle (CuNP) has a special attention due to its great electrical, catalytic, optical and mechanical properties. In this research, CuNP was prepared by the electrolytic deposition and characterized its morphological and structural behaviours by scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), X-ray powder diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and Raman spectroscopy. SEM displays the irregular agglomerated particles, whereas with TEM the sizes were determined as 40 ± 10 nm. The peaks with XPS analysis were noticed at 932.1 for Cu(2p3/2), 944.1 eV and 952.1 eV for Cu(2p1/2). Whereas, the representative peaks for copper were placed at 43.7°, 50.7° and 74.3°, corresponding to (111), (200) and (220) under XRD observations. CuNP vibration peaks were monitored at 1635 cm−1 with FTIR and a sharp peak at 617.6 cm−1 is representing Cu-O group due to the occurrence of Cu2O. The antimicrobial activity of the electrodeposited CuNP was proved against the fungus, Aspergillus niger. Further, ITO/CuNP/CuI/Cu (Indium Tin Oxide/Copper Nanoparticle/Copper Iodide/Copper) substrate solid-state photovoltaic cell was fabricated by CuNP electrodeposition on the CuI substrate. It was anchored by I–V characteristics, apparent that a rapid current enhancement after deposition of the CuNP.

The rapid and reproducible preparation of biological samples is an essential requirement for LC-MS-based clinical metabolomic profiling. Herein, we proposed a novel, automated sample preparation method for serum metabolite LC-MS analysis. This method included a one-step double-filtration process with two filters (30k and 10k MWCO). High sample throughput (up to 96 samples) was achieved in an automated manner by incorporating the process in a liquid handling robotic system. The effectiveness of the developed method was evaluated through comparison with a conventional manual centrifugation-based method using seven human serum samples. A higher metabolite recovery and better reproducibility were achieved in quantitative analysis using the automated double-filtration method. The results suggested that the multiplexed serum sample preparation system can be applied to large-scale untargeted metabolomics studies.

There is increasing demand for essential fatty acids (EFAs) from non-fish sources such as microalgae, which are considered a renewable and sustainable biomass. The open raceway system (ORS) is an affordable system for microalgae biomass cultivation for industrial applications. However, seasonal variations in weather can affect biomass productivity and the quality of microalgal biomass. The aim of this study was to determine the feasibility of year-round Tetraselmis sp. cultivation in a semi-ORS in Korea for biomass and bioactive lipid production. To maximize biomass productivity of Tetraselmis sp., f medium was selected because it resulted in a significantly higher biomass productivity (1.64 ± 0.03 g/L) and lower omega-6/omega-3 ratio (0.52/1) under laboratory conditions than f/2 medium (0.70/1). Then, we used climatic data-based building information modeling technology to construct a pilot plant of six semi-ORSs for controlling culture conditions, each with a culture volume of 40,000 L. Over 1 year, there were no significant variations in monthly biomass productivity, fatty acid composition, or the omega-6/omega-3 ratio, however, the lipid content correlated significantly with photosynthetic photon flux density. During year-round cultivation from November 2014 to October 2017, areal productivity was gradually increased by increasing medium salinity and injecting CO2 gas into the culture medium. Productivity peaked at 44.01 g/m2/d in October 2017. Throughout the trials, there were no significant differences in average lipid content, which was 14.88 ± 1.26%, 14.73 ± 2.44%, 12.81 ± 2.82%, and 13.63 ± 3.42% in 2014, 2015, 2016, and 2017, respectively. Our results demonstrated that high biomass productivity and constant lipid content can be sustainably maintained under Korean climate conditions.

Synechocystis strains are cyanobacteria that can produce useful biomaterials for biofuel and pharmaceutical resources. In this study, the effects of exogenous glucose (5-mM) on cell growth, photosynthetic pigments, metabolites, and lipids in Synechocystis sp. PCC 7338 (referred to as Synechocystis 7338) were investigated. Exogenous glucose increased cell growth on days 9 and 18. The highest production (mg/L) of chlorophyll a (34.66), phycocyanin (84.94), allophycocyanin (34.28), and phycoerythrin (6.90) was observed on day 18 in Synechocystis 7338 culture under 5-mM glucose. Alterations in metabolic and lipidomic profiles under 5-mM glucose were investigated using gas chromatography-mass spectrometry (MS) and nanoelectrospray ionization-MS. The highest production (relative intensity/L) of aspartic acid, glutamic acid, glycerol-3-phosphate, linolenic acid, monogalactosyldiacylglycerol (MGDG) 16:0/18:1, MGDG 16:0/20:2, MGDG 18:1/18:2, neophytadiene, oleic acid, phosphatidylglycerol (PG) 16:0/16:0, and PG 16:0/17:2 was achieved on day 9. The highest production of pyroglutamic acid and sucrose was observed on day 18. We suggest that the addition of exogenous glucose to Synechocystis 7338 culture could be an efficient strategy for improving growth of cells and production of photosynthetic pigments, metabolites, and intact lipid species for industrial applications.

Biodiesel contains methyl or ethyl esters of long-chain fatty acids and has recently attracted increasing attention. Microalgae have emerged as a sustainable biodiesel production system owing to their photosynthetic potential. However, the conversion of microalgal biomass to biodiesel requires high energy and is costly. This study aimed to overcome the high cost of the pretreatment process by generating cyanobacteria converting fatty acids to fatty acids methyl ester (FAME) in vivo by introducing the fatty acid methyl ester transferase (FAMT) gene. Two FAMT genes from Drosophila melanogaster and Arabidopsis thaliana were selected and their codons were optimized for insertion in the Synechocystis sp. PCC6803 genome through homologous recombination, respectively. FAMT mRNA and protein expression levels were confirmed through reverse-transcription PCR and western blot analysis, respectively. Furthermore, heterologous expression of the FAMT genes yielded FAME, which was analyzed by gas chromatography. We found that FAMT transformants can be further metabolically optimized and applied for commercial production of biodiesel.

Cyanobacteria, given their ability to produce various secondary metabolites utilizing solar energy and carbon dioxide, are a potential platform for sustainable production of biochemicals. Until now, conventional metabolic engineering approaches have been applied to various cyanobacterial species for enhanced production of industrially valued compounds, including secondary metabolites and non-natural biochemicals. However, the shortage of understanding of cyanobacterial metabolic and regulatory networks for atmospheric carbon fixation to biochemical production and the lack of available engineering tools limit the potential of cyanobacteria for industrial applications. Recently, to overcome the limitations, synthetic biology tools and systems biology approaches such as genome-scale modeling based on diverse omics data have been applied to cyanobacteria. This review covers the synthetic and systems biology approaches for advanced metabolic engineering of cyanobacteria.

Freshwater Synechocystis sp. PCC 6803 has been considered to be a platform for the production of the next generation of biofuels and is used as a model organism in various fields. Various genomics, transcriptomics, metabolomics, and proteomics studies have been performed on this strain, whereas marine Synechocystis sp. PCC 7338 has not been widely studied despite its wide distribution. This study analyzed the proteome profiles of two Synechocystis strains using a liquid chromatography–tandem mass spectrometry-based bottom-up proteomic approach. Proteomic profiling of Synechocystis sp. PCC 7338 was performed for the first time with a data-dependent acquisition method, revealing 18,779 unique peptides and 1794 protein groups. A data-independent acquisition method was carried out for the comparative quantitation of Synechocystis sp. PCC 6803 and 7338. Among 2049 quantified proteins, 185 up- and 211 down-regulated proteins were defined in Synechocystis sp. PCC 7338. Some characteristics in the proteome of Synechocystis sp. PCC 7338 were revealed, such as its adaptation to living conditions, including the down-regulation of some photosynthesis proteins, the up-regulation of kdpB, and the use of osmolyte glycine as a substrate in C1 metabolism for the regulation of carbon flow. This study will facilitate further studies on Synechocystis 7338 to define in depth the proteomic differences between it and other Synechocystis strains.

Hydrothermal liquefaction of Chlorella vulgaris feedstock containing 80% (w/w) water was conducted in a batch reactor as a function of temperature (300, 325 and 350 °C) and reaction times (5, 10 and 30 min). The biocrude yield, elemental composition and higher heating value obtained for various reaction conditions helped to predict the optimum conditions for maximizing energy recovery. To optimize the recovery of inorganic nutrients, we further investigated the effect of reaction conditions on the ammonium (NH4+), phosphate (PO43−), nitrate (NO3−) and nitrite (NO2−) concentrations in the aqueous phase. A maximum energy recovery of 78% was obtained at 350 °C and 5 min, with a high energy density of 34.3 MJ/kg and lower contents of oxygen. For the recovery of inorganic nutrients, shorter reaction times achieved higher phosphorus recovery, with maximum recovery being 53% at 350 °C and 5 min. Our results indicate that the reaction condition of 350 °C for 5 min was optimal for maximizing energy recovery with improved quality, at the same time achieving a high phosphorus recovery.

Novel carbon-based solid acid catalysts were synthesized through a sustainable route from lipid-extracted microalgal residue of Dunaliella tertiolecta, for biodiesel production. Two carbon-based solid acid catalysts were prepared by surface modification of bio-char with sulfuric acid (H₂SO₄) and sulfuryl chloride (SO₂Cl₂), respectively. The treated catalysts were characterized and their catalytic activities were evaluated by esterification of oleic acid. The esterification catalytic activity of the SO₂Cl₂-treated bio-char was higher (11.5 mmol Prod.∙h⁻¹∙g Cat. ⁻¹) than that of commercial catalyst silica-supported Nafion SAC-13 (2.3 mmol Prod.∙h⁻¹∙g Cat. ⁻¹) and H₂SO₄-treated bio-char (5.7 mmol Prod.∙h⁻¹∙g Cat. ⁻¹). Reusability of the catalysts was examined. The catalytic activity of the SO₂Cl₂-modified catalyst was sustained from the second run after the initial activity dropped after the first run and kept the same activity until the fifth run. It was higher than that of first-used Nafion. These experimental results demonstrate that catalysts from lipid-extracted algae have great potential for the economic and environment-friendly production of biodiesel.

This study focused on the evaluation of antimicrobial activity of silver nanoparticles (AgNPs) after their green synthesis by means of a Solanum trilobatum bark extract. The obtained product with an intense surface plasmon resonance band at ∼442 nm with UV–visible spectroscopic analysis indicated the formation of AgNPs. The morphology of AgNPs was observed under transmission electron microscopy and field emission scanning electron microscopy, displayed that the eco-friendly synthesized AgNPs have a spherical shape with an average size of ∼25 nm in diameter. X-ray powder diffraction and selected area electron diffraction analyses confirmed that the AgNPs are crystalline in nature. Fourier transform infrared spectroscopy indicated that the AgNPs capped with active ingredients of the bark extract. X-ray photoelectron spectroscopy revealed elemental composition of the AgNPs. The performance of S. trilobatum bark extract-capped AgNPs in terms of inhibition of microbial growth was studied by disc diffusion and well diffusion assays. Eco-friendly synthesized S. trilobatum extract-capped AgNPs were found to possess enhanced antimicrobial properties: growth inhibition of gram-negative and gram-positive bacteria and of fungal species. These results demonstrated the potential applications of the indigenous medicinal plants to the field of nanotechnology.

Lipids in microalgae are energy-rich compounds and considered as an attractive feedstock for biodiesel production. To redirect carbon flux from competing pathways to the fatty acid synthesis pathway of Tetraselmis sp., we used three types of chemical inhibitors that can block the starch synthesis pathway or photorespiration, under nitrogen-sufficient and nitrogen-deficient conditions. The starch synthesis pathway in chloroplasts and the cytosol can be inhibited by 3-(3,4-dichlorophenyl)-1,1-dimethylurea and 1,2-cyclohexane diamine tetraacetic acid (CDTA), respectively. Degradation of glycine into ammonia during photorespiration was blocked by aminooxyacetate (AOA) to maintain biomass concentration. Inhibition of starch synthesis pathways in the cytosol by CDTA increased fatty acid productivity by 27% under nitrogen deficiency, whereas the blocking of photorespiration in mitochondria by AOA was increased by 35% under nitrogen-sufficient conditions. The results of this study indicate that blocking starch or photorespiration pathways may redirect the carbon flux to fatty acid synthesis.

Lipid extraction is a critical step in the downstream processing of biodiesel production from microalgae. Solvent extraction using mixtures of non-polar and polar solvents is one of the most well-known processes for this purpose. Hexane is the most common solvent of choice for large-scale lipid extractions due to its technical and economic advantages, especially its high selectivity toward lipids and low cost. In this study, extractions using mixtures of hexane and polar solvents were evaluated for their performance in order to develop a more efficient method for large-scale lipid extraction from microalgae. The combination of hexane and methanol resulted in the highest fatty acid methyl ester (FAME) yield for lipids from Tetraselmis sp. The effects of extraction conditions, including proportions of methanol to hexane, ratios of total solvent volume to dry biomass, and extraction time, on extraction yields were evaluated to determine optimum conditions providing higher lipid and FAME yields. The optimal conditions were as follows: proportion of hexane to methanol of 1:1, ratio of total solvent volume to dry biomass of 10 mL/g, and extraction time of 120 min. Finally, the selected solvent mixture and optimal conditions were applied to larger scale extraction experiments with scale-up factors of 10, 50, and 100. FAME yields of large-scale extractions were almost completely consistent with increasing scale-up factors. The results of this study suggest that a hexane and methanol mixture is a promising solvent for large-scale lipid extraction from microalgae.

In this study, hypersaline media were used for ocean cultivation of the marine microalga Tetraselmis sp. KCTC12432BP for enhanced biomass and fatty acid (FA) productivity. Hypersaline media (55, 80, and 105 PSU) were prepared without sterilization by addition of NaCl to seawater obtained from Incheon, Korea. The highest biomass productivity was obtained at 55 PSU (0.16 g L−1 day−1) followed by 80 PSU (0.15 g L−1 day−1). Although the specific growth rate of Tetraselmis decreased at salinities higher than 55 PSU, prevention of contamination led to higher biomass productivity at 80 PSU than at 30 PSU (0.03 g L−1 day−1). FA content of algal biomass increased as salinity increased to 80 PSU, above which it declined, and FA productivity was highest at 80 PSU. Ocean cultivation of Tetraselmis was performed using 50-L tubular module photobioreactors and 2.5-kL square basic ponds, closed- and open-type ocean culture systems, respectively. Culturing microalgae in hypersaline medium (80 PSU) improved biomass productivities by 89 and 152% in closed and open cultures, respectively, compared with cultures with regular salinity. FA productivity was greatly improved by 369% in the closed cultures. The efficacy of salinity shift and N-deficiency to enhance FA productivity was also investigated. Lowering salinity to 30 PSU with N-starvation following cultivation at 80 PSU improved FA productivity by 19% in comparison with single-stage culture without N-deficiency at 30 PSU. The results show that salinity manipulation could be an effective strategy to improve biomass and FA productivity in ocean cultivation of Tetraselmis sp.

Hydrothermal liquefaction (HTL) is a thermal depolymerization process used to convert wet biomass such as microalgae into bio-crude oil. Four main products, gas, bio-crude oil, an aqueous phase, and solid residue, are generated through HTL. In this study, various HTL conditions were investigated to enhance the phosphorus content in the aqueous phase as well as the yield of bio-crude oil. Tetraselmis sp. was used as the microalgae feedstock, and the product yields according to catalyst type were explored. The phosphate ion (PO43−) content in the aqueous phase was significantly enhanced when acetic acid was added because of the effect of pH. In addition, it was found that both the bio-crude oil yield and the ammonium ion content could be increased by recycling the post-HTL aqueous phase, while the phosphate content was not.

Some artificial light sources able to emit photons at specific wavelengths, such as LEDs, are useful for studying the effects of light quality on microalgal growth and production of fatty acids; however, they should not be used for outdoor cultivation of microalgae to produce bioenergy. Instead, various light filters capable of selectively transmitting red, blue, and red+blue light regions in solar radiation were used to cover 0.4 L bubble column photobioreactors to cultivate Tetraselmis sp. KCTC12236BP and investigate the influence of light quality on microalgal growth and fatty acid production. Biomass and fatty acid productivities in red light (0.10 ± 0.05 g/L/day and 11.8 ± 0.5 mg/L/day, respectively) were 7 ~ 53% and 9 ~ 61% higher than other colored lights based on the same number of supplied photons, respectively. The composition of fatty acids did not change significantly in response to transmitted light qualities of the filter. The ratio of saturated to unsaturated fatty acids was 3:7, and their contents were 12% in all groups, which corresponds with the results of LEDs. Plotting biomass and fatty acid productivity over the red photon fraction in supplied light revealed that increased productivities were closely correlated with red photon fraction in the filtered light. Overall, the results presented herein indicate that enhanced production of algal fatty acid could be achieved by application of light filters in outdoor settings without artificial lights.

Microalgae are attracting much attention as superior biodiesel producers. In particular, under stressful conditions, they accumulate organic compounds consisting entirely of carbon and hydrogen. The aim of this work was to increase intracellular fatty acid content in Dunaliella tertiolecta (Chlorophyceae), Nannochloropsis oculata (Eustigmatophyceae), and Porphyridium cruentum (Rhodophyceae) using a combination of nitrogen starvation and chemical inhibitors of carbohydrate biosynthesis. These microalgae were subjected to nitrogen starvation and their physiological changes were then observed over time. In D. tertiolecta, no significant change in total fatty acid content was detected on day 3.5 relative to the initial total fatty acid content (day 0), while total carbohydrate content dramatically increased as the nitrogen starvation period was extended. In N. oculata, total fatty acid content rapidly increased, reaching up to nearly 40% of the DCW at day 3.5. However, total carbohydrate content exhibited a gradual reduction throughout the experiment. In P. cruentum, total carbohydrate content increased up to 43% of DCW on day 3.5 and total fatty acid content increased slightly under nitrogen depletion. These data suggest that different eukaryotic microalgae use different storage products under stressful conditions. Among the three strains, D. tertiolecta showed decreased total carbohydrate content and enhanced total fatty acid content following inhibition of carbohydrate synthesis by dichlorophenyl dimethylurea and cyclohexane diamine tetra acetic acid. The results demonstrate the possibility of furthering our understanding of the fatty acid and carbohydrate biosynthesis metabolic network that responds to environmental changes in microalgae.

Microalgae and bacteria are known to be closely associated in diverse environments. To isolate dominant bacterial species associated with a green alga, Dunaliella tertiolecta, a photoreactor culture of the microalga was investigated using culture-based and culture-independent approaches. The bacterial community structure of the algal culture showed that the most abundant bacterial species under the culture conditions was related to the genus Winogradskyella. The closely related amplicon sequences, showing ≥ 99.5% 16S rRNA gene sequence similarity to one of the isolates, designated IMCC-33238T, constituted > 49% of the bacterial community and was therefore regarded as the most dominant species in the algal culture. Strain IMCC33238T was characterized by Gramstaining-negative and orange-colored rods. Phylogenetic analyses of the 16S rRNA genes as well as whole genome sequences revealed that strain IMCC33238T belonged to Winogradskyella and shared more than 97.2% 16S rRNA gene sequence similarity with Winogradskyella species. The strain contained iso-C15:1 G, iso-C15:0, iso-C15:0 3-OH, and summed feature 3 (C16:1ω6c and/or C16:1ω7c) as major fatty acids and MK-6 as the predominant quinone. The polar lipids found in strain IMCC33238T were phosphatidylethanolamine, two unidentified aminolipids, and three unidentified lipids. The genome of strain IMCC33238T was 3.37 Mbp in size with 33.9 mol% G + C content and proteorhodopsin. Many genes encoding folate and vitamin production are considered to play an important role in the bacteria-algae interaction. On the basis of phylogenetic and phenotypic characteristics, strain IMCC33238T represents a novel species in the genus Winogradskyella, for which the name Winogradskyella algicola sp. nov. is proposed. The type strain is IMCC33238T (= KACC 21192T = NBRC 113704T).

Chemically-processed clays (PRCs) by base and acid treatments were developed to effectively remove microalgae, and their removal efficiencies were evaluated and compared to that of natural clay. The processed clays were produced by using sodium hydroxide and different amounts of sulfuric acid, and their characteristics were analyzed. Microalgae removal efficiency of the clay was varied depending on the amount of sulfuric acid used in treatments. When adding 1 g/L of a PRC-D type (treated with 1.5 v/w sulfuric acid) to the Microcystis aeruginosa suspension (1 × 106 cells/mL), the highest removal performance (87.3 ± 1.5%) without significant pH variation of the suspension was found among tested PRCs, whereas that of natural clay was 32.1 ± 2.2%. Finally, when selected PRC-D showing best efficiency was applied to lake water taken at a local algae bloomed lake, it exhibited 77.6% removal efficiency of microalgae, indicating 1.7 times higher than the natural clay. The results suggest that processed clays in this study could contribute to effective removal of microalgae in the algal-bloomed area with minimal environmental impact.

Microalgae have long been considered as one of most promising feedstocks with better characteristics for biofuels production over conventional energy crops. There have been a wide range of estimations on the feasibility of microalgal biofuels based on various productivity assumptions and data from different scales. The theoretical maximum algal biofuel productivity, however, can be calculated by the amount of solar irradiance and photosynthetic efficiency (PE), assuming other conditions are within the optimal range. Using the actual surface solar irradiance data around the world and PE of algal culture systems, maximum algal biomass and biofuel productivities were calculated, and feasibility of algal biofuel were assessed with the estimation. The results revealed that biofuel production would not easily meet the economic break‐even point and may not be sustainable at a large‐scale with the current algal biotechnology. Substantial reductions in the production cost, improvements in lipid productivity, recycling of resources, and utilization of non‐conventional resources will be necessary for feasible mass production of algal biofuel. Among the emerging technologies, cultivation of microalgae in the ocean shows great potentials to meet the resource requirements and economic feasibility in algal biofuel production by utilizing various marine resources.

A split-column photobioreactor (SC-PBR), consisting of two bubble columns with different sizes, was developed to enhance the photon utilization efficiency in an astaxanthin production process from Haematococcus lacustris. Among the two columns, only the smaller column of SC-PBR was illuminated. Astaxanthin productivities and photon efficiencies of the SC-PBRs were compared with a standard bubble-column PBR (BC-PBR). Astaxanthin productivity of SC-PBR was improved by 28%, and the photon utilization efficiencies were 28-366% higher than the original BC-PBR. The results clearly show that the effective light regime of SC-PBR could enhance the production of astaxanthin.

In this study, Chlorella vulgaris (C. vulgaris) was treated with ethephon at low (50 μM) and high (200 μM) concentrations in medium and harvested at 0, 7, and 14 days, respectively. The presence of ethephon led to significant metabolic changes in C. vulgaris, with significantly higher levels of α-tocopherol, γ-aminobutyric acid (GABA), asparagine, and proline, but lower levels of glycine, citrate, and galactose relative to control. Ethephon induced increases in saturated fatty acids but decreases in unsaturated fatty acids. The levels of highly saturated sulfoquinovosyldiacylglycerol species and palmitic acid bound phospholipids were increased on day 7 of ethephon treatment. Among the metabolites, the productivities of α-tocopherol (0.70 μg/L/day) and GABA (1.90 μg/L/day) were highest for 50 and 200 μM ethephon on day 7, respectively. We propose that ethephon treatment involves various metabolic processes in C. vulgaris and can be an efficient way to enrich the contents of α-tocopherol and GABA.

This study investigated the effects of different temperature conditions on the comprehensive metabolite profiles of Chlorella vulgaris using gas chromatography–mass spectrometry coupled with multivariate statistical analysis. C. vulgaris cells cultivated at 20 °C were transferred to 10 °C incubators at different time points of cultivation [days 0 (TR0D), 7 (TR7D), and 14 (TR14D)], then they were cultivated at 10 °C until harvesting at day 21 to compare the growth and comprehensive metabolite profiles with those cultivated under a constant cultivation temperature of 20 °C (T20). There was no significant difference in algal cell growth between cultivation under the T20 and temperature reduction (TR) conditions. Algal fatty-acid profiles under TR were different from those of the T20 condition. Specifically, the contents of octadecanoic acid (C18:0), octadecenoic acid (C18:1), hexadecadienoic acid (C16:2), and octadecadienoic acid (C18:2) increased the most under TR0D. The relative levels of metabolites such as β-alanine, glutamine, glycine, isoleucine, proline, valine, and myo-inositol, which act as osmolytes, and bioactive compounds such as neophytadiene and ascorbic acid were increased under TR conditions on day 21. Among the metabolites, the contents of neophytadiene and ascorbic acid were further investigated, and the content of ascorbic acid was highest on day 14 under the TR7D condition, while the content of neophytadiene was highest on day 21 under the TR0D and TR14D condition. Therefore, we suggest that a TR from 20 to 10 °C could enhance the production in C. vulgaris cultures of bioactive fatty acids such as C18:1, C16:2, and C18:2 (TR0D), organic osmolytes such as β-alanine, glutamine, glycine, isoleucine, proline, valine, and myo-inositol (TR conditions), ascorbic acid (TR7D), and neophytadiene (TR0D and TR14D).

Lumostatic operation was applied for efficient astaxanthin production in autotrophic Haematococcus lacustris cultures using 0.4-L bubble column photobioreactors. The lumostatic operation in this study was performed with three different specific light uptake rates (q e) based on cell concentration, cell projection area, and fresh weight as one-, two- and three-dimensional characteristics values, respectively. The q e value from the cell concentration (q e1D) obtained was 13.5 × 10−8 μE cell−1 s−1, and the maximum astaxanthin concentration was increased to 150 % compared to that of a control with constant light intensity. The other optimum q e values by cell projection area (q e2D) and fresh weight (q e3D) were determined to be 195 μE m−2 s−1 and 10.5 μE g−1 s−1 for astaxanthin production, respectively. The maximum astaxanthin production from the lumostatic cultures using the parameters controlled by cell projection area (2D) and fresh weight (3D) also increased by 36 and 22 % over that of the controls, respectively. When comparing the optimal q e values among the three different types, the lumostatic cultures using q e based on fresh weight showed the highest astaxanthin productivity (22.8 mg L−1 day−1), which was a higher level than previously reported. The lumostatic operations reported here demonstrated that more efficient and effective astaxanthin production was obtained by H. lacustris than providing a constant light intensity, regardless of which parameter is used to calculate the specific light uptake rate.

Culturing microalgae in the ocean has potentials that may reduce the production cost and provide an option for an economic biofuel production from microalgae. The ocean holds great potentials for mass microalgal cultivation with its high specific heat, mixing energy from waves, and large cultivable area. Suitable photobioreactors (PBRs) that are capable of integrating marine energy into the culture systems need to be developed for the successful ocean cultivation. In this study, prototype floating PBRs were designed and constructed using transparent low-density polyethylene film for microalgal culture in the ocean. To improve the mixing efficiency, various types of internal partitions were introduced within PBRs. Three different types of internal partitions were evaluated for their effects on the mixing efficiency in terms of mass transfer (k(L)a) and mixing time in the PBRs. The partition type with the best mixing efficiency was selected, and the number of partitions was varied from one to three for investigation of its effect on mixing efficiency. When the number of partitions is increased, mass transfer increased in proportion to the number of partitions. However, mixing time was not directly related to the number of partitions. When a green microalga, Tetraselmis sp. was cultivated using PBRs with the selected partition under semi-continuous mode in the ocean, biomass and fatty acid productivities in the PBRs were increased by up to 50 % and 44% at high initial cell density, respectively, compared to non-partitioned ones. The results of internally partitioned PBRs demonstrated potentials for culturing microalgae by efficiently utilizing ocean wave energy into culture mixing in the ocean.

A marine green microalga, Tetraselmis sp., has been studied for the production of biomass and lipids in seawater culture. Since carbohydrate and lipid biosynthesis are competitive metabolic pathways, we attempted to increase lipid synthesis in Tetraselmis by inhibiting carbohydrate synthesis. The main regulatory enzyme in the starch synthesis pathway is ADP-glucose pyrophosphorylase (AGP). AGP loss-of-function mutants were developed using the CRISPR-Cas9 ribonucleoprotein (RNP) delivery system. AGP mutants showed a slight decrease in growth. However, the lipid content in two AGP mutants was significantly enhanced by 2.7 and 3.1 fold (21.1% and 24.1% of DCW), respectively, compared to that in the wild type (7.68% of DCW) under nitrogen starvation. This study is an example of metabolic engineering by genetic editing using the CRISPR-Cas9 RNP method in marine green microalgae. Consequently, starchless Tetraselmis mutants might be considered potential producers of lipids in seawater cultures.

The Chinese traditional joss paper has been recycled and silica and aluminium nanocomposite production is reported in the current research, using an alkalinization procedure. Characterization of silica and aluminium nanocomposite extracted was performed using field-emission transmission electron microscope (FETEM), field-emission scanning electron microscope (FESEM), UV–visible spectroscopy, high-power optical microscopy (HPM) and 3D nano profiler, indicates the size, shape and particle distribution, eventually reveals the purity of silica and aluminium nanocomposite synthesized. The silica-aluminium nanocrystal synthesized from joss paper waste was found to be ~ 15 nm size range under FETEM analysis and FESEM analysis revealed the uniform spherical shape. The energy disperse spectroscopy (EDX) data attained from FESEM analysis affirmed the synthesis of silica-aluminium nanocomposite with greater purity since the ratio of silicon to aluminium observed in EDX spectra was 13.24 to 7.96. HPM indicated the size of agglomerated aluminosilicate is 36.74 μm whereas 3D nano profiler has proven that the height of nanocrystal synthesized from joss paper waste is 234.37 μm. Band gap value of 3.84 eV, calculated using UV–vis absorbance readings further affirmed the size of nanocomposite which is in good agreement with presented results.

This study aimed to improve the production of phycobiliproteins using TiO2 nanoparticles (NPs) in Synechocystis sp. PCC 6803. The growth characteristics of Synechocystis cells were not affected by TiO2 NPs treatment, but this treatment increased the chlorophyll content significantly by 62.2% (14.6 mg/L) compared to that of control (9.0 mg/L) on day 16. Phycocyanin production was increased by 33.8% (29.3 g/L) compared to that of control (21.9 g/L) on day 8. Allophycocyanin production was increased by 55.0% (6.2 g/L) compared to that of control (4.0 g/L) on day 8, and by 22.4% (16.4 g/L) compared to that of control (13.4 g/L) on day 16. Direct infusion mass spectrometry revealed that TiO2 NPs treatment significantly increased the levels of major thylakoid membranes of monogalactosyldiacylglycerols (18:2/18:3, 18:2/18:2, 18:1/18:2), phosphatidylglycerol (16:0/16:1), and sulfoquinovosyldiacylglycerols (16:0/16:1, 16:0:18:4) on day 8. These findings indicate that TiO2 NPs have potential for commercial applications in Synechocystis species or other microalgal strains.

Microalgae are promising candidates for biofuel production due to their high lipid content. To facilitate utilization of the microalgae for biofuel, rapid quantification of the lipid contents in microalgae is necessary. However, conventional methods based on the chemical extraction of lipids require a time-consuming destructive extraction process. Here, we demonstrate label-free, non-invasive, rapid quantification of the lipid contents in individual micro-algal cells measuring the three-dimensional refractive index tomograms. We measure three-dimensional refractive index distributions within Nannochloropsis oculata cells and find that lipid droplets are identifiable in tomograms by their high refractive index. In addition, we alter N. oculata under nitrogen deficiency by measuring the volume, lipid weight, and dry cell weight of individual cells. Characterization of individual cells allows correlative analysis between the lipid content and size of individual cells.