Ultrasonication followed by aqueous two-phase system for extraction, on-site modification and isolation of microalgal starch with reduced digestibility.

Microalgae are new and sustainable sources of starch with higher productivity and flexible production modes than conventional terrestrial crops, but the downstream processes need further development. Here, ultrasonication (with power of 200 W or 300 W and duration of 10, 15, 20, or 25 min) was applied to simultaneously extract and modify starch from a marine microalga Tetraselmis subcordiformis for reducing the digestibility, and an aqueous two-phase system (ATPS) of ethanol/NaH2PO4 was then used to isolate the starches with varied properties. Increasing ultrasonic duration facilitated the partition of starch into the bottom pellet, while enhancing the ultrasonic power was conducive to the allocation in the interphase of the ATPS. The overall starch recovery yield reached 73 âˆ¼ 87 % and showed no significant difference among the ultrasonic conditions tested. The sequential ultrasonication-ATPS process successfully enriched the starch with purities up to 65 % âˆ¼ 88 %, which was among the top levels reported in microalgal starch isolated. Ultrasonication produced more amylose which was mainly fractionated into the interface of the ATPS. The digestibility of the starch was altered under different ultrasonic conditions and varied from different ATPS phases as well, with the one under the ultrasonic power of 200 W for 15 min at the bottom pellet having the highest resistant starch content (RS, 39.7 %). The structural and compositional analysis evidenced that the ultrasonication-ATPS process could exert impacts on the digestibility through altering the surface roughness and fissures of the starch granules, modulating the impurity compositions (protein and lipid) that could interact with starch, and modifying the long- and short-range ordered structures. The developed ultrasonication-ATPS process provided novel insights into the mechanism and strategy for efficient production of functional starch from microalgae with a potential in industrial application.

Nutrient Deprivation Coupled with High Light Exposure for Bioactive Chrysolaminarin Production in the Marine Microalga Isochrysis zhangjiangensis.

Chrysolaminarin, a kind of water-soluble bioactive ß-glucan produced by certain microalgae, is a potential candidate for food/pharmaceutical applications. This study identified a marine microalga Isochrysis zhangjiangensis, in which chrysolaminarin production was investigated via nutrient (nitrogen, phosphorus, or sulfur) deprivations (-N, -P, or -S conditions) along with an increase in light intensity. A characterization of the antioxidant activities of the chrysolaminarin produced under each condition was also conducted. The results showed that nutrient deprivation caused a significant increase in chrysolaminarin accumulation, though this was accompanied by diminished biomass production and photosynthetic activity. -S was the best strategy to induce chrysolaminarin accumulation. An increase in light intensity from 80 (LL) to 150 (HL) µE·m-2·s-1 further enhanced chrysolaminarin production. Compared with -N, -S caused more suitable stress and reduced carbon allocation toward neutral lipid production, which enabled a higher chrysolaminarin accumulation capacity. The highest chrysolaminarin content and concentration reached 41.7% of dry weight (%DW) and 632.2 mg/L, respectively, under HL-S, with a corresponding productivity of 155.1 mg/L/day achieved, which exceeds most of the photoautotrophic microalgae previously reported. The chrysolaminarin produced under HL-N (Iz-N) had a relatively competitive hydroxyl radical scavenging activity at low concentrations, while the chrysolaminarin produced under HL-S (Iz-S) exhibited an overall better activity, comparable to the commercial yeast ß-glucan, demonstrating I. zhangjiangensis as a promising bioactive chrysolaminarin producer from CO2.

Fine-tuned regulation of photosynthetic performance via γ-aminobutyric acid (GABA) supply coupled with high initial cell density culture for economic starch production in microalgae.

Microalgal starch is considered as renewable and sustainable feedstock for biofuels and biorefinery. High cell density culture is favourable for photoautotrophic starch production in microalgae in the aspects of productivity and economy, but it often encounters low starch content or extra stress exposure that limits the production. This study aimed to economically enhance photosynthetic starch production from CO2 fixation in a green microalga Tetraselmis subcordiformis by regulating photosynthetic stress status with a signalling molecule γ-aminobutyric acid (GABA) combined with the application of high initial cell density culture. By increasing initial cell density (ICD) from the normal of 1.1 g L-1 (NICD) to as high as 2.8 g L-1 (HICD), the starch content, yield, and theoretical productivity were improved by 7%, 63%, and 42%, respectively. The addition of GABA under HICD resulted in 14%, 19%, and 26% of further enhancement in starch content, yield, and theoretical productivity, respectively. GABA exhibited distinct regulatory mechanisms on photosynthesis and stress status under HICD relative to NICD. GABA augmented excessive light energy absorption and electron transfer through photosystem II that reinforced the photoinhibition under NICD, while alleviated the stress reversely under HICD, both of which facilitated starch production by enabling a suitable stress status while simultaneously maintaining a sufficient photosynthetic activity. The increase of ICD and/or GABA supply particularly boosted amylopectin accumulation, leading to the changes in starch composition and was more favourable for fermentation-based biofuels production. Preliminary techno-economic analysis showed that the highest net extra benefit of 9.64 $ m-3 culture could be obtained under HICD with 2.5 mM GABA supply where high starch content (62%DW) and yield (2.5 g L-1) were achieved. The combined HICD-GABA regulation was a promising strategy for economic starch production from CO2 by microalgae for sustainable biomanufacturing.

Influence of the use of various imaging units and projections on the radiation dose received by children during chest digital radiography.

OBJECTIVE: To investigate the impact of the use of different imaging units and projections on radiation dose and image quality during chest digital radiography (DR) in 3- and 4-year-old children. METHODS: Two hundred forty 3- and 4-year-old participants requiring chest DR were included; they were divided into three groups: supine anterior-posterior projection (APP), standing APP and standing posterior-anterior projection (PAP). Each group included 40 participants who were evaluated using the same imaging unit. The dose area product (DAP) and the entrance surface dose (ESD) were recorded after each exposure. The visual grading analysis score (VGAS) was used to evaluate image quality, and the longitudinal distance (LD) from the apex of the right lung to the apex of the right diaphragm was used to evaluate the inspiration extent. RESULTS: DAP and ESD were significantly lower in the standing PAP and APP groups than in the supine APP group (P<0.05), but LD was significantly higher in the standing PAP and APP groups than in the supine APP group (P<0.05). Additionally, the pulmonary field area was significantly higher for the standing PAP group than for the standing and supine APP groups (P<0.05). The correlations between ESD, DAP, and VGAS were positive (P<0.001), showing that larger ESD and DAP correspond to higher VGAS. The correlations between ESD, DAP, and body mass index (BMI) were also positive (P<0.05), indicating that higher BMI corresponds to larger ESD and DAP. Finally, no differences in DAP, ESD, VGAS, LD, pulmonary field area, or BMI were noted between males and females (P>0.05). CONCLUSION: The radiation dose to superficial organs may be lower with standing PAP than with standing APP during chest DR. Standing PAP should be selected for chest DR in 3- and 4-year-old children, as it may decrease the required radiation dose.

Effects of Projections on Radiation Dose for and Image Quality of Chest Digital Radiography for Children.

BACKGROUND: Until now few studies have specially validated whether the sex, body mass index, or imaging projections of pediatric patients undergoing chest Digital Radiography (DR) affect the radiation dose and image quality. INTRODUCTION: To investigate the impact of different photography positions on radiation dose for and image quality of chest DR for 3-4-year-old children. METHODS: One-hundred twenty 3-4-year-old patients who required chest DR were included. The patients were divided into 3 groups, with 40 patients in each group: supine Anterior-Posterior Projection (APP), standing APP and posterior-anterior projection (PAP). The Dose Area Product (DAP) and Entrance Surface Dose (ESD) values for every patient were recorded after each exposure. The Visual Grading Analysis Score (VGAS) was used to evaluate image quality. RESULTS: The DAP and ESD values for the standing PAP and APP groups were significantly lower than those for the supine APP group (0.19 ± 0.04 dGy cm2 and 0.05 ± 0.01 mGy vs. 0.25 ± 0.05 dGy cm2 and 0.08 ± 0.01 mGy, P<0.05, respectively). Additionally, the VGAS for the standing APP group was significantly lower than those for the standing PAP and supine APP groups (28.58 ± 0.96 vs. 29.08 ± 0.94 and 29.03 ± 0.80, P<0.05, respectively), whereas the pulmonary field area for the standing PAP group was significantly higher than those for the standing and supine APP groups (118.95 ± 16.81 cm2 vs. 105.65 ± 14.76 cm2 and 105.24 ± 16.32 cm2, P<0.05, respectively). However, there were no statistically significant differences in DAP, ESD, VGAS, pulmonary field area and body mass index between the male and female patients in the three groups (P>0.05, respectively). CONCLUSION: The standing PAP should be the first projection choice for chest DR for 3-4-year-old children; compared with the supine and standing APP, the standing PAP may improve image quality and decrease the required radiation dose.

Technical framework for wastewater-based epidemiology of SARS-CoV-2.

Wastewater-based epidemiology (WBE) is expected to become a powerful tool to monitor the dissemination of SARS-CoV-2 at the community level, which has attracted the attention of scholars all over the world. However, there is not yet a standard protocol to guide its implementation. In this paper, we proposed a comprehensive technical and theoretical framework of relative quantification via qPCR for determining the virus abundance in wastewater and estimating the infection ratio in corresponding communities, which is expected to achieve horizontal and vertical comparability of the data using a human-specific biomarker as the internal reference. Critical factors affecting the virus detectability and the estimation of infection ratio include virus concentration methods, lag-period, per capita virus shedding amount, sewage generation rate, temperature-related decay kinetics of virus/biomarker in wastewater, and hydraulic retention time (HRT), etc. Theoretical simulation shows that the main factors affecting the detectability of virus in sewage are per capita virus shedding amount and sewage generation rate. While the decay of SARS-CoV-2 RNA in sewage is a relatively slow process, which may have limited impact on its detection. Under the ideal condition of high per capita virus shedding amount and low sewage generation rate, it is expected to detect a single infected person within 400,000 people.

Acclimation to a broad range of nitrate strength on a euryhaline marine microalga Tetraselmis subcordiformis for photosynthetic nitrate removal and high-quality biomass production.

Industrial wastewaters usually possess a wide range of nitrate strength. Microalgae-based nitrate-rich wastewater treatment could realize nitrate recovery along with CO2 sequestration for sustainable biomass production, but the low tolerance of the microalgal strains to high-strength nitrate restricted the treatment process. The present study comprehensively evaluated a euryhaline marine microalga Tetraselmis subcordiformis for photosynthetic nitrate removal and biomass production in synthetic wastewater with a broad range of nitrate strength (0.24-7.0 g NO3--N/L). This alga could acclimate to high nitrate strength up to 3.5 g NO3--N/L (HN) without compromising biomass production. Nitrate could be completely removed within four days when low nitrate (0.24 g NO3--N/L, LN) was loaded. The maximum nitrate removal rate of 331 mg N/L/day and specific nitrate removal rate of 360 mg N/day/g cell was obtained under medium nitrate condition (1.8 g NO3--N/L, MN). High-nitrate stress under 7.0 g NO3--N/L (SHN) caused an increased light energy dissipation while decreased the density of photosystem II active reaction center, which partially protect the cells from photodamage and contributed to their acclimation to SHN. The algae also enhanced amino acid/fatty acid proportions essential for maintaining intracellular redox states to cope with the stress caused by LN or SHN. HN and SHN was in favor of protein accumulation and maintenance with enhanced proportion of essential amino acids, which entitled the algal biomass to be of high quality for animal feed applied in livestock graziery and aquaculture. LN facilitated productive starch and lipid accumulation with good quality for biofuels production. The nitrate removal rate and biomass productivity exceeded most of the microalgae reported in literature under similar conditions, which highlighted Tetraselmis subcordiformis as a potent strain for flexible nitrate-rich wastewater remediation coupled with fast CO2 bio-mitigation and high-quality biomass production for sustainable algal biorefinery.

Swine digestate treatment by prior nitrogen-starved Chlorella vulgaris: The effect of over-compensation strategy on microalgal biomass production and nutrient removal.

Anaerobic digester effluent containing high levels of ammonia poses a threat to the environment. To hinder this issue, a modern and promising treatment method incorporates both microalgae and their bioconversion potential. When culturing Chlorella vulgaris at a 1:7 digestate supernatant dilution ratio, biomass concentration was 1.33 g L-1 and 66% of ammonia nitrogen was removed. Furthermore, a prior nitrogen-starved seed method, namely over-compensation strategy, was applied to improve both biomass production and nutrient removal. By using nitrogen-starved seeds after a 48 h nitrogen-free stimulation, biomass yield increased by 1.7-times to 2.56 g L-1. Simultaneously, ammonia nitrogen and total phosphorus removal efficiencies reached 99% and 97% respectively. The enhanced production corresponds to higher chlorophyll fluorescence in the middle and late stages of the culture. In addition, the bioproduct contained 39% carbohydrates, and the proportion of polyunsaturated fatty acids in lipids was 66%. These findings demonstrated that the over-compensation strategy contributed to greater nitrogen removal and high-value bioproduct production in the microalgae-digestate treatment system.

Storage of starch and lipids in microalgae: Biosynthesis and manipulation by nutrients.

Microalgae accumulate starch and lipid as storage metabolites under nutrient depletion, which can be used as sustainable feedstock for biorefinery. Omics analysis coupled with enzymatic and genetic verifications uncovered a partial picture of pathways and important enzymes or regulators related to starch and lipid biosynthesis as well as the carbon partitioning between them under nutrient depletion conditions. Depletion of macronutrients (N, P, and S) resulted in considerable enhancement of starch and/or lipid content in microalgae, but the accompanying declined photosynthesis hampered the achievements of high concentrations. This review summarized the current knowledge on the pathways and the committed steps as well as their carbon allocation involved in starch and lipid biosynthesis, and focused on the manipulation of different nutrients and the alleviation of oxidative stress for enhanced storage metabolites production. The biological and engineering approaches to cope with the conflict between biomass production and storage metabolites accumulation are proposed.

Application of an in situ CO2-bicarbonate system under nitrogen depletion to improve photosynthetic biomass and starch production and regulate amylose accumulation in a marine green microalga Tetraselmis subcordiformis.

BACKGROUND: Microalgal starch is regarded as a promising alternative to crop-based starch for biorefinery such as the production of biofuels and bio-based chemicals. The single or separate use of inorganic carbon source, e.g., CO2 and NaHCO3, caused aberrant pH, which restricts the biomass and starch production. The present study applied an in situ CO2-NaHCO3 system to regulate photosynthetic biomass and starch production along with starch quality in a marine green microalga Tetraselmis subcordiformis under nitrogen-depletion (-N) and nitrogen-limitation (±N) conditions. RESULTS: The CO2 (2%)-NaHCO3 (1 g L-1) system stabilized the pH at 7.7 in the -N cultivation, under which the optimal biomass and starch accumulation were achieved. The biomass and starch productivity under -N were improved by 2.1-fold and 1.7-fold, respectively, with 1 g L-1 NaHCO3 addition compared with the one without NaHCO3 addition. NaHCO3 addition alleviated the high-dCO2 inhibition caused by the single CO2 aeration, and provided sufficient effective carbon source HCO3 - for the maintenance of adequate photosynthetic efficiency and increase in photoprotection to facilitate the biomass and starch production. The amylose content was also increased by 44% under this CO2-bicarbonate system compared to the single use of CO2. The highest starch productivity of 0.73 g L-1 day-1 under -N cultivation and highest starch concentration of 4.14 g L-1 under ±N cultivation were both achieved with the addition of 1 g L-1 NaHCO3. These levels were comparable to or exceeded the current achievements reported in studies. The addition of 5 g L-1 NaHCO3 under ±N cultivation led to a production of high-amylose starch (59.3% of total starch), which could be used as a source of functional food. CONCLUSIONS: The in situ CO2-NaHCO3 system significantly improved the biomass and starch production in T. subcordiformis. It could also regulate the starch quality with varied relative amylose content under different cultivation modes for diverse downstream applications that could promote the economic feasibility of microalgal starch-based biofuel production. Adoption of this system in T. subcordiformis would facilitate the CO2 mitigation couple with its starch-based biorefinery.

Production and structural characterization of a new type of polysaccharide from nitrogen-limited Arthrospira platensis cultivated in outdoor industrial-scale open raceway ponds.

BACKGROUND: Carbohydrates are major biomass source in fuel-targeted biorefinery. Arthrospira platensis is the largest commercialized microalgae with good environmental tolerance and high biomass production. However, the traditional target of A. platensis cultivation is the protein, which is the downstream product of carbohydrates. Aiming to provide the alternative non-food carbohydrates source, the feasible manipulation technology on the cultivation is needed, as well as new separation methodology to achieve maximum utilization of overall biomass. RESULTS: The present study aimed to demonstrate the feasibility of industrially producing carbohydrate-enriched A. platensis and characterize the structure of the polysaccharide involved. Cultivated in industrial-scale outdoor open raceway ponds under nitrogen limitation, A. platensis accumulated maximally 64.3%DW of carbohydrate. The maximum biomass and carbohydrate productivity reached 27.5 g m-2 day-1 and 26.2 g m-2 day-1, respectively. The efficient extraction and purification of the polysaccharides include a high-pressure homogenization-assisted hot water extraction followed by flocculation with a non-toxic flocculant ZTC1 + 1, with the polysaccharide purity and total recovery reaching 81% and 75%, respectively. The purified polysaccharide was mainly composed of (1→3)(1→4)- or (1→3)(1→2)-α-glucan with a molecular weight of 300-700 kDa, which differed from the commonly acknowledged glycogen. CONCLUSIONS: By the way of controlled nitrogen limitation, the high carbohydrate production of A. platensis in the industrial scale was achieved. The α-glucan from A. platensis could be a potential glucose source for industrial applications. A non-toxic separation method of carbohydrate was applied to maintain the possibility of utilization of residue in high-value field.

Phosphorus Enhances Photosynthetic Storage Starch Production in a Green Microalga (Chlorophyta) Tetraselmis subcordiformis in Nitrogen Starvation Conditions.

Microalgae are potential starch producers as alternatives to agricultural crops. This study disclosed the effects and mechanism of phosphorus availability exerted on storage starch production in a starch-producing microalga Tetraselmis subcordiformis in nitrogen starvation conditions. Excessive phosphorus supply facilitated starch production, which differed from the conventional cognition that phosphorus would inhibit transitory starch biosynthesis in plants. Phosphorus enhanced energy utilization efficiency for biomass and storage starch production. ADP-glucose pyrophosphorylase (AGPase), conventionally known to be critical for starch biosynthesis, was negatively correlated to storage starch biosynthesis. Excessive phosphorus supply maintained large cell volumes, enhanced activities of starch phosphorylases (SPs) along with branching enzymes and isoamylases, and increased phosphoenolpyruvate and trehalose-6-phosphate levels to alleviate the inhibition of high phosphate availability to AGPase, all of which improved starch production. This work highlighted the importance of phosphorus in the production of microalgal starch and provided further evidence for the SP-based storage starch biosynthesis pathway.

The ΔF/Fm'-guided supply of nitrogen in culture medium facilitates sustainable production of TAG in Nannochloropsis oceanica IMET1.

BACKGROUND: Triacylglycerol (TAG) from photosynthetic microalgae is a sustainable feedstock for biodiesel production. Physiological stress triggers microalgal TAG accumulation. However excessive physiological stress will impair the photosynthesis system seriously thus decreasing TAG productivity because of the low biomass production. Hence, it is critical to quantitatively and timely monitor the degree of the stress while the microalgal cells growing so that the optimal TAG productivity can be obtained. RESULTS: The lack of an on-line monitored indicator has limited our ability to gain knowledge of cellular "health status" information regarding high TAG productivity. Therefore, to monitor the degree of nitrogen stress of the cells, we investigated the correlation between the photosynthetic system II (PS II) quantum yield and the degree of stress based on the high relevancy between photosynthetic reduction and nitrogen stress-induced TAG accumulation in microalgal cells. ΔF/Fm', which is the chlorophyll fluorescence parameter that reflects the effective capability of PS II, was identified to be a critical factor to indicate the degree of stress of the cells. In addition, the concept of a nitrogen stress index has been defined to quantify the degree of stress. Based on this index and by monitoring ΔF/Fm' and guiding the supply of nitrogen in culture medium to maintain a stable degree of stress, a stable and efficient semi-continuous process for TAG production has been established. CONCLUSION: The results indicate that the semi-continuous cultivation process with a controlled degree of stress by monitoring the ΔF/Fm' indicator will have a significant impact on microalgal TAG production, especially for the outdoor controllable cultivation of microalgae on a large scale.

Characterization of starch phosphorylase from the marine green microalga (Chlorophyta) Tetraselmis subcordiformis reveals its potential role in starch biosynthesis.

In a marine green starch-producing microalga Tetraselmis subcordiformis, the role of starch phosphorylase (SP) in the starch biosynthesis was disclosed by characterizing the enzyme properties and activity variations during the starch accumulation process. TsSP4, a SP isoform accounting for the major SP activity in T. subcordiformis, was unique to be active in a monomer form with a molecular weight of approximately 110kDa. It resembled one of the chloroplast-located SPs (PhoA) in Chlamydomonas reinhardtii with a similarity of 63.3% in sequence, though it possessed the typical L78/80 domain found in the plastidial SPs (Pho1) of higher plants that was absent in PhoA. TsSP4 exhibited moderate sensitivity to ADP-Glc inhibition and had a high activity for longer-chain linear maltooligosacchride (MOS) and amylopectin against highly branched glycogen as the substrates. TsSP4 had 2-fold higher affinity for Glc-1-P in the synthetic direction than for Pi in the phosphorolytic direction, and the catalytic constant kcat for Glc-1-P was 2-fold of that for Pi. Collectively, TsSP4 preferred synthetic rather than phosphorolytic direction. TsSP4 could elongate MOSs even initially with Pi alone in the absence of Glc-1-P, which further supported its synthetic role in the starch biosynthesis. TsSP4 displayed increased activities in the developing and mature stage of starch biosynthesis under nitrogen-starvation conditions, indicating its possible contribution to the amylopectin amplification.

FabG can function as PhaB for poly-3-hydroxybutyrate biosynthesis in photosynthetic cyanobacteria Synechocystis sp. PCC 6803.

The production of poly-3-hydroxybutyrate (PHB) by photosynthetic cyanobacteria is a potentially sustainable production method for the biodegradable plastics industry. ß-Ketoacyl-ACP reductase (FabG), from the photosynthetic cyanobacterium Synechocystis sp. PCC 6803 (SpFabG), is the first NADPH-dependent reductase in the fatty acid biosynthesis pathway. Its structure is similar to that of acetoacetyl-CoA reductase (SpPhaB), which is critical for PHB synthesis and can replace SpPhaB for acetoacetyl-CoA reduction in vitro. However, the specific function of SpFabG in fatty acid synthesis and whether SpFabG could participate in PHB synthesis in vivo were not yet clear. In this study, the role of SpFabG in fatty acid synthesis was first verified in vivo by knocking down and overexpressing of fabG. It was shown that SpFabG was essential yet not rate-limiting for fatty acid biosynthesis. The biochemical characterization of SpFabG using acetoacetyl-CoA as the substrate showed that the optimum temperature, optimum pH, Km and kcat were 30°C, 7, 2.30 mM, and 19.85 s-1, respectively, which exemplified the ability of SpFabG to reduce acetoacetyl-CoA with a relatively low affinity and weak catalytic efficiency. Functional analysis of SpFabG in vivo indicated that SpFabG was able to partially complement SpPhaB under nitrogen-deprived conditions, and overexpression of fabG led to the diversion of partial carbon flux from fatty acid toward PHB synthesis.

Evaluation of the integrated hydrothermal carbonization-algal cultivation process for enhanced nitrogen utilization in Arthrospira platensis production.

Sustainable microalgal cultivation at commercial scale requires nitrogen recycling. This study applied hydrothermal carbonization to recover N of hot-water extracted Arthrospira platensis biomass residue into aqueous phase (AP) under different operation conditions and evaluated the N utilization, biomass yield and quality of A. platensis cultures using AP as the sole N source. With the increase of temperature at 190-210°C or reaction time of 2-3h, the N recovery rate decreased under nitrogen-repletion (+N) cultivation, while contrarily increased under nitrogen-limitation (-N) cultivation. Under +N biomass accumulation in the cultures with AP under 190°C was enhanced by 41-67% compared with that in NaNO3, and the highest protein content of 51.5%DW achieved under 200°C-2h was also 22% higher. Carbohydrate content of 71.4%DW under -N cultivation achieved under 210°C-3h was 14% higher than that in NaNO3. HTC-algal cultivation strategy under -N mode could save 60% of conventional N.

Utilization of recovered nitrogen from hydrothermal carbonization process by Arthrospira platensis.

In the context of sustainable cultivation of microalgae, the present study focused on the use of nitrogen from the hot-water extracted biomass residue of Arthrospira platensis by hydrothermal carbonization (HTC) and the sequential cultivation of the same alga with the HTC aqueous phase (AP). Nearly 90% of the nitrogen recovered from HTC into AP was in the organic form. Under nitrogen-limited condition with HTCAP as nitrogen source the yield and content of carbohydrate were enhanced by 21% and 15% respectively compared with that under the same nitrogen level provided by NaNO3, which entitled HTCAP for the substitution of conventional nitrate. In the same way pilot-scale cultivation of A. platensis in raceway ponds outdoors demonstrated that carbohydrate content of 43.8% DW and productivity of 10.3g/m(2)/d was achieved. Notably 54% of organic nitrogen in the HTCAP could be recycled by cultivation of pre-nitrogen starved A. platensis as seeds under nitrogen limitation.

Coordinated regulation of nitrogen supply mode and initial cell density for energy storage compounds production with economized nitrogen utilization in a marine microalga Isochrysis zhangjiangensis.

Lipids and carbohydrates are main energy storage compounds (ESC) of microalgae under stressed conditions and they are potential feedstock for biofuel production. Yet, the sustainable and commercially successful production of ESC in microalgae needs to consider nitrogen utilization efficiency. Here the impact of different initial cell densities (ICDs) on ESC accumulation in Isochrysis zhangjiangensis under two nitrogen supply modes (an initially equal concentration of nitrogen per-cell in the medium (N1) and an equal total concentration of nitrogen in the culture system (N2)) were investigated. The results demonstrated that the highest ESC yield (1.36gL(-1)) at N1, which included a maximal nitrogen supply in the cultivation system, and the highest ESC content (66.5%) and ESC productivity per mass of nitrogen (3.28gg(-1) (N) day(-1)) at N2, were all obtained under a high ICD of 8.0×10(6)cellsmL(-1). Therefore I. zhangjiangensis qualifies for ESC-enriched biomass production with economized nitrogen utilization.

Investigating Cellular Responses During Photohydrogen Production by the Marine Microalga Tetraselmis subcordiformis by Quantitative Proteome Analysis.

The marine microalga Tetraselmis subcordiformis could photoproduce hydrogen under the regulation of carbonyl cyanide m-chlorophenylhydrazone (CCCP), and a hydrogen production process kinetic analysis was characterized by two peaks, suggesting that two distinct mechanisms might exist in this alga. Therefore, 2D nanoliquid chromatography-tandem mass spectrometry (LC-MS/MS) was introduced to analyze the proteome of samples from different time points. A total of 912 proteins were identified, providing a global view of the cellular responses at the proteomic level. These proteins can be divided into multiple functional groups including stress responses, energy metabolism and redox homeostasis. The quantitative proteomic data provided more details on the electron donors for hydrogen production. During the first stage, photosystem II produced electrons for hydrogen production; during the second stage, metabolites were the major electron donors via nonphotochemical plastoquinone reduction by NADH dehydrogenase.

Identification of carbohydrates as the major carbon sink of the marine microalga Isochrysis zhangjiangensis (Haptophyta) and optimization of its productivity by nitrogen manipulation.

Microalgae represent a potential feedstock for biofuel production. During cultivation under nitrogen-depleted conditions, carbohydrates, rather than neutral lipids, were the major carbon sink of the marine microalga Isochrysis zhangjiangensis (Haptophyta). Carbohydrates reached maximum levels of 21.2 pg cell(-1) on day 5, which was an increase of more than 7-fold from day 1, while neutral lipids simultaneously increased 1.9-fold from 4.0 to 7.6 pg cell(-1) during the ten-day nitrogen-depleted cultivation. The carbohydrate productivity of I. zhangjiangensis was improved by optimization of the nitrate supply mode. The maximum carbohydrate concentration was 0.95 g L(-1) under batch cultivation, with an initial nitrogen concentration of 31.0 mg L(-1), which was 2.4-fold greater than that achieved under nitrogen-depleted conditions. High performance liquid chromatography (HPLC) analysis showed that the accumulated carbohydrate in I. zhangjiangensis was composed of glucose. These results show that I. zhangjiangensis represents an ideal carbohydrate-enriched bioresource for biofuel production.