Purification and Screening of the Antialgal Activity of Seaweed Extracts and a New Glycolipid Derivative against Two Ichthyotoxic Red Tide Microalgae Amphidinium carterae and Karenia mikimotoi.

Ichthyotoxic red tide is a problem that the world is facing and needs to solve. The use of antialgal compounds from marine macroalgae to suppress ichthyotoxic red tide is considered a promising biological control method. Antialgal substances were screened and isolated from Bangia fusco-purpurea, Gelidium amansii, Gloiopeltis furcate, Hizikia fusifarme, Laminaria japonica, Palmaria palmata, and Sargassum sp. to obtain new materials for the development of algaecides against ichthyotoxic red tide microalgae using bioactivity-guided isolation methods. The fractions of seven macroalgae exhibited selective inhibitory activities against Amphidinium carterae and Karenia mikimotoi, of which the ethyl acetate fractions had the strongest and broadest antialgal activities for the two tested red tide microalgae. Their inhibitory effects on A. carterae and K. mikimotoi were even stronger than that of potassium dichromate, such as ethyl acetate fractions of B. purpurea, H. fusifarme, and Sargassum sp. Thin-layer chromatography and ultraviolet spectroscopy were further carried out to screen the ethyl acetate fraction of Sargassum sp. Finally, a new glycolipid derivative, 2-O-eicosanoyl-3-O-(6-amino-6-deoxy)-ß-D-glucopyranosyl-glycerol, was isolated and identified from Sargassum sp., and it was isolated for the first time from marine macroalgae. The significant antialgal effects of 2-O-eicosanoyl-3-O-(6-amino-6-deoxy)-ß-D-glucopyranosyl-glycerol on A. carterae and K. mikimotoi were determined.

Effects of culture temperature and light regimes on biomass and lipid accumulation of Chlamydomonas reinhardtii under carbon-rich and nitrogen-limited conditions.

This study investigated the impacts of various culture temperatures and light regimes on growth and biochemical constituents of Chlamydomonas reinhardtii under carbon-supply and nitrogen-limited conditions to improve oil production in algal cells. Results displayed that under a 30 ℃ and 150 µE/m2/s regime, there was a significant increase in biomass, total lipids, and lipid productivity. Specifically, these parameters reached 1.83 g/L, 36.25 %, and 130.73 mg/L/d, respectively. Remarkably, prolonging the photoperiod further enhanced the aforementioned three parameters, reaching peak levels of 1.92 g/L, 41.10 %, and 157.54 mg/L/d, respectively, recorded at a 24/0h photoperiod. Compared with cultures grown under normal conditions, these values displayed increments of 1.21-fold, 74.88 %, and 3.01-fold, respectively. Additionally, under optimal conditions, the soluble sugar content reached 79.72 mg/g, and the biodiesel properties were improved. These findings indicate that moderately increasing temperature, light intensity, and photoperiod could achieve the co-production of biomass, lipids, and sugars in C. reinhardtii.

Purification efficiency of Pyropia-processing wastewater and microalgal biomass production by the combination of Chlorella sp. C2 cultivated at different culture temperatures and chitosan.

To elucidate the impacts of culture temperature on nutrient removal efficiency of Pyropia-processing wastewater (PPW) and microalgal biomass production, Chlorella sp. C2 was employed and cultivated in raw PPW under different temperatures. Results showed that, after incubating for 7 days, higher biomass (0.50 g/L) and total lipids (21.84 %) were attained at 35 °C. The maximal chemical oxygen demand (COD), phycobiliprotein, total nitrogen and total phosphorus removal rates were observed at 30-35 °C and separately reached 62.41 %, 92.61 %, 92.19 % and 98.33 %. Interestingly, COD removal efficiencies of Chlorella cells, cultivated for 3, 5 and 7 days at 30-35 °C, 15-25 °C and 10 °C respectively, could reach >75 % with assistance from 60-80 mg/L chitosan. Meanwhile, the clarification efficiency of chitosan on algal cells reached >95 %. It suggests that Chlorella strain cultured at altered temperatures could efficiently remove PPW nutrients assisted by moderate chitosan, simultaneously achieving the rapid harvest of microalgae.

Reducing culture medium nitrogen supply coupled with replenishing carbon nutrient simultaneously enhances the biomass and lipid production of Chlamydomonas reinhardtii.

Chlamydomonas reinhardtii is a model strain to explore algal lipid metabolism mechanism, and exhibits great potentials in large-scale production of lipids. Completely lacking nitrogen is an efficient strategy to trigger the lipid synthesis in microalgal cells, while it always leads to the obvious reduction in the biomass. To illustrate the optimal culture substrate carbon (C) and nitrogen (N) levels to simultaneously stimulate the growth and lipid production of C. reinhardtii, cells were cultivated under altered C and N concentrations. Results showed that replenishing 6 g/L sodium acetate (NaAc) could increase 1.50 and 1.53 times biomass and lipid productivity compared with 0 g/L NaAc treatment (the control), but total lipid content slightly decreased. Reducing 75% of basic medium (TAP) N level (0 g/L NaAc + 0.09 g/L NH4Cl treatment) could promote 21.57% total lipid content in comparison with the control (containing 0.38 g/L NH4Cl), but decrease 44.45% biomass and 34.15% lipid productivity. The result of the central composite design (CCD) experiment suggested the optimum total lipid content together with higher biomass and lipid productivity could be obtained under the condition of 4.12 g/L NaAc and 0.20 g/L NH4Cl. They reached 32.14%, 1.68 g/L and 108.21 mg/L/d, and increased by 36.77%, 93.10% and 1.75 times compared with the control, respectively. It suggests moderately increasing C supply and decreasing N levels could synchronously improve the biomass and lipid content of C. reinhardtii.

Low temperature combined with high inoculum density improves alpha-linolenic acid production and biochemical characteristics of Chlamydomonas reinhardtii.

Chlamydomonas reinhardtii grows fast and is rich in polyunsaturated fatty acids. To explore whether the alpha-linolenic acid (ALA) content can be further enhanced, the cultures were incubated under different culture temperatures, light intensities and inoculum densities. Results showed that temperature exhibited more great impact on ALA synthesis of C. reinhardtii than light intensity and inoculum size. The changes of light intensity and inoculum size displayed non-significant effects on ALA content. The optimal ALA proportion in cells was obtained under the condition of 10 °C, 50 µE/m2/s and 5% inoculum density, which reached âˆ¼ 39%.The augmented initial inoculum density could markedly improve the biomass of C. reinhardtii under 10 °C. The maximum ALA productivity (16.42 mg/L/d) was gained under 10 °C coupled with 25% inoculum size, where higher intracellular sugar and protein yield were observed. These results suggest C. reinhardtii would be an alternative feedstock for the industrial production of ALA.

Bioremediation of Pyropia-processing wastewater coupled with lipid production using Chlorella sp.

Pyropia-processing wastewater (PPW) contains diverse organic nutrients and causes environmental pollution. To explore the nutrient removal efficiency and growth performance of Chlorella sp. on PPW, the cultures were conducted in different culture substrates. Results showed that, after 7 days of incubation, the removal rates of total nitrogen (TN), total phosphorus (TP) and phycobiliprotein (PP) all reached more than 90% by cultivating Chlorella sp. C2 and C. sorokiniana F-275 in PPW. The chemical oxygen demand (COD) removal efficiencies could be over 50%. Meanwhile, the increments of biomass in two tested Chlorella strains were 1.39 and 4.89 times higher than those of BG11 and BBM substrates and the increases in lipid productivity were 1.34 and 10.18- fold, respectively. The C18:3 fatty acid proportions were markedly reduced by 27.89% and 29.10%. These results suggest that Chlorella sp. could efficiently reduce various nutrients in PPW and simultaneously accumulate higher biomass with higher biodiesel characteristics.

Correlation among photoluminescence and the electronic and atomic structures of Sr2SiO4:xEu3+ phosphors: X-ray absorption and emission studies.

A series of Eu3+-activated strontium silicate phosphors, Sr2SiO4:xEu3+ (SSO:xEu3+, x = 1.0, 2.0 and 5.0%), were synthesized by a sol-gel method, and their crystalline structures, photoluminescence (PL) behaviors, electronic/atomic structures and bandgap properties were studied. The correlation among these characteristics was further established. X-ray powder diffraction analysis revealed the formation of mixed orthorhombic α'-SSO and monoclinic ß-SSO phases of the SSO:xEu3+ phosphors. When SSO:xEu3+ phosphors are excited under ultraviolet (UV) light (λ = 250 nm, ~ 4.96 eV), they emit yellow (~ 590 nm), orange (~ 613 nm) and red (~ 652 and 703 nm) PL bands. These PL emissions typically correspond to 4f-4f electronic transitions that involve the multiple excited 5D0 → 7FJ levels (J = 1, 2, 3 and 4) of Eu3+ activators in the host matrix. This mechanism of PL in the SSO:xEu3+ phosphors is strongly related to the local electronic/atomic structures of the Eu3+-O2- associations and the bandgap of the host lattice, as verified by Sr K-edge and Eu L3-edge X-ray absorption near-edge structure (XANES)/extended X-ray absorption fine structure, O K-edge XANES and Kα X-ray emission spectroscopy. In the synthesis of SSO:xEu3+ phosphors, interstitial Eu2O3-like structures are observed in the host matrix that act as donors, providing electrons that are nonradiatively transferred from the Eu 5d and/or O 2p-Eu 4f/5d states (mostly the O 2p-Eu 5d states) to the 5D0 levels, facilitating the recombination of electrons that have transitioned from the 5D0 level to the 7FJ level in the bandgap. This mechanism is primarily responsible for the enhancement of PL emissions in the SSO:xEu3+ phosphors. This PL-related behavior indicates that SSO:xEu3+ phosphors are good light-conversion phosphor candidates for use in near-UV chips and can be very effective in UV-based light-emitting diodes.

Bioenergy application of Dunaliella salina SA 134 grown at various salinity levels for lipid production.

The biofuels are receiving considerable attention as a substitute for petro diesel. For microalgae, the cell density or biomass and lipid contents are key components for biodiesel production. This study was conducted to develop favorable culture conditions for Dunaliella salina to maximize its biomass and lipid accumulation. The effect of salinity (0.5 to 2.5 M NaCl) on the cell population, biochemical composition, and lipid output of Dunaliella salina was examined under a controlled environment for 21 days. Maximum growth (6.57 × 107 to 7.17 × 107cells mL-1) potentials were observed at 1.5 to 2 M NaCl. The photosynthetic pigments and carbohydrates also showed trends similar to growth. The maximum carotenoid level (5.16 mg L-1) was recorded at 2 M NaCl. Almost all physicochemical parameters increased with increases in salinity, biomass (1231.66 ± 1.26 mg L-1) and lipid content (248.33 mg L-1), as recorded at 2 M NaCl. Based on fluorescence intensity, the highest values (11.84 × 107cells/ml) of neutral lipids and total lipids (22.28%) were recorded at optimum salinity levels. The present study suggests that a high biomass and lipid accumulation of Dunaliella salina SA 134 could be obtained at the 2 M NaCl level.

Kelp waste extracts combined with acetate enhances the biofuel characteristics of Chlorella sorokiniana.

To probe the effect of kelp waste extracts (KWE) combined with acetate on biochemical composition of Chlorella sorokiniana, the cultures were performed under independent/combined treatment of KWE and acetate. The results showed that high cell density and biomass were obtained by KWE combined with acetate treatments, whose biomass productivity increased by 79.69-102.57% and 20.04-35.32% compared with 3.0gL-1 acetate and KWE treatments respectively. The maximal neutral lipid per cell and lipid productivity were gained in KWE combined with 3.0gL-1 acetate treatment, which increased by 16.32% and 129.03% compared with 3.0gL-1 acetate, and 253.35% and 70.74% compared with KWE treatment. Meanwhile, C18:3n3 and C18:2n6c contents were reduced to 4.90% and 11.88%, whereas C16:0 and C18:1n9c were improved to 28.71% and 37.76%. Hence, supplementing appropriate acetate in KWE cultures is supposed to be a great potential method for large-scale cultivation of C. sorokiniana to generate biofuel.

Effect of Kelp Waste Extracts on the Growth and Development of Pakchoi (Brassica chinensis L.).

To explore the effects of kelp waste extracts (KWE) on the growth and development of Brassia chinensis L., germination and greenhouse experiments were carried out under different concentrations of KWE. The results showed that a higher germination percentage (95%), associated with high germination index (8.70), germination energy (71.67%) and seedling vigor index (734.67), was obtained under a lower KWE concentration (2%) compared with the control. The radicle length (4.97 cm), fresh weight (0.32 g/10 seedlings) and dry weight (0.015 g/10 seedlings) were significantly increased in the treatment of 2% KWE. KWE also could enhance the root growth, the maximum leaf length × width and the fresh weight of plants, the optimal value of which increased by 8.37 cm, 58.14 cm2 and 7.76 g under the treatment of 10% KWE compared with the control respectively. Meanwhile, the contents of vitamin C and soluble sugars in pakchoi leaf were improved by 19.6 mg/100 g and 1.44 mg/g compared with the control, and the nitrate content was decreased by 212.27 mg/kg. Briefly, KWE could markedly stimulate the pakchoi seeds germination at a lower concentration (2%) and enhance the plant growth and quality at a higher concentration (10%).

Effect of kelp waste extracts on the growth and lipid accumulation of microalgae.

Kelp waste extracts (KWE) contained massive soluble sugars, amino acids and various mineral elements. To probe the effects of KWE on microalgal physiological and biochemical responses, the cultures were carried out under the different dilutions. The results showed that 8.0% KWE increased the biomass productivities and total lipid contents of Chlorella strains dramatically, which were 1.83-31.86 times and 20.78-25.91% higher than that of the control. Phaeodactylum tricornutum and Spirulina maxima presented a better growth performance in 1.0% and 4.0% treatment respectively, while their lipid accumulation were not enhanced. In Chlorella-Arc, Chlorella sorokiniana and P. tricornutum, the contents of saturated and monounsaturated fatty acids could be increased, and polyunsaturated fatty acids could be decreased under the conditions of high concentration of KWE (6.0-8.0%). Briefly, KWE facilitated to enhance the biomass productivity and lipid content of Chlorella strains, also improved the fatty acid compositions for biodiesel production.

Breeding of high biomass and lipid producing Desmodesmus sp. by Ethylmethane sulfonate-induced mutation.

To improve the biomass yield and lipid productivity, two desert microalgae, Desmodesmus sp. S81 and G41 were induced mutagenesis by Ethylmethane sulfonate (EMS), and obtained two potential mutants, Desmodesmus sp. S5 and G3 from the mutagenic clones for their greatly promoted biomass and lipid production. The results showed that the biomass yield, lipid content and lipid productivity of the mutant strains S5 and G3 were 778.10mg·L(-1), 48.41% and 19.83mg·L(-1)·d(-1), 739.52mg·L(-1), 46.01%, and 17.92mg·L(-1)·d(-1), respectively, which presented the increment of 45.50%, 8.00% and 74.24%, 20.67%, 10.35% and 55.77% than those of S81 and G41. Comparing with the wild strains, the mutants showed reduced PUFAs and glycol lipids, elevated MUFAs and neutral lipids contents, which were appropriate for biodiesel production.

Growth and lipid accumulation in three Chlorella strains from different regions in response to diurnal temperature fluctuations.

It was economically feasible to screen strains adaptive to wide temperature fluctuation for outdoor cultivation without temperature control. In this research, three Chlorella strains from arctic glacier, desert soil and temperate native lake were isolated and identified. The growth, biochemical composition, lipid content and fatty acid composition of each strain cultured under the mode of diurnal temperature fluctuations were compared. All the three Chlorella strains showed desirable abilities of accumulating lipid under diurnal temperature fluctuations and their fatty acid profiles were suitable for biodiesel production, although the growth and biochemical composition were seemed to be region-specific. The highest lipid content was at 51.83±2.49% DW, 42.80±2.97% DW and 36.13±2.27% DW under different temperature fluctuation of 11 °C, 25 °C, 7 °C, respectively. The results indicated that the three Chlorella strains could be promising biodiesel feedstock for outdoor cultivation by the cultural mode of diurnal temperature fluctuations.