The growth of <Emphasis Type="Italic">Chlorella sorokiniana</Emphasis> as influenced by CO<Subscript>2</Subscript>, light,and flue gases

In order to achieve recognition as environmentally friendly production, flue gases should be used as a CO₂ source for growing the microalgae Chlorella sorokiniana when used for hydrogen production. Flue gases from a waste incinerator and from a silicomanganese smelter were used. Before testing the flue gases, the algae were grown in a laboratory at 0.04, 1.3, 5.9, and 11.0 % (v/v) pure CO₂ gas mixed with fresh air. After 5 days of growth, the dry biomass per liter algal culture reached its maximum at 6.1 % CO₂. A second experiment was conducted in the laboratory at 6.2 % CO₂ at photon flux densities (PFD) of 100, 230, and 320 μmol photons m^?2 s^?1. After 4 days of growth, increasing the PFD increased the biomass production by 67 and 108 % at the two highest PFD levels, as compared with the lowest PFD. A bioreactor system containing nine daylight-exposed tubes and nine artificial light-exposed tubes was installed on the roof of the waste incinerator. The effect of undiluted flue gas (10.7 % CO₂, 35.8 ppm NO _x , and 38.6 ppm SO₂), flue gas diluted with fresh air to give 4.2 % CO₂ concentration, and 5.0 % pure CO₂ gas was studied in daylight (21.4?±?9.6 mol photons m^?2 day^?1 PAR, day length 12.0 h) and at 135 μmol photons m^?2 s^?1 artificial light given 24 h day^?1 (11.7?±?0.0 mol photons m^?2 day^?1 PAR). After 4 days’ growth, the biomass production was the same in the two flue gas concentrations and the 5 % pure CO₂ gas control. The biomass production was also the same in daylight and artificial light, which meant that, in artificial light, the light use efficiency was about twice that of daylight. The starch concentration of the algae was unaffected by the light level and CO₂ concentration in the laboratory experiments (2.5–4.0 % of the dry weight). The flue gas concentration had no effect on starch concentration, while the starch concentration increased from about 1.5 % to about 6.0 % when the light source changed from artificial light to daylight. The flue gas from the silicomanganese smelter was characterized by a high CO₂ concentration (about 17 % v/v), low oxygen concentration (about 4 %), about 100 ppm NO _x , and 1 ppm SO₂. The biomass production using flue gas significantly increased as compared with about 5 % pure CO₂ gas, which was similar to the biomass produced at a CO₂ concentration of 10–20 % mixed with N₂. Thus, the enhanced biomass production seemed to be related to the low oxygen concentration rather than to the very high CO₂ concentration.     

Flue gas compounds and microalgae: (Bio-)chemical interactions leading to biotechnological opportunities

Flue gases are a resource yet to be fully utilised in microalgal biotechnology, not only to moderate the anthropogenic effects on our climate, but also to steer microalgal resource management towards innovative applications of microalgal biomass compounds. These gases, both untreated and treated into current discharge standards, contain CO₂, N₂, H₂O, O₂, NO_x, SO_x, C_xH_y, CO, particulate matter, halogen acids and heavy metals. To better steer and engineer flue gas-fed microalgal cultures, all these compounds need to be considered. Therefore, here, we review (i) the chemical composition and treatment technologies of flue gas, (ii) the uptake pathways and removal of the different compounds in microalgae reactors, and (iii) the tolerance and effects on microalgae of all flue gas compounds. By emphasising the interactions between microalgae and flue gas compounds, we envisage new pathways for microalgal biomass valorisation such as enzyme production for environmental technology, novel biogas production and biosequestration of minerals. Furthermore, we highlight fundamental and applied research niches that merit further investigation.     

Succinic acid production by <Emphasis Type="Italic">Actinobacillus succinogenes</Emphasis> from batch fermentation of mixed sugars

Succinic acid production from the monosaccharides xylose, arabinose, glucose, mannose and galactose was studied using the bacterium Actinobacillus succinogenes. In Duran bottle cultures, containing 10 g/L of each of sugar, succinic acid was produced from all sugars except for galactose. The highest succinate yield, 0.56 g/g, was obtained with glucose, whereas the succinate yield was 0.42, 0.38 and 0.44 g/g for xylose, mannose and arabinose, respectively. The specific succinate productivity was 0.7 g/g h for glucose, but below 0.2 g/g h for the other sugars. Batch bioreactor fermentations were carried out using a sugar mixture of the five sugars giving a total concentration of 50 g/L, mimicking the distribution of sugars in spent sulfite liquor (SSL) from Eucalyptus which is rich in xylose. In this mixture, an almost complete conversion of all sugars (except galactose) was achieved resulting in a final succinate concentration of 21.8–26.8 g/L and a total yield of 0.59–0.68 g/g. There was evidence of co-consumption of glucose and xylose, whereas mannose was consumed after glucose. The main by-products were acetate 0.14–0.20 g/g and formate 0.08–0.13 g/g. NADH balance calculations suggested that NADH required for succinate production was not met solely from formate and acetate production, but other means of NADH production was necessary. Results from mixed sugar fermentations were verified using SSL as substrate resulting in a succinate yield of 0.60 g/g. In addition, it was found that CO₂ sparging could replace carbonate supply in the form of MgCO₃ without affecting the succinate yield.     

Biogeochemical regime shifts in coastal landscapes: the contrasting effects of saltwater incursion and agricultural pollution on greenhouse gas emissions from a freshwater wetland

Many coastal plain wetlands receive nutrient pollution from agricultural fields and are particularly vulnerable to saltwater incursion. Although wetlands are a major source of the greenhouse gases methane (CH₄) and nitrous oxide (N₂O), the consequences of salinization for greenhouse gas emissions from wetlands with high agricultural pollution loads is rarely considered. Here, we asked how saltwater exposure alters greenhouse gas emissions from a restored freshwater wetland that receives nutrient loading from upstream farms. During March to November 2012, we measured greenhouse gases along a ~2 km inundated portion of the wetland. Sampling locations spanned a wide chemical gradient from sites receiving seasonal fertilizer nitrogen and sulfate (SO₄ ^2?) loads to sites receiving seasonal increases in marine salts. Concentrations and fluxes of CH₄ were low (<100 µg L^?1 and <10 mg m^?2 h^?1) for all sites and sampling dates when SO₄ ^2? was high (>10 mg L^?1), regardless of whether the SO₄ ^2? source was agriculture or saltwater. Elevated CH₄ (as high as 1,500 µg L^?1 and 45 mg m^?2 h^?1) was only observed on dates when air temperatures were >27 °C and SO₄ ^2? was <10 mg L^?1. Despite elevated ammonium (NH₄ ⁺) for saltwater exposed sites, concentrations of N₂O remained low (<5 µg L^?1 and <10 µg m^?2 h^?1), except when fertilizer derived nitrate (NO₃ ^?) concentrations were high and N₂O increased as high as 156 µg L^?1. Our results suggest that although both saltwater and agriculture derived SO₄ ^2? may suppress CH₄, increases in N₂O associated with fertilizer derived NO₃ ^? may offset that reduction in wetlands exposed to both agricultural runoff and saltwater incursion.     

Carbon and nitrogen mass balance during flue gas treatment with Dunaliella salina cultures

The biotreatment of flue gases with algae cultures is a promising option to sequestrate CO₂, yet the emission of other greenhouse gases (GHG) from the cultures can hamper their environmental benefit. Quantitative data on the sequestration potential for CO₂ and NO _x in relation to the direct production of CH₄ and N₂O are urgently required. The present study assessed the flows of carbon (C) and nitrogen (N) through cultures of the green alga Dunaliella salina, supplied with biodiesel flue gas, by means of mass balancing. D. salina was grown in artificially lighted, field- (42-L bubble column reactor) and laboratory-scale cultures (23 °C, pH 7.5). In the bubble column reactor, algae grew with an average specific growth rate of 0.237 day^?1 under flue gas supplementation (6.3 % (v/v) CO₂, 1.2 ppmv NO _x ), and CO₂ was retained to 39 % in the system. The specific sequestration rate for CO₂ was low, with 0.13 g CO₂ L^?1 day^?1. Cultures emitted up to 13.03 μg CH₄ L^?1 day^?1 and 4261 μg N₂O L^?1 day^?1. The moderate retention of NO _x -N was outweighed by emissions of N₂O-N, and total N in the system decreased by 15.48 % during the 9-day trial. Results suggest that GHG production was mainly the outcome of anaerobic microbial processes and their emission was lower in pre-sterilized cultures. Under the tested conditions, up to six times more CO₂ equivalents were emitted during flue gas treatment. Therefore, the direct GHG emissions of algae culture systems, intended for flue gas treatment (i.e. open ponds) need to be reviewed critically.     

Toxicity of flue gas components from cement plants in microalgae CO2 mitigation systems

Microalgae-based CO₂ capture from flue gas is an attractive mitigation strategy in the cement industry. However, NO _x and SO _x components might be harmful to microalgae. We performed toxicity assays, under 2 % (v/v) CO₂ and using nitrite, sulfite, or bisulfite salts, on an environmental isolate, identified as Desmodesmus abundans (The University of Texas at Austin (UTEX), no. 2976) and Scenedesmus sp. UTEX1589. Nitrite and sulfite did not inhibit growth at the tested concentrations (0–1,067 ppm (w/v) NO₂ ^? and 0–254 ppm (w/v) SO₃ ^2?); however, bisulfite was toxic above 39 ppm. Non-toxic concentrations of both sulfur-based compounds stimulated growth, but significantly higher growth rates were only observed for HSO₃ ^?. Within a narrower range, NO _x and SO _x served as a sole nutrient source. Overall, biomass production and growth rates of the environmental isolate were greater. A novel strategy to buffer high concentrations of HSO₃ ^? (200 ppm) was developed by adding cement kiln dust (CKD), a byproduct and flue gas component. The results suggest that CKD also provided other beneficial growth components and that sulfur optimization of the culture medium significantly increased carbon assimilation, particularly in D. abundans. In additional simulations of typical flue gas conditions in a modern cement plant (320, 40, and 40 ppm (w/v) of NO₂ ^?, SO₃ ^2?, and HSO₃ ^?, respectively, and 25 % (v/v) CO₂), along with the incorporation of 300 ppm CDK, growth of D. abundans was supported. Although further studies are needed, direct utilization of flue gas might be possible with the environmental isolate, where NO _x , SO _x , and CKD are all beneficial components of the mitigation system.     

Succinic acid production from corn stalk hydrolysate in an E. coli mutant generated by atmospheric and room-temperature plasmas and metabolic evolution strategies

AFP111 is a spontaneous mutant of Escherichia coli with mutations in the glucose-specific phosphotransferase system, pyruvate formate lyase system, and fermentative lactate dehydrogenase system, created to reduce byproduct formation and increase succinic acid accumulation. In AFP111, conversion of xylose to succinic acid only generates 1.67 ATP per xylose, but requires 2.67 ATP for xylose metabolism. Therefore, the ATP produced is not adequate to accomplish the conversion of xylose to succinic acid in chemically defined medium. An E. coli mutant was obtained by atmospheric and room-temperature plasmas and metabolic evolution strategies, which had the ability to use xylose and improve the capacity of cell growth. The concentration of ATP in the mutant was 1.33-fold higher than that in AFP111 during xylose fermentation. In addition, under anaerobic fermentation with almost 80 % xylose from corn stalk hydrolysate, a succinic acid concentration of 21.1 g l^?1 was obtained, with a corresponding yield of 76 %.     

Redirecting carbon flux through <Emphasis Type="Italic">pgi</Emphasis>-deficient and heterologous transhydrogenase toward efficient succinate production in <Emphasis Type="Italic">Corynebacterium glutamicum</Emphasis>

Corynebacterium glutamicum is particularly known for its potentiality in succinate production. We engineered C. glutamicum for the production of succinate. To enhance C₃–C₄ carboxylation efficiency, chromosomal integration of the pyruvate carboxylase gene pyc resulted in strain NC-4. To increase intracellular NADH pools, the pntAB gene from Escherichia coli, encoding for transhydrogenase, was chromosomally integrated into NC-4, leading to strain NC-5. Furthermore, we deleted pgi gene in strain NC-5 to redirect carbon flux to the pentose phosphate pathway (PPP). To solve the drastic reduction of PTS-mediated glucose uptake, the ptsG gene from C. glutamicum, encoding for the glucose-specific transporter, was chromosomally integrated into pgi-deficient strain resulted in strain NC-6. In anaerobic batch fermentation, the production of succinate in pntAB-overexpressing strain NC-5 increased by 14% and a product yield of 1.22 mol/mol was obtained. In anaerobic fed-batch process, succinic acid concentration reached 856 mM by NC-6. The yields of succinate from glucose were 1.37 mol/mol accompanied by a very low level of by-products. Activating PPP and transhydrogenase in combination led to a succinate yield of 1.37 mol/mol, suggesting that they exhibited a synergistic effect for improving succinate yield.     

High yield production of four-carbon dicarboxylic acids by metabolically engineered <Emphasis Type="Italic">Escherichia coli</Emphasis>

Several metabolic engineered Escherichia coli strains were constructed and evaluated for four-carbon dicarboxylic acid production. Fumarase A, fumarase B and fumarase C single, double and triple mutants were constructed in a ldhA adhE mutant background overexpressing the pyruvate carboxylase from Lactococcus lactis. All the mutants produced succinate as the main four-carbon (C4) dicarboxylic acid product when glucose was used as carbon source with the exception of the fumAC and the triple fumB fumAC deletion strains, where malate was the main C4-product with a yield of 0.61–0.67 mol (mole glucose)^?1. Additionally, a mdh mutant strain and a previously engineered high-succinate-producing strain (SBS550MG-Cms pHL413-Km) were investigated for aerobic malate production from succinate. These strains produced 40.38 mM (5.41 g/L) and 50.34 mM (6.75 g/L) malate with a molar yield of 0.53 and 0.55 mol (mole succinate)^?1, respectively. Finally, by exploiting the high-succinate production capability, the strain SBS550MG-Cms243 pHL413-Km showed significant malate production in a two-stage process from glucose. This strain produced 133 mM (17.83 g/L) malate in 47 h, with a high yield of 1.3 mol (mole glucose)^?1 and productivity of 0.38 g L^?1 h^?1.     

Withanolide A production from Withania somnifera hairy root cultures with improved growth by altering the concentrations of macro elements and nitrogen source in the medium

Withania somnifera is an important medicinal plant that contains withanolides as bioactive compounds. We have investigated the effects of macroelements and nitrogen source in hairy roots of W. somnifera with the aim of optimizing the production of biomass and withanolide A content. The effects of the macroelements NH₄NO₃, KNO₃, CaCl₂, MgSO₄ and KH₂PO₄ at concentrations of 0, 0.5, 1.0, 1.5 and 2.0× strengths and of nitrogen source [NH₄ ⁺/NO₃ ^? (0.00/18.80, 7.19/18.80, 14.38/18.80, 21.57/18.80, 28.75/18.80, 14.38/0.00, 14.38/9.40, 14.38/18.80, 14.38/28.20 and 14.38/37.60 mM)] in Murashige and Skoog medium were evaluated for biomass and withanolide A production. The highest accumulation of biomass (139.42 g l^?1 FW and 13.11 g l^?1 DW) was recorded in the medium with 2.0× concentration of KH₂PO₄, and the highest production of withanolide A was recorded with 2.0× KNO₃ (15.27 mg g^?1 DW). The NH₄ ⁺/NO₃ ^? ratio also influenced root growth and withanolide A production, with both parameters being larger when the NO₃ ^? concentration was higher than that of NH₄ ⁺. Maximum biomass growth (148.17 g l^?1 FW and 14.79 g l^?1 DW) was achieved at NH₄ ⁺/NO₃ ^? ratio of 14.38/37.60 mM, while withanolide A production was greatest (14.68 mg g^?1 DW) when the NH₄ ⁺/NO₃ ^? ratio was 0.00/18.80 mM. The results are useful for the large scale cultivation of Withania hairy root culture for the production of withanolide A.     

The citric acid production from raw glycerol by Yarrowia lipolytica yeast and its regulation

The optimal cultivation conditions ensuring the maximal rate of citric acid (CA) biosynthesis by glycerol-grown mutant Yarrowia lipolytica NG40/UV7 were found to be as follows: growth limitation by inorganic nutrients (nitrogen, phosphorus, or sulfur), 28 °C, pH 5.0, dissolved oxygen concentration (pO₂) of 50 % (of air saturation), and pulsed addition of glycerol from 20 to 80 g L^?1 depending on the rate of medium titration. Under optimal conditions of fed-batch cultivation, in the medium with pure glycerol, strain Y. lipolytica NG40/UV7 produced 115 g L^?1 of CA with the mass yield coefficient of 0.64 g g^?1 and isocitric acid (ICA) amounted to 4.6 g L^?1; in the medium with raw glycerol, CA production was 112 g L^?1 with the mass yield coefficient of 0.90 g g^?1 and ICA amounted to 5.3 g L^?1. Based on the activities of enzymes involved in the initial stages of raw glycerol assimilation, the tricarboxylic acid cycle and the glyoxylate cycle, the mechanism of increased CA yield from glycerol-containing substrates in Y. lipolytica yeast was explained.     

Alkaline-sulfite pretreatment and use of surfactants during enzymatic hydrolysis to enhance ethanol production from sugarcane bagasse

Sugarcane bagasse is a by-product from the sugar and ethanol industry which contains approximately 70 % of its dry mass composed by polysaccharides. To convert these polysaccharides into fuel ethanol it is necessary a pretreatment step to increase the enzymatic digestibility of the recalcitrant raw material. In this work, sugarcane bagasse was pretreated by an alkaline-sulfite chemithermomechanical process for increasing its enzymatic digestibility. Na₂SO₃ and NaOH ratios were fixed at 2:1, and three increasing chemical loads, varying from 4 to 8 % m/m Na₂SO₃, were used to prepare the pretreated materials. The increase in the alkaline-sulfite load decreased the lignin content in the pretreated material up to 35.5 % at the highest chemical load. The pretreated samples presented enhanced glucose yields during enzymatic hydrolysis as a function of the pretreatment severity. The maximum glucose yield (64 %) was observed for the samples pretreated with the highest chemical load. The use of 2.5 g l^?1 Tween 20 in the hydrolysis step further increased the glucose yield to 75 %. Semi-simultaneous hydrolysis and fermentation of the pretreated materials indicated that the ethanol yield was also enhanced as a function of the pretreatment severity. The maximum ethanol yield was 56 ± 2 % for the sample pretreated with the highest chemical load. For the sample pretreated with the lowest chemical load (2 % m/m NaOH and 4 % m/m Na₂SO₃), adding Tween 20 during the hydrolysis process increased the ethanol yield from 25 ± 3 to 39.5 ± 1 %.     

Effects of Growth Mode and Pyruvate Carboxylase on Succinic Acid Production by Metabolically Engineered Strains of Escherichia coli

Escherichia coli NZN111, which lacks activities for pyruvate-formate lyase and lactate dehydrogenase, and AFP111, a derivative which contains an additional mutation in ptsG (a gene encoding an enzyme of the glucose phophotransferase system), accumulate significant levels of succinic acid (succinate) under anaerobic conditions. Plasmid pTrc99A-pyc, which expresses the Rhizobium etli pyruvate carboxylase enzyme, was introduced into both strains. We compared growth, substrate consumption, product formation, and activities of seven key enzymes (acetate kinase, fumarate reductase, glucokinase, isocitrate dehydrogenase, isocitrate lyase, phosphoenolpyruvate carboxylase, and pyruvate carboxylase) from glucose for NZN111, NZN111/pTrc99A-pyc, AFP111, and AFP111/pTrc99A-pyc under both exclusively anaerobic and dual-phase conditions (an aerobic growth phase followed by an anaerobic production phase). The highest succinate mass yield was attained with AFP111/pTrc99A-pyc under dual-phase conditions with low pyruvate carboxylase activity. Dual-phase conditions led to significant isocitrate lyase activity in both NZN111 and AFP111, while under exclusively anaerobic conditions, an absence of isocitrate lyase activity resulted in significant pyruvate accumulation. Enzyme assays indicated that under dual-phase conditions, carbon flows not only through the reductive arm of the tricarboxylic acid cycle for succinate generation but also through the glyoxylate shunt and thus provides the cells with metabolic flexibility in the formation of succinate. Significant glucokinase activity in AFP111 compared to NZN111 similarly permits increased metabolic flexibility of AFP111. The differences between the strains and the benefit of pyruvate carboxylase under both exclusively anaerobic and dual-phase conditions are discussed in light of the cellular constraint for a redox balance.     

Succinate is the controller of O<Stack><Subscript>2</Subscript><Superscript>−</Superscript></Stack>/H<Subscript>2</Subscript>O<Subscript>2</Subscript> release at mitochondrial complex I : negative modulation by malate,positive by cyanide

Mitochondrial production of H₂O₂ is low with NAD substrates (glutamate/pyruvate, 3 and 2 mM) (G/P) and increases over ten times upon further addition of succinate, with the formation of a sigmoidal curve (semimaximal value at 290 μM, maximal H₂O₂ production at 600 μM succinate). Malate counteracts rapidly the succinate induced increased H₂O₂ release and moves the succinate dependent H₂O₂ production curve to the right. Nitric oxide (NO) and carbon monoxide (CO) are cytochrome c oxidase inhibitors which increase mitochondrial ROS production. Cyanide (CN⁻) was used to mimic NO and CO. In the presence of G/P and succinate (300 μM), CN⁻ progressively increased the H₂O₂ release rate, starting at 1.5 μM. The succinate dependent H₂O₂ production curve was moved to the left by 30 μM CN⁻. The V_max was little modified. We conclude that succinate is the controller of mitochondrial H₂O₂ production, modulated by malate and CN⁻. We propose that succinate promotes an interaction between Complex II and Complex I, which activates O₂⁻ production.     

Carbon dioxide utilisation of Dunaliella tertiolecta for carbon bio-mitigation in a semicontinuous photobioreactor

Bio-fixation of carbon dioxide (CO₂) by microalgae has been recognised as an attractive approach to offset anthropogenic emissions. Biological carbon mitigation is the process whereby autotrophic organisms, such as microalgae, convert CO₂ into organic carbon and O₂ through photosynthesis; this process through respiration produces biomass. In this study Dunaliella tertiolecta was cultivated in a semicontinuous culture to investigate the carbon mitigation rate of the system. The algae were produced in 1.2-L Roux bottles with a working volume of 1 L while semicontinuous production commenced on day 4 of cultivation when the carbon mitigation rate was found to be at a maximum for D. tertiolecta. The reduction in CO₂ between input and output gases was monitored to predict carbon fixation rates while biomass production and microalgal carbon content are used to calculate the actual carbon mitigation potential of D. tertiolecta. A renewal rate of 45 % of flask volume was utilised to maintain the culture in exponential growth with an average daily productivity of 0.07 g L^?1 day^?1. The results showed that 0.74 g L^?1 of biomass could be achieved after 7 days of semicontinuous production while a total carbon mitigation of 0.37 g L^?1 was achieved. This represented an increase of 0.18 g L^?1 in carbon mitigation rate compared to batch production of D. tertiolecta over the same cultivation period.     

High efficient conversion of CO2-rich bio-syngas to CO-rich bio-syngas using biomass char: a useful approach for production of bio-methanol from bio-oil

A novel approach for high efficient conversion of the CO₂-rich bio-syngas into the CO-rich bio-syngas was carried out by using biomass char and Ni/Al₂O₃ catalyst, which was successfully applied for production of bio-methanol from bio-oil. After the bio-syngas conditioning, the CO₂/CO ratio prominently dropped from 6.33 to 0.01-0.28. The maximum CO yield in the bio-syngas conditioning process reached about 1.96 mol/(mol CO₂) with a nearly complete conversion of CO₂ (99.5%). The performance of bio-methanol synthesis was significantly improved via the conditioned bio-syngas, giving a maximum methanol yield of 1.32 kg/(kg_catalyst h) with a methanol selectivity of 99%. Main reaction paths involved in the bio-syngas conditioning process have been investigated in detail by using different model mixture gases and different carbon sources.     

Key parameters for outdoor biomass production of Scenedesmus obliquus in solar tracked photobioreactors

The biomass productivity of Scenedesmus obliquus was investigated outdoors during all seasons in solar tracked flat panel photobioreactors (PBR) to evaluate key parameters for process optimization. CO₂ was supplied by flue gas from an attached combined block heat and power plant. Waste heat from the power plant was used to heat the culture during winter. The parameters pH, CO₂, and inorganic salt concentrations were automatically adjusted to nonlimiting levels. The optimum biomass concentration increased directly with the photosynthetic active radiation (PAR) from 3 to 5 g dry weight (DW)?L^?1 for a low PAR of 10 mol photons m^?2 day^?1 and high PAR of 40–60 mol photons m^?2 day^?1, respectively. The annual average biomass yield (photosynthetic efficiency) was 0.4?±?0.5 g DW mol^?1 photons. However, biomass yields of 1.5 g DW mol^?1 photons close to the theoretical maximum were obtained at low PAR. The productivity (including the night biomass losses) ranged during all seasons from ?5 up to 30 g DW m^?2 day^?1 with a mean productivity of 9?±?7 g DW m^?2 day^?1. Low night temperatures of the culture medium and elevated day temperatures to the species-specific optimum increased the productivity. Thus, continuous regulation of the biomass concentration and the culture temperature with regard to the fluctuating weather conditions is essential for process optimization of outdoor microalgal production systems in temperate climates.     

Microalgal removal of CO<Subscript>2</Subscript> from flue gases: Changes in medium pH and flue gas composition do not appear to affect the photochemical yield of microalgal cultures

Our research objectives are to determine under what conditions microalgal-based CO₂ capture from flue gases is economically attractive. Specifically, our objective here was to select microalgae that are temperature, pH and flue gas tolerant. Microalgae were grown under five different temperatures, three different pH and five different flue gas mixtures besides 100% CO₂ (gas concentrations that the cells were exposed to ranged 5.7–100% CO₂, 0–3504 ppm SO₂, 0–328 ppm NO, and 0–126 ppm NO₂). Our results indicate that the microalgal strains tested exhibit a substantial ability to withstand a wide range of temperature (54 strains tested), pH (20 strains tested) and flue gas composition (24 strains tested) likely to be encountered in cultures used for carbon sequestration from smoke stack gases. Our results indicate that microalgal photosynthesis is a limited but viable strategy for CO₂ capture from flue gases produced by stationary combustion sources.     

IMPROVED Candida methylica FORMATE DEHYDROGENASE FERMENTATION THROUGH STATISTICAL OPTIMIZATION OF LOW-COST CULTURE MEDIA

NAD⁺-dependent formate dehydrogenase (FDH, EC 1.2.1.2) is of use in the regeneration of NAD(P)H coenzymes, and therefore has strong potential for practical application in chemical and medical industries. A low-cost production of recombinant Escherichia coli (E. coli) containing FDH from Candida methylica (cmFDH) was optimized in molasses-based medium by using response surface methodology (RSM) based on central composite design (CCD). The beet molasses as a sole carbon source, (NH₄)₂HPO₄ as a nitrogen and phosphorus source, KH₂PO₄ as a buffer agent, and Mg₂SO₄ · 7H₂O as a magnesium and sulfur source were used as variables in the medium. The optimum medium composition was found to be 34.694 g L^?1 of reducing sugar (equivalent to molasses solution), 8.536 g L^?1 of (NH₄)₂HPO₄, 3.073 g L^?1 of KH₂PO₄, and 1.707 g L^?1 of Mg₂SO₄ · 7H₂O. Molasses-based culture medium increased the yield of cmFDH about three times compared to LB medium. The currently developed media has the potential to be used in industrial bioprocesses with low-cost production.     

Comparison of agarophytes (Gelidium, Gracilaria, and Gracilariopsis) as potential resources for bioethanol production

This study explores the possibility of producing ethanol using the acid hydrolysate of three abundant agar-containing red seaweeds (agarophytes): Gelidium amansii, Gracilaria tenuistipitata, and Gracilariopsis chorda. The main component in the seaweed samples was agar, which ranged from 20 to 51 % (g g^?1 dry weight). After optimizing acid hydrolysis, 100 g of seaweed was hydrolyzed at 130 °C for 15 min with 0.2 M H₂SO₄. Then, 120 mL of a 1:2 mixture of the hydrolysate broth and basal medium was fermented in a 200-mL bottle at 30 °C for 96 h. Of the three seaweeds, G. amansii had the best ethanol yield, producing 0.23 g g^?1 of galactose or 45 % of the theoretical yield. This yield increased to 60 % after detoxification of the hydrolysate with activated carbon.