Effect of nitrate feeding strategies on lipid and biomass productivities in fed-batch cultures of Nannochloropsis gaditana.

Controlled nitrate feeding strategies for fed-batch cultures of microalgae were applied for the enhancement of lipid production and microalgal growth rates. In particular, in this study, the effect of nitrate feeding rates on lipid and biomass productivities in fed-batch cultures of Nannochloropsis gaditana were investigated using three feeding modes (i.e., pulse, continuous, and staged) and under two light variations on both lipid productivity and fatty acid compositions. Higher nitrate levels negatively affected lipid production in the study. Increasing the light intensity increased the lipid contents of the microalgae in all three fed-batch feeding modes. A maximum of 58.3% lipid- to dry weight ratio was achieved when using pulse-fed cultures at an illumination of 200 µmol photons m-2  s-1 and 10 mg/day of nitrate feeding. This condition also resulted in the maximum lipid productivity of 44.6 mg L-1  day-1 . The fatty acid compositions of the lipids consisted predominantly of long-chain fatty acids (C:16 and C:18) and accounted for 70% of the overall fatty acid methyl esters. Pulse feeding mode was found to significantly enhance the biomass and lipid production. The other two feeding modes (continuous and staged) were not ideal for lipid and biomass production. This study demonstrates the applicability of pulse feeding strategies in fed-batch cultures as an appropriate cultivation strategy that can increase both lipid accumulation and biomass production.

An overview of microbial fuel cell usage in wastewater treatment, resource recovery and energy production.

Wastewater treatment is a high-cost and energy-intensive process not only due to large amounts of pollutants but also for the large volumes of water to be treated, which are mainly generated by human activities and different industries. In this regard, biological wastewater treatments have become substitutes to the current technologies, owing to the improved treatment efficiency and added value. Microbial fuel cells (MFCs) as one of the promising biological treatments have arisen as a viable solution for chemical oxygen demand (COD) removal and electricity generation simultaneously. Therefore, in this article, the effects of various operating conditions on the COD removal and power production from MFCs are thoroughly discussed. In addition, the advantages and weaknesses of current MFCs technologies used for different types of wastewater are summarized. Finally, the technical barriers facing by MFCs operation and the economic feasibility of using MFCs for wastewater treatment are provided.

Investigation of phyco-remediation of road salt run-off with marine microalgae Nannochloropsis gaditana.

Phyco-remediation is an environmental-friendly method, which involves the application of beneficial microalgae to treat wastewater-containing pollutants for a diverse range of conditions. Several industrial processes generate hyper saline wastewater, which is a significant challenge for conventional wastewater treatment, and the disposal of saline waters also has a negative impact on the environment. Road salt run-off is one such saline wastewater stream not currently treated and one that contributes significantly to negatively impacting receiving bodies of water. In this study, Nannochloropsis microalgae were able to assimilate >95% of the nitrates within 8 days in road salt concentrations ranging from 2.6% to 4.4% under phototrophic cultivation mode. Biomass yields of 1-2 g/l of culture were obtained with the maximum lipid of 22% (g/g) biomass in the road salt media. The crude road salt media provided all the essential micronutrients needed for algal cultivation. The fatty acid composition analysis of the obtained lipid composed of C16 and C18 over 45% of FAME are suitable for biofuel. This study has established that the use of road salt containing nitrate and phosphate nutrients will support the growth of marine micro algae for remediation of a waste water system that are the concern at winter-prevalent regions.

Process analysis and modeling of a single-step lutein extraction method for wet microalgae.

Lutein is a commercial carotenoid with potential health benefits. Microalgae are alternative sources for the lutein production in comparison to conventional approaches using marigold flowers. In this study, a process analysis of a single-step simultaneous extraction, saponification, and primary purification process for free lutein production from wet microalgae biomass was carried out. The feasibility of binary solvent mixtures for wet biomass extraction was successfully demonstrated, and the extraction kinetics of lutein from chloroplast in microalgae were first evaluated. The effects of types of organic solvent, solvent polarity, cell disruption method, and alkali and solvent usage on lutein yields were examined. A mathematical model based on Fick's second law of diffusion was applied to model the experimental data. The mass transfer coefficients were used to estimate the extraction rates. The extraction rate was found more significantly related with alkali ratio to solvent than to biomass. The best conditions for extraction efficiency were found to be pre-treatment with ultrasonication at 0.5 s working cycle per second, react 0.5 h in 0.27 L/g solvent to biomass ratio, and 1:3 ether/ethanol (v/v) with 1.25 g KOH/L. The entire process can be controlled within 1 h and yield over 8 mg/g lutein, which is more economical for scale-up.

Investigation of Chlorella vulgaris UTEX 265 Cultivation under Light and Low Temperature Stressed Conditions for Lutein Production in Flasks and the Coiled Tree Photo-Bioreactor (CTPBR).

Lutein has an increasing share in the pharmaceutical and nutraceutical market due to its benefits to eye health. Microalgae may be a potential source for lutein production while the expense limits the commercialization. In this study, a coiled tubular tree photobioreactor (CTPBR) design was investigated for cultivating the cold tolerant microalgae Chlorella vulgaris UTEX 265 under various conditions for lutein production. The influence and interaction of light irradiance strength, lighting cycle, and temperature on microalgae and lutein production efficiency at low temperature range were also studied in flasks via response surface method (RSM). The results demonstrated that 14 h day-light, 120 µmol photons m-2 s-1, and 10 °C was the optimal condition for algae growth and lutein production at low temperature experimental ranges. C. vulgaris UTEX 265 showed good potential to produce lutein in cold weather, and the optimum lutein production was contrary to the specific lutein content but corresponds to the trend of optimum growth. Additionally, fast growth (µ = 1.50 day-1) and good lutein recovery (11.98 mg g-1 day-1) in CTPBR were also achieved at the low irradiance stress condition and the low temperature photo-inhibition conditions.

Investigation of aqueous phase recycling for improving bio-crude oil yield in hydrothermal liquefaction of algae.

In this study, the aqueous phase obtained from catalytic/non-catalytic hydrothermal liquefaction (HTL) of Chlorella vulgaris was recycled as the reaction medium with an aim to reduce water consumption and increase bio-crude oil yield. Although both Na2CO3 and HCOOH catalysts have been proven to be effective for promoting biomass conversion, the bio-crude oil yield obtained from HTL with Na2CO3 (11.5wt%) was lower than that obtained from the non-catalytic HTL in pure water at 275°C for 50min. While, the HCOOH led to almost the same bio-crude yield from HTL (29.4wt%). Interestingly, bio-crude oil yield obtained from non-catalytic or catalytic HTL in recycled aqueous phase was much higher than that obtained from HTL in pure water. Recycling aqueous phase obtained from catalytic HTL experiments resulted in a sharp increase in the bio-crude oil yield by 32.6wt% (Na2CO3-HTL) and 16.1wt% (HCOOH-HTL), respectively.

Carotenoids from microalgae: A review of recent developments.

Carotenoids have been receiving increasing attention due to their potential health benefits. Microalgae are recognized as a natural source of carotenoids and other beneficial byproducts. However, the production of micro-algal carotenoids is not yet sufficiently cost-effective to compete with traditional chemical synthetic methods and other technologies such as extraction from plant based sources. This review presents the recent biotechnological developments in microalgal carotenoid production. The current technologies involved in their bioprocessing including cultivation, harvesting, extraction, and purification are discussed with a specific focus on downstream processing. The recent advances in chemical and biochemical synthesis of carotenoids are also reviewed for a better understanding of suitable and economically feasible biotechnological strategies. Some possible future directions are also proposed.

Expression of exo-inulinase gene from Aspergillus niger 12 in E. coli strain Rosetta-gami B (DE3) and its characterization.

Inulin is a linear carbohydrate polymer of fructose subunits (2-60) with terminal glucose units, produced as carbon storage in selected plants. It cannot directly be taken up by most microorganisms due to its large size, unless prior hydrolysis through inulinase enzymes occurs. The hydrolyzed inulin can be taken up by microbes and/or recovered and used industrially for the production of high fructose syrup, inulo-oligosaccharides, biofuel, and nutraceuticals. Cell-free enzymatic hydrolysis would be desirable for industrial applications, hence the recombinant expression, purification and characterization of an Aspergillus niger derived exo-inulinase was investigated in this study. The eukaroyototic exo-inulinase of Aspergillus niger 12 has been expressed, for the first time, in an E. coli strain [Rosetta-gami B (DE3)]. The molecular weight of recombinant exo-inulinase was estimated to be ∼81 kDa. The values of Km and Vmax of the recombinant exo-inulinase toward inulin were 5.3 ± 1.1 mM and 402.1 ± 53.1 µmol min(-1)  mg(-1) protein, respectively. Towards sucrose the corresponding values were 12.20 ± 1.6 mM and 902.8 ± 40.2 µmol min(-1)  mg(-1) protein towards sucrose. The S/I ratio was 2.24 ± 0.7, which is in the range of native inulinase. The optimum temperature and pH of the recombinant exo-inulinase towards inulin was 55°C and 5.0, while they were 50°C and 5.5 towards sucrose. The recombinant exo-inulinase activity towards inulin was enhanced by Cu(2+) and reduced by Fe(2+) , while its activity towards sucrose was enhanced by Co(2+) and reduced by Zn(2+) . © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:629-637, 2016.

Investigation of high pressure steaming (HPS) as a thermal treatment for lipid extraction from Chlorella vulgaris.

Biofuels from algae are considered a technically viable energy source that overcomes several of the problems present in previous generations of biofuels. In this research high pressure steaming (HPS) was studied as a hydrothermal pre-treatment for extraction of lipids from Chlorella vulgaris, and analysis by response surface methodology allowed finding operational points in terms of target temperature and algae concentration for high lipid and glucose yields. Within the range covered by these experiments the best conditions for high bio-crude yield are temperatures higher than 174°C and low biomass concentrations (<5 g/L). For high glucose yield there are two suitable operational ranges, either low temperatures (<105°C) and low biomass concentrations (<4 g/L); or low temperatures (<105°C) and high biomass concentrations (<110 g/L). High pressure steaming is a good hydrothermal treatment for lipid recovery and does not significantly change the fatty acids profile for the range of temperatures studied.

Effect of biodiesel addition on microbial community structure in a simulated fuel storage system.

Understanding changes in microbial structure due to biodiesel storage is important both for protecting integrity of storage systems and fuel quality management. In this work a simulated storage system was used to study the effect of biodiesel (0%, 25%, 50%, 75% and 100%) on a microbial population, which was followed by community level physiological profiling (CLPP), 16s rDNA analysis and plating in selective media. Results proved that structure and functionality were affected by biodiesel. CLPP showed at least three populations: one corresponding to diesel, one to biodiesel and one to blends of diesel and biodiesel. Analysis of 16s rDNA revealed that microbial composition was different for populations growing in diesel and biodiesel. Genera identified are known for degradation of hydrocarbons and emulsifier production. Maximum growth was obtained in biodiesel; however, microbial counts in standard media were lower for this samples. Acidification of culture media was observed at high biodiesel concentration.

Investigation of biomass concentration, lipid production, and cellulose content in Chlorella vulgaris cultures using response surface methodology.

The microalgae Chlorella vulgaris produce lipids that after extraction from cells can be converted into biodiesel. However, these lipids cannot be efficiently extracted from cells due to the presence of the microalgae cell wall, which acts as a barrier for lipid removal when traditional extraction methods are employed. Therefore, a microalgae system with high lipid productivity and thinner cell walls could be more suitable for lipid production from microalgae. This study addresses the effect of culture conditions, specifically carbon dioxide and sodium nitrate concentrations, on biomass concentration and the ratio of lipid productivity/cellulose content. Optimization of culture conditions was done by response surface methodology. The empirical model for biomass concentration (R(2) = 96.0%) led to a predicted maximum of 1123.2 mg dw L(-1) when carbon dioxide and sodium nitrate concentrations were 2.33% (v/v) and 5.77 mM, respectively. For lipid productivity/cellulose content ratio (R(2) = 95.2%) the maximum predicted value was 0.46 (mg lipid L(-1) day(-1) )(mg cellulose mg biomass(-1) )(-1) when carbon dioxide concentration was 4.02% (v/v) and sodium nitrate concentration was 3.21 mM. A common optimum point for both variables (biomass concentration and lipid productivity/cellulose content ratio) was also found, predicting a biomass concentration of 1119.7 mg dw L(-1) and lipid productivity/cellulose content ratio of 0.44 (mg lipid L(-1) day(-1) )(mg cellulose mg biomass(-1) )(-1) for culture conditions of 3.77% (v/v) carbon dioxide and 4.01 mM sodium nitrate. The models were experimentally validated and results supported their accuracy. This study shows that it is possible to improve lipid productivity/cellulose content by manipulation of culture conditions, which may be applicable to any scale of bioreactors.

Engineering challenges in biodiesel production from microalgae.

In recent years, the not too distant exhaustion of fossil fuels is becoming apparent. Apart from this, the combustion of fossil fuels leads to environmental concerns, the emission of greenhouse gases and issues with global warming and health problems. Production of biodiesel from microalgae may represent an attractive solution to the above mentioned problems, and can offer a renewable source of fuel with fewer pollutants. This review presents a compilation of engineering challenges related to microalgae as a source of biodiesel. Advantages and current limitations for biodiesel production are discussed; some aspects of algae cells biology, with emphasis on cell wall composition, as it represents a barrier for fatty acid extraction and lipid droplets are also presented. In addition, recent advances in the different stages of the manufacturing process are included, starting from the strain selection and finishing in the processing of fatty acids into biodiesel.

Soy protein recovery in a solvent-free process using continuous liquid-solid circulating fluidized bed ion exchanger.

Soy protein concentrates and soy protein isolates act as ingredients in bakery, meat and dairy products, baby formulas, starting materials for spun textured vegetable products, and other nutritional supplements. In this study, the effectiveness of a liquid-solid circulating fluidized bed (LSCFB) ion exchanger is demonstrated for the recovery of soluble soy proteins from full fat and defatted soy flour. Under steady-state operating conditions, about 50% of the proteins could be recovered from the feed streams entering the ion exchanger. The LSCFB was shown to be a promising system for the recovery of soy protein from both defatted and full fat soy flour solutions. As the ion exchange process captures dissolved proteins, the system may offer a less damaging form of processing compared with the acid precipitation process where soy protein aggregates form and functionality is affected. In addition, the LSCFB allows simultaneous adsorption and desorption of the proteins allowing for a continuous operation. No prefiltration of feed containing suspended particles is required as well, because fluidization is used in place of packed bed technology to improve on current ion exchange processes.

Modeling and simulation of liquid-solid circulating fluidized bed ion exchange system for continuous protein recovery.

Liquid-solid circulating fluidized bed (LSCFB) is an integrated two-column (downcomer and riser) system which can accommodate two separate processes (adsorption and desorption) in the same unit with continuous circulation of the solid particles between the two columns. In this study, a mathematical model based on the assumption of homogeneous fluidization was developed considering hydrodynamics, adsorption-desorption kinetics and liquid-solid mass transfer. The simulation results showed good agreement with the available experimental results for continuous protein recovery. A parametric sensitivity study was performed to better understand the influence of different operating parameters on the BSA adsorption and desorption capacity of the system. The model developed can easily be extended to other applications of LSCFB.

Multiobjective optimization of the operation of a liquid-solid circulating fluidized bed ion-exchange system for continuous protein recovery.

Like most real-life processes, the operation of liquid-solid circulating fluidized bed (LSCFB) system for continuous protein recovery is associated with several objectives such as maximization of production rate and recovery of protein, and minimization of amount solid ion-exchange resin requirement, all of which need to be optimized simultaneously. In this article, multiobjective optimization of a LSCFB system for continuous protein recovery was carried out using an experimentally validated mathematical model to find the scope for further improvements in its operation. Elitist non-dominated sorting genetic algorithm with its jumping gene adaptation was used to solve a number of bi- and tri-objective function optimization problems. The optimization resulted in Pareto-optimal solution, which provides a broad range of non-dominated solutions due to conflicting behavior of the operating parameters on the system performance indicators. Significant improvements were achieved, for example, the production rate at optimal operation increased by 33%, using 11% less solid compared to reported experimental results for the same recovery level. The effects of operating variables on the optimal solutions are discussed in detail. The multiobjective optimization study reported here can be easily extended for the improvement of LSCFB system for other applications.

Bioelectronic imaging array based on bacteriorhodopsin film.

A photoreceptor array that exploits the light sensitive bacteriorhodopsin (bR) films has been manufactured on a flexible indium-tin-oxide (ITO) coated plastic film using electrophoretic sedimentation technique (EPS). The effective sensing area of each photoreceptor is 2 x 2 mm (2), separated by 1 mm and arranged in a 4 x 4 array. A switched integrator with gain on the order of 10(10) is used to amplify the signal to a suitable level. When exposed to light, the differential response characteristic is attributed to charge displacement and recombination within bR molecules, as well as loading effects of the attached amplifier. The peak spectral response occurs at 568 nm and is linear over the tested light power range of 200 mu W to 12 mW. The response remains linear at other tested wavelengths, but with reduced amplitude. Initial tests have indicated that responsivity among all photoreceptors is greater than 71% of the average value, 465.25 mV/mW. The differential nature of the signal generated by bR makes it a suitable sensing material for vision applications such as motion detection. The prototype array demonstrates this property by employing Reichardt's delay-and-correlate algorithm. Furthermore, fabricating sensor arrays on flexible substrates introduces a new design approach that enables non-planar imaging surfaces.

Investigation of a dual-particle liquid-solid circulating fluidized bed bioreactor for extractive fermentation of lactic acid.

A dual-particle liquid-solid circulating fluidized bed (DP-LSCFB) bioreactor has been constructed and investigated for the simultaneous production and extraction of lactic acid using immobilized Lactobacillus bulgaricus and ion-exchange resins. The apparatus consisted of a downer fluidized bed, 13 cm I.D. and 4.75 m tall, and a riser fluidized bed, 3.8 cm I.D. and 5.15 m in height. The lactic acid production and removal was carried out in the downer, while the riser was used for the recovery of lactic acid. A continuously recirculating bed of ion-exchange resin was used for adsorption of the produced acid as well as for maintaining optimum pH for bioconversion, thus eliminating the need for costly and complex chemical control approach used in conventional techniques. Studies using lactic acid aqueous solution as feed and sodium hydroxide solution as regeneration stream showed 93% lactic acid removal from the downer and 46% recovery in the riser under the conditions investigated. Such results prove the functionality of using the newly developed bioreactor design for the continuous production and recovery of products of biotechnological significance.

Fate of copper in submerged membrane bioreactors treating synthetic municipal wastewater.

This paper assesses the impact of copper on the performance of two membrane bioreactors (MBR) treating municipal wastewater at a hydraulic retention time (HRT) of 4h, and solids residence times (SRT) of 20 days, at influent copper concentrations of 0.2-8 mg Cu/L. The addition of copper resulted in a significant increase in soluble microbial products (SMPs), and a predominance of >100 kDa molecular weight SMPs. The study showed that in well-buffered wastewaters, complete nitrification was achieved at total copper concentrations as high as 840 mg/L or 10% of the mixed liquor volatile suspended solids. MINTEQ simulation showed that most of the copper (99.8%) in the MBR was in the form of inorganic copper precipitates, with free Cu2+ and total soluble copper in the range of 0.0-0.11 and 0.1-0.82 mg/L, respectively.

Biodesulfurization of refractory organic sulfur compounds in fossil fuels.

The stringent new regulations to lower sulfur content in fossil fuels require new economic and efficient methods for desulfurization of recalcitrant organic sulfur. Hydrodesulfurization of such compounds is very costly and requires high operating temperature and pressure. Biodesulfurization is a non-invasive approach that can specifically remove sulfur from refractory hydrocarbons under mild conditions and it can be potentially used in industrial desulfurization. Intensive research has been conducted in microbiology and molecular biology of the competent strains to increase their desulfurization activity; however, even the highest activity obtained is still insufficient to fulfill the industrial requirements. To improve the biodesulfurization efficiency, more work is needed in areas such as increasing specific desulfurization activity, hydrocarbon phase tolerance, sulfur removal at higher temperature, and isolating new strains for desulfurizing a broader range of sulfur compounds. This article comprehensively reviews and discusses key issues, advances and challenges for a competitive biodesulfurization process.