Photosynthesis and biochemical characterization of the green alga Chlamydopodium fusiforme (Chlorophyta) grown in a thin-layer cascade.

Photosynthesis, growth and biochemical composition of the biomass of the freshwater microalga Chlamydopodium fusiforme cultures outdoors in a thin-layer cascade were investigated. Gross oxygen production measured off-line in samples taken from the outdoor cultures was correlated with the electron transport rate estimated from chlorophyll a fluorescence measurements. According to photosynthesis measurements, a mean of 38.9 ± 10.3 mol of photons were required to release one mole of O2, which is 4.86 times higher than the theoretical value (8 photons per 1 O2). In contrast, according to the fluorescence measurements, a mean of 11.7 ± 0.74 mol of photons were required to release 1 mol of O2. These findings indicate that fluorescence-based photosynthesis rates may not be fully replace oxygen measurements to evaluate the performance of an outdoor culture. Daily gross biomass productivity was 0.3 g DW L-1 day-1 consistently for 4 days. Biomass productivity was strongly affected by the suboptimal concentration at which the culture was operated and by the respiration rate, as the substantial volume of culture was kept in the dark (about 45% of the total volume). As the cells were exposed to excessive light, the photosynthetic activity was mainly directed to the synthesis of carbohydrates in the biomass. In the morning, carbohydrate content decreased because of the dark respiration. Per contra, protein content in the biomass was lower at the end of the day and higher in the morning due to carbohydrate consumption by respiration. The data gathered in these trials are important for the future exploitation of Chlamydopodium fusiforme as a potential novel species in the field of microalgae for the production of bio-based compounds.

Growth and photosynthetic performance of Nostoc linckia (formerly N. calcicola) cells grown in BG11 and BG110 media.

The biotechnological potential of Nostoc linckia as a biofertilizer and source of bioactive compounds makes it important to study its growth physiology and productivity. Since nitrogen is a fundamental component of N. linckia biomass, we compared the growth and biochemical composition of cultures grown in BG11 (i.e., in the presence of nitrate) and BG110 (in the absence of nitrate). Cultures grown in BG11 accumulated more cell biomass reaching a dry weight of 1.65 ± 0.06 g L-1, compared to 0.92 ± 0.01 g L-1 in BG110 after 240 h of culture. Biomass productivity was higher in culture grown in BG11 medium (average 317 ± 38 mg L-1 day-1) compared to that attained in BG110 (average 262 ± 37 mg L-1 day-1). The chlorophyll content of cells grown in BG11 increased continuously up to (39.0 ± 1.3 mg L-1), while in BG110 it increased much more slowly (13.6 ± 0.8 mg L-1). Biomass grown in BG11 had higher protein and phycobilin contents. However, despite the differences in biochemical composition and pigment concentration, between BG11 and BG110 cultures, both their net photosynthetic rates and maximum quantum yields of the photosystem II resulted in similar.

Investigation of Photosystem II Functional Size in Higher Plants under Physiological and Stress Conditions Using Radiation Target Analysis and Sucrose Gradient Ultracentrifugation.

The photosystem II (PSII) reaction centre is the critical supramolecular pigment-protein complex in the chloroplast which catalyses the light-induced transfer of electrons from water to plastoquinone. Structural studies have demonstrated the existence of an oligomeric PSII. We carried out radiation inactivation target analysis (RTA), together with sucrose gradient ultracentrifugation (SGU) of PSII, to study the functional size of PSII in diverse plant species under physiological and stress conditions. Two PSII populations, made of dimeric and monomeric core particles, were revealed in Pisum sativum, Spinacea oleracea, Phaseulus vulgaris, Medicago sativa, Zea mais and Triticum durum. However, this core pattern was not ubiquitous in the higher plants since we found one monomeric core population in Vicia faba and a dimeric core in the Triticum durum yellow-green strain, respectively. The PSII functional sizes measured in the plant seedlings in vivo, as a decay of the maximum quantum yield of PSII for primary photochemistry, were in the range of 75-101 ± 18 kDa, 2 to 3 times lower than those determined in vitro. Two abiotic stresses, heat and drought, imposed individually on Pisum sativum, increased the content of the dimeric core in SGU and the minimum functional size determined by RTA in vivo. These data suggest that PSII can also function as a monomer in vivo, while under heat and drought stress conditions, the dimeric PSII structure is predominant.

Purple non-sulfur bacteria as cell factories to produce a copolymer as PHBV under light/dark cycle in a 4-L photobioreactor.

The present study reports a strategy to produce polyhydroxyalkanoates (PHAs) by culturing the marine bacterium Rhodovulum sulfidophilum DSM-1374. The study was carried out by growing the bacterium anaerobically for 720 h under 16/8 light/dark cycle. Two analytical techniques such as proton magnetic nuclear magnetic resonance (1H NMR) and Fourier transform infrared spectroscopy (FT-IR) were used to determine that the polyester produced was poly-3-hydroxybutirate-co-3-hydroxyvalerate (PHBV). This study showed that the excess of lactate and the limitation of N-P nutrients under a light-dark cycle enhanced PHBV synthesis and achieved a PHBV concentration of 330 mg/L in the R. sulfidophilum culture. During the 30 days of bacterial cultivation, the percentage of polymer in the six harvested dry biomasses gradually increased from 13.7% to 23.4%. In addition, the study showed that PHBV synthesis stopped during the 8-h dark phase and restarted in the light. The light-dark cycle study also showed that R. sulfidophilum DSM-1374 can be grown outdoors because the cells are exposed to the natural light-dark cycle.

Effect of plate distance on light conversion efficiency of a Synechocystis culture grown outdoors in a multiplate photobioreactor.

In this work, the performance of a vertical multiplate photobioreactor is analyzed and presented. The photobioreactor consisted of 20 vertical plates (1 m2 each) connected by manifolds and a working volume of 1300 L. The total area occupied (footprint) was 10 m2, while the illuminated area was 40 m2, therefore the ratio of illuminated area to volume ratio was about 30 m-1. The performance of the photobioreactor was evaluated using a culture of Synechocystis PCC 6803, circulated by a centrifuge pump. The results showed that the amount of light captured by the photobioreactor at a plate spacing of 0.5 m was 90.2 % of the light incident on the horizontal surface, while at a plate spacing of 1.0 m, 50.3 % was captured. The corresponding biomass yield, calculated based on the ground area occupied by the reactor, was 26.0 g m-2 day-1 and 7.2 g m-2 day-1, when the plates were spaced at 0.5 m and 1.0 m respectively. Therefore, the light conversion efficiency calculated based on the ground area was significantly higher in the configuration with a plate spacing of 0.5 m, reaching 5.43 % based on PAR (photosynthetically active radiation), and 2.44 % based on solar radiation, giving a value 3.7 higher than when the plates were spaced 1.0 m apart. It was concluded that the light conversion efficiency might be further improved by reducing the plate spacing while also reducing the culture light path.

In situ monitoring of chlorophyll a fluorescence in Nannochloropsis oceanica cultures to assess photochemical changes and the onset of lipid accumulation during nitrogen deprivation.

In situ chlorophyll a fluorescence measurements were applied to monitor changes in the photochemical variables of Nannochloropsis oceanica cultures under nitrogen-deplete and nitrogen-replete (control) conditions. In addition, growth, lipid, fatty acid, and pigment contents were also followed. In the control culture, growth was promoted along with pigment content, electron transport rate (ETR), and polyunsaturated fatty acids, while total lipid content and fatty acid saturation level diminished. Under nitrogen-deplete conditions, the culture showed a higher de-epoxidation state of the xanthophyll cycle pigments. Fast transients revealed a poor processing efficiency for electron transfer beyond QA , which was in line with the low ETR due to nitrogen depletion. Lipid content and the de-epoxidation state were the first biochemical variables triggered by the change in nutrient status, which coincided with a 20% drop in the in situ effective quantum yield of PSII (ΔF'/Fm '), and a raise in the Vj measurements. A good correlation was found between the changes in ΔF'/Fm ' and lipid content (r = -0.96, p < 0.01). The results confirm the reliability and applicability of in situ fluorescence measurements to monitor lipid induction in N. oceanica.

Photoautotrophs-Bacteria Co-Cultures: Advances, Challenges and Applications.

Photosynthetic microorganisms are among the fundamental living organisms exploited for millennia in many industrial applications, including the food chain, thanks to their adaptable behavior and intrinsic proprieties. The great multipotency of these photoautotroph microorganisms has been described through their attitude to become biofarm for the production of value-added compounds to develop functional foods and personalized drugs. Furthermore, such biological systems demonstrated their potential for green energy production (e.g., biofuel and green nanomaterials). In particular, the exploitation of photoautotrophs represents a concrete biorefinery system toward sustainability, currently a highly sought-after concept at the industrial level and for the environmental protection. However, technical and economic issues have been highlighted in the literature, and in particular, challenges and limitations have been identified. In this context, a new perspective has been recently considered to offer solutions and advances for the biomanufacturing of photosynthetic materials: the co-culture of photoautotrophs and bacteria. The rational of this review is to describe the recently released information regarding this microbial consortium, analyzing the critical issues, the strengths and the next challenges to be faced for the intentions attainment.

Production of Biohydrogen and/or Poly-ß-hydroxybutyrate by Rhodopseudomonas sp. Using Various Carbon Sources as Substrate.

The polyhydroxyalkanoates (PHA) are family of biopolyesters synthesized by numerous bacteria which are attracting a great attention due to their thermoplastic properties. Polyhydroxybutyrate (PHB) is the most common type of PHA which presents thermoplastic and biodegradable properties. It is synthesized under stressful conditions by heterotrophic bacteria and many photosynthetic microorganisms such as purple non-sulfur bacteria and cyanobacteria. Biological hydrogen (H2) production is being evaluated for use as a fuel since it is a promising substitute for carbonaceous fuels owing to its high conversion efficiency and high specific content. In the present work, the purple non-sulfur photosynthetic bacterium Rhodopseudomonas sp. for the simultaneous H2 photo-evolution and poly-ß-hydroxybutyrate (PHB) production has been investigated. Three different types of carbon sources were tested in the presence of glutamate as a nitrogen source in a batch cultivation system, under continuous irradiance. The results indicated the fact that the type of carbon source in the culture broth affects in various ways the metabolic activity of the bacterial biomass, as evidenced by the production of PHB and/or H2 and biomass. The best carbon source for PHB accumulation and H2 production by Rhodopseudomonas sp. turned out to be the acetate, having the highest H2 production (2286 mL/L) and PHB accumulation (68.99 mg/L, 18.28% of cell dry weight).

Bioplastic production by feeding the marine Rhodovulum sulfidophilum DSM-1374 with four different carbon sources under batch, fed-batch and semi-continuous growth regimes.

In the present study, the ability of the marine bacterium Rhodovulum sulfidophilum DSM-1374 to convert, via photo-fermentative process, certain organic acids such as single carbon source (acetate, lactate, malate and succinate) into polyhydroxyalkanoate accumulations within bacterial cells is evaluated. The main goal of the investigation was poly-3-hydroxybutyrate (P3HB) synthesis by a photo-fermentative process. Of the four carbon sources, only succinate simultaneously produced P3HB and H2 (268 mg/L and 1085 mL/L respectively). Malate was the least productive source for P3HB; the other carbon sources (acetate and lactate) produced a significant amount of polymer (596 mg P3HB/L for acetate and 716 mg P3HB/L for lactate) when R. sulfidophilum was cultured in batch growth conditions. Cumulative P3HB increased significantly when the bacterium was grown under two steps: nutrient sufficient conditions (step 1) followed by macronutrient deficient conditions (step 2). The highest cumulative P3HB was observed at the end of step 2 (1000 mg/L) when R. sulfidophilum was fed with lactate under phosphorus starvation. When grown over 1200 h, under a semi-continuous regimen, the harvested dry-biomass reached a constant content of P3HB (39.1 ± 1.6 % of cell dry-weight), in the semi-steady state condition. Since lactate is an abundant byproduct of world industries, it can be used to mitigate the environmental impact in a modern circular bio-economy.

Effects of pH, temperature and salinity on P3HB synthesis culturing the marine Rhodovulum sulfidophilum DSM-1374.

Rhodovulum sulfidophilum DSM-1374 is a potential producer of polyester when growing in phototrophic conditions. The present study investigated on a polyester product (P3HB) by culturing Rhodovulum sulfidophilum DSM-1374 in two different photobioreactors (PBR-1 and PBR-2) both with 4-L working volumes. PBR-1 is equipped with an internal rotor having 4 paddles to mix the bacterial culture while PBR-2 has an internal coil-shaped rotor. After selecting PBR-1, which best performed in the preliminary experiment, the effect of different stressing growth conditions as pH (7.0, 8.0, and 9.0), temperature (25, 30, and 35 °C), and medium salinity (1.5, 2.5, 3.5, and 4.5%) were tested. When the pH of the culture was set to 8.0, the capability of the bacterium to synthetize the polyester increased significantly reaching a concentration of 412 mg (P3HB)/L; the increase of the pH at 9.0 caused a reduction of the P3HB concentration in the culture. The medium salinity of 4.5% was the best stress-growth condition to reach the highest concentration of polyester in the culture (820 ± 50 mg (P3HB)/L) with a P3HB mass fraction in the dry biomass of 33 ± 1.5%. Stresses caused by culture temperature are another potential parameter that could increase the synthesis of P3HB.

Whey and molasses as inexpensive raw materials for parallel production of biohydrogen and polyesters via a two-stage bioprocess: New routes towards a circular bioeconomy.

Consecutive dark-fermentation and photo-fermentation stages were investigated for a profitable circular bio-economy. H2 photo-production versus poly(3-hydroxybutyrate) (P3HB) accumulation is a modern biotechnological approach to use agro-food industrial byproducts as no-cost rich-nutrient medium in eco-sustainable biological processes. Whey and molasses are very important byproducts rich in nutrients that lactic acid bacteria can convert, by dark-fermentation, in dark fermented effluents of whey (DFEW) and molasses (DFEM). These effluents are proper media for culturing purple non-sulfur bacteria, which are profitable producers of P3HB and H2. The results of the present study show that Lactobacillus sp. and Rhodopseudomonas sp. S16-VOGS3 are two representative genera for mitigation of environmental impact. The highest productivity of P3HB (4.445 mg/(L·h)) was achieved culturing Rhodopseudomonas sp. S16-VOGS3, when feeding the bacterium with 20% of DFEM; the highest H2 production rate of 4.46 mL/(L·h) was achieved when feeding the bacterium with 30% of DFEM.

Poly-3-hydroxybutyrate and H2 production by Rhodopseudomonas sp. S16-VOGS3 grown in a new generation photobioreactor under single or combined nutrient deficiency.

The main goal of this investigation was setting up a growth strategy to separate H2 evolution from P3HB synthesis in order to increase cumulative P3HB in Rhodopseudomonas cells. The accumulation of poly-3-hydroxybutyrate (P3HB) was investigated culturing Rhodopseudomonas sp. S16-VOGS3 with three carbon substrates either as acetate, butyrate or lactate and with two nitrogen sources either as ammonium or glutamate. The investigation was carried out under several stress conditions caused by single or double nutrient deficiency. The content of P3HB in cell dry weight (CDW) was 21.8% with lactate; 24.6% with acetate and 27.6% with butyrate under sulfur deficient conditions. The P3HB content increased significantly culturing Rhodopseudomonas sp. S16-VOGS3 with butyrate following three phases of growth: phase-1, nutrient sufficient conditions; phase-2, nitrogen-deficiency and phase-3, sulfur-deficient conditions. Under this last phase, the highest P3HB content was achieved (34.4% of CDW). A combined production of P3HB and molecular H2 was obtained when Rhodopseudomonas sp. S16-VOGS3 was cultured with either acetate or butyrate under nitrogen sufficiency (glutamate) or nitrogen deficiency.

Effect of high pH on growth of Synechocystis sp. PCC 6803 cultures and their contamination by golden algae (Poterioochromonas sp.).

Culturing cyanobacteria in a highly alkaline environment is a possible strategy for controlling contamination by other organisms. Synechocystis PCC 6803 cells were grown in continuous cultures to assess their growth performance at different pH values. Light conversion efficiency linearly decreased with the increase in pH and ranged between 12.5 % (PAR) at pH 7.5 (optimal) and decreased to 8.9 % at pH 11.0. Photosynthetic activity, assessed by measuring both chlorophyll fluorescence and photosynthesis rate, was not much affected going from pH 7.5 to 11.0, while productivity, growth yield, and biomass yield on light energy declined by 32, 28, and 26 % respectively at pH 11.0. Biochemical composition of the biomass did not change much within pH 7 and 10, while when grown at pH 11.0, carbohydrate content increased by 33 % while lipid content decreased by about the same amount. Protein content remained almost constant (average 65.8 % of dry weight). Cultures maintained at pH above 11.0 could grow free of contaminants (protozoa and other competing microalgae belonging to the species of Poterioochromonas).

A bioenergetic assessment of photosynthetic growth of Synechocystis sp. PCC 6803 in continuous cultures.

BACKGROUND: Synechocystis sp. PCC 6803, a model organism used for bioenergy and bioplastic production, was grown in continuous culture to assess its most important bioenergetic parameters. RESULTS: Biomass yield on light energy of 1.237 g mol photons(-1) and maintenance energy requirement of 0.00312 mol photons g(-1) h(-1) were calculated. This corresponded to a light conversion efficiency of 12.5 %, based on the model of Pirt which was about 35 % lower than the theoretical one based on the stoichiometric equation for the formation of biomass on carbon dioxide, water, and nitrate. The maximum F v/F m ratio recorded in the Synechocystis cultures was 0.57; it progressively declined to 0.45 as the dilution rate increased. An over-reduction of reaction centers at a high dilution rate was also recorded, together with an increased VJ phase for the chlorophyll fluorescence transient. In contrast, the chlorophyll optical cross section increased by about 40 % at the fastest dilution rate, and compensated for the decline in F v/F m, thus resulting in a constant total photosynthesis rate (photosynthesis plus respiration). Chlorophyll content was maximum at the lowest dilution rate and was 48 % lower at the highest one, while phycocyanin, and total carotenoids decreased by about 42 % and 37 %, respectively. Carotenoid analysis revealed increased echinenone, zeaxanthin, and myxoxanthophyll contents as the dilution rate increased (40.6, 63.8 and 35.5 %, respectively, at the fastest dilution rate). A biochemical analysis of the biomass harvested at each different dilution rates showed no changes in the lipid content (averaging 11.2 ± 0.6 % of the dry weight), while the protein content decreased as the dilution rate increased, ranging between 60.7 ± 1.1 and 72.6 ± 0.6 %. Amino acids pattern did not vary with the dilution rate. Carbohydrate content ranged from 9.4 to 16.2 % with a mean value of 11.2 ± 1.4 %. CONCLUSIONS: The present work provides useful information on the threshold of light conversion efficiency in Synechocystis, as well as basic bioenergetic parameters that will be helpful for future studies related to its genetic transformation and metabolic network reconstruction.

Preventive or potential therapeutic value of nutraceuticals against ionizing radiation-induced oxidative stress in exposed subjects and frequent fliers.

Humans are constantly exposed to ionizing radiation deriving from outer space sources or activities related to medical care. Absorption of ionizing radiation doses over a prolonged period of time can result in oxidative damage and cellular dysfunction inducing several diseases, especially in ageing subjects. In this report, we analyze the effects of ionizing radiation, particularly at low doses, in relation to a variety of human pathologies, including cancer, and cardiovascular and retinal diseases. We discuss scientific data in support of protection strategies by safe antioxidant formulations that can provide preventive or potential therapeutic value in response to long-term diseases that may develop following exposure.

A photosynthetic biosensor with enhanced electron transfer generation realized by laser printing technology.

One of the limits of current electrochemical biosensors is a lack of methods providing stable and highly efficient junctions between biomaterial and solid-state devices. This paper shows how laser-induced forward transfer (LIFT) can enable efficient electron transfer from photosynthetic biomaterial immobilized on screen-printed electrodes (SPE). The ideal pattern, in terms of photocurrent signal of thylakoid droplets giving a stable response signal with a current intensity of approximately 335 ± 13 nA for a thylakoid mass of 28 ± 4 ng, was selected. It is shown that the efficiency of energy production of a photosynthetic system can be strongly enhanced by the LIFT process, as demonstrated by use of the technique to construct an efficient and sensitive photosynthesis-based biosensor for detecting herbicides at nanomolar concentrations.

Intercalation of the herbicide atrazine in layered double hydroxides for controlled-release applications.

BACKGROUND: The herbicide atrazine was intercalated in the interlayer region of Mg/Al layered double hydroxides in order to produce a nanohybrid that could be used in controlled-release applications. RESULTS: The hydrophobic herbicide was incorporated in artificial membranes formed in the interlayer of the inorganic host by using palmitic acid. The synthetic nanohybrid material was characterised by various techniques, and release studies were carried out. In addition, the photosynthetic alga Chlamydomonas reinhardtii Dang. was treated with the atrazine-containing nanohybrid, which exerted an herbicidal efficacy similar to that of the free herbicide. CONCLUSION: The herbicide containing nanohybrid could enable a controlled release of the herbicide. In addition, the herbicide would be delivered close to its site of uptake, enhancing efficiency and reducing the required doses.

Nutraceutical use of garlic sulfur-containing compounds.

Garlic is one of the world's oldest medicines that has been employed not only for flavouring but also as a medical herb for its prophylactic and therapeutic actions. Most garlics' beneficial effects are due to the presence of the organosulphate molecule allicin. Allicin is a highly unstable molecule and, during processing, is rapidly transformed into a variety of organosulfur components. The enzyme alliinase, which is responsible for the conversion of alliin to allicin, is irreversibly destroyed at the acidic environment of stomach. This is the reason why most garlic supplements contain garlic powder or granules, but do not contain allicin itself. Garlic alliinase could be encapsulated and coated with materials which would protect it in the harsh conditions of the stomach. The objective of this chapter is to summarize the most important garlic health benefits and to discuss promising encapsulation/stabilization approaches.

Development of a photosystem II-based optical microfluidic sensor for herbicide detection.

Herbicides are highly toxic for both human and animal health. The increased application of herbicides in agriculture during the last decades has resulted in the contamination of both soil and water. Herbicides, under illumination, can inhibit photosystem II electron transfer. Photosynthetic membranes isolated from higher plants and photosynthetic micro-organisms, immobilized and stabilized, can serve as a biorecognition element for a biosensor. The inhibition of photosystem II causes a reduced photoinduced production of hydrogen peroxide, which can be measured by a chemiluminescence reaction with luminol and the enzyme horseradish peroxidase. In the present work, a compact and portable sensing device that combines the production and detection of hydrogen peroxide in a single flow assay is proposed for herbicide detection.