Industrial Photobioreactors Research Articles

Assessment of photosynthetic activity in dense microalgae cultures using oxygen production

Microalgae are photosynthetic microorganisms playing a pivotal role in primary production in aquatic ecosystems, sustaining the entry of carbon in the biosphere. Microalgae have also been recognized as sustainable source of biomass to complement crops. For this objective they are cultivated in photobioreactors or ponds at high cell density to maximize biomass productivity and lower the cost of downstream processes.Photosynthesis depends on light availability, that is often not constant over time. In nature, sunlight fluctuates over diurnal cycles and weather conditions. In high-density microalgae cultures of photobioreactors outdoors, on top of natural variations, microalgae are subjected to further complexity in light exposure. Because of the high-density cells experience self-shading effects that heavily limit light availability in most of the mass culture volume. This limitation strongly affects biomass productivity of industrial microalgae cultivation plants with important implications on economic feasibility.Understanding how photosynthesis responds to cell density is informative to assess functionality in the inhomogeneous light environment of industrial photobioreactors. In this work we exploited a high-sensitivity Clark electrode to measure microalgae photosynthesis and compare cultures with different densities, using Nannochloropsis as model organism. We observed that cell density has a substantial impact on photosynthetic activity, and demonstrated the reduction of the cell's light-absorption capacity by genetic modification is a valuable strategy to increase photosynthetic functionality on a chlorophyll-basis of dense microalgae cultures.

Efficient Production of Microalgal Biomass—Step by Step to Industrial Scale

The production of microalgal biomass on a commercial scale remains a significant challenge. Despite the positive results obtained in the laboratory, there are difficulties in obtaining similar results in industrial photobioreactors. Changing the cultivation conditions can affect not only the growth of microalgae but also their metabolism. This is of particular importance for the use of biomass for bioenergy production, including biofuel production. The aim of this study was to determine the biomass production efficiency of selected microalgal strains, depending on the capacity of the photobioreactor. The lipid and ash content of the biomass were also taken into account. It was found that as the scale of production increased, the amount of biomass decreased, irrespective of the type of strain. The change in scale also affected the lipid content of the biomass. The highest values were found in 2.5 L photobioreactors (ranging from 26.3 ± 2.2% for Monoraphidium to 13.9 ± 0.3% for Chlorella vulgaris). The least favourable conditions were found with industrial photobioreactors, where the lipid content of the microalgal biomass ranged from 7.1 ± 0.6% for Oocycstis submarina to 10.2 ± 1.2% for Chlorella fusca. The increase in photobioreactor capacity had a negative effect on the ash content.

Complete Genome Sequence of Phormidium yuhuli AB48, Isolated from an Industrial Photobioreactor Environment

We report the genome of Phormidium yuhuli AB48, which includes a circular chromosome and a circular plasmid (4,747,469 bp and 51,599 bp, respectively). This is currently the only closed reference genome of an isolate of the Phormidium genus, based on the Genome Taxonomy Database (GTDB), providing a potential model system for sustainable biotechnology innovation.

Metagenome-Assembled Genomes for "Candidatus Phormidium sp. Strain AB48" and Co-occurring Microorganisms from an Industrial Photobioreactor Environment.

Here, we report metagenome-assembled genomes for "Candidatus Phormidium sp. strain AB48" and three cooccurring microorganisms from a biofilm-forming industrial photobioreactor environment, using the PacBio sequencing platform. Several mobile genetic elements, including a double-stranded DNA phage and plasmids, were also recovered, with the potential to mediate gene transfer within the biofilm community.

Nutritional Composition and Untargeted Metabolomics Reveal the Potential of Tetradesmus obliquus, Chlorella vulgaris and Nannochloropsis oceanica as Valuable Nutrient Sources for Dogs.

Simple SummaryThe growing pet population is fuelling the debate over the environmental impact of the pet food sector and questioning its sustainability, unveiling the need for alternative and more sustainable ingredients. The present study relates the nutritional composition and metabolomic profile of the microalgae Tetradesmus obliquus, Chlorella vulgaris, and Nannochloropsis oceanica to the nutritional requirements of dogs to assess their potential as alternative foods. Overall, the essential amino acid content exceeded the amounts required for dogs at all life stages, except methionine and cysteine, and their different fatty acid and mineral profiles demonstrates a different potential for dog feeding. In addition, fiber was mainly composed of insoluble dietary fiber and untargeted metabolomics highlighted glycolipids, glycerolipids and phospholipids as the most discriminating compounds. The results support the potential of T. obliquus, C. vulgaris and N. oceanica as valuable nutrient sources for dogs.The growing pet population is questioning the sustainability of the pet food system. Although microalgae may constitute a more sustainable food resource, the assessment of their potential for canine diets is almost non-existent. The present study aimed to evaluate the potential of three microalgae species (Tetradesmus obliquus, Chlorella vulgaris and Nannochloropsis oceanica) grown locally in industrial photobioreactors as alternative food resources for dogs. A detailed characterization of their nutritional composition and metabolomic profile was carried out and related to the nutritional requirements of dogs. Overall, the essential amino acid content exceeded the amounts required for dogs at all life stages, except methionine and cysteine. The three microalgae were deficient in linoleic acid, N. oceanica presented a linolenic acid content below requirements and T. obliquus and C. vulgaris were deficient in arachidonic and eicosapentaenoic acids. The fiber was mainly composed of insoluble dietary fiber. The mineral profile varied greatly with the microalgae species, demonstrating their different potential for dog feeding. Untargeted metabolomics highlighted glycolipids, glycerolipids and phospholipids as the most discriminating compounds between microalgae species. Overall, the results support the potential of T. obliquus, C. vulgaris and N. oceanica as valuable macro- and micro-nutrients sources for dog feeding.

The survivor strain: isolation and characterization of Phormidium yuhuli AB48, a filamentous phototactic cyanobacterium with biotechnological potential.

Despite their recognized potential, current applications of cyanobacteria as microbial cell factories remain in early stages of development. This is partly due to the fact that engineered strains are often difficult to grow at scale. This technical challenge contrasts with the dense and highly productive cyanobacteria populations thriving in many natural environments. It has been proposed that the selection of strains pre-adapted for growth in industrial photobioreactors could enable more productive cultivation outcomes. Here, we described the initial morphological, physiological, and genomic characterization of Phormidium yuhuli AB48 isolated from an industrial photobioreactor environment. P. yuhuli AB48 is a filamentous phototactic cyanobacterium with a growth rate comparable to Synechocystis sp. PCC 6803. The isolate forms dense biofilms under high salinity and alkaline conditions and manifests a similar nutrient profile to Arthrospira platensis (Spirulina). We sequenced, assembled, and analyzed the P. yuhuli AB48 genome, the first closed circular isolate reference genome for a member of the Phormidium genus. We then used cultivation experiments in combination with proteomics and metabolomics to investigate growth characteristics and phenotypes related to industrial scale cultivation, including nitrogen and carbon utilization, salinity, and pH acclimation, as well as antibiotic resistance. These analyses provide insight into the biological mechanisms behind the desirable growth properties manifested by P. yuhuli AB48 and position it as a promising microbial cell factory for industrial-scale bioproduction[221, 1631].

Evaluation of an Interpolated Controller in an Industrial Photobioreactor

Nowadays, some of the most important challenges to society are focused on the use of renewable energies, mitigation of greenhouse-type gases and residual treatment. Microalgae are becoming one of the most powerful solutions to contribute to these problems. So, industrial facilities have been developed in the last decades to produce this type of microorganisms. However the dynamics of these systems is very complex, and advanced models and control strategies are required. The Interpolating Control is a novel technique with capacity to deal with state and control constraints with low computational burden, being suitable to be applied in this type of processes. This technique has been used in the literature to control simulated or laboratory scale systems. This paper shows the operation results for a Interpolated Controller in an industrial raceway photobioreactor to contribute in the control of these real processes.

Technological mapping and trends in photobioreactors for the production of microalgae.

Photobioreactors (PBRs) are equipment of central importance for the massive cultivation of microalgae, providing controlled conditions for high cell productivity. There are a few popular PBR designs, with contrasting advantages and limitations, such as poor light distribution, mass transfer, or hydrodynamic behavior. Due to the environmental concerns in recent decades and the discovery of new, useful microalgal metabolites, the interest in finding alternatives to solve technological bottlenecks of PBRs has intensified. In this process, new geometries, materials, and modes of light supply were developed, generating a significant scientific and technological output, reported in papers and patents. We present a technological landscape analysis of photobioreactor design, focusing on improvements of the classical geometries and trends in industrial photobioreactors. The analysis of 412 patent documents showed a surge in innovation filing since 2005 and a reduction in the number of new documents along the last decade. The recent efforts in design improvement, the leading countries, institutes and companies that innovate, and the trends in PBR technology are presented and discussed.

Nutritional Potential and Toxicological Evaluation of Tetraselmis sp. CTP4 Microalgal Biomass Produced in Industrial Photobioreactors.

Commercial production of microalgal biomass for food and feed is a recent worldwide trend. Although it is common to publish nutritional data for microalgae grown at the lab-scale, data about industrial strains cultivated in an industrial setting are scarce in the literature. Thus, here we present the nutritional composition and a microbiological and toxicological evaluation of Tetraselmis sp. CTP4 biomass, cultivated in 100-m3 photobioreactors at an industrial production facility (AlgaFarm). This microalga contained high amounts of protein (31.2 g/100 g), dietary fibres (24.6 g/100 g), digestible carbohydrates (18.1 g/100 g) and ashes (15.2 g/100 g), but low lipid content (7.04 g/100 g). The biomass displayed a balanced amount of essential amino acids, n-3 polyunsaturated fatty acids, and starch-like polysaccharides. Significant levels of chlorophyll (3.5 g/100 g), carotenoids (0.61 g/100 g), and vitamins (e.g., 79.2 mg ascorbic acid /100 g) were also found in the biomass. Conversely, pathogenic bacteria, heavy metals, cyanotoxins, mycotoxins, polycyclic aromatic hydrocarbons, and pesticides were absent. The biomass showed moderate antioxidant activity in several in vitro assays. Taken together, as the biomass produced has a balanced biochemical composition of macronutrients and (pro-)vitamins, lacking any toxic contaminants, these results suggest that this strain can be used for nutritional applications.

New insights into developing antibiofouling surfaces for industrial photobioreactors.

The biofouling formation of the marine microalga Nannochloropsis gaditana on nontoxic surfaces was quantified on rigid materials, both coated (with fouling release coatings and nanoparticle coatings) and noncoated, to cover a wide range of surface properties from strongly hydrophobic to markedly hydrophilic under conditions similar to those prevailing in outdoor massive cultures of marine microalgae. The effect of seawater on surfaces that presented the best antibiofouling properties was also evaluated. The adhesion intensity on the different surfaces was compared with the predictions of the biocompatibility theories developed by Baier and Vogler using water adhesion tension (τ0 ) as the quantitative parameter of surface wettability. For the most hydrophobic surfaces, τ0 ≤ 0, the microalgae adhesion density increased linearly with τ0 , following the Baier's theory trend. However, for the rest of the surfaces, τ0 ≥ 0, a tendency toward minimum adhesion was observed for amphiphilic surfaces with a τ0 = 36 mJ/m2 , a value close to that which minimizes cell adhesion according to Vogler's theory. The understanding and combination of the two biocompatibility theories could help to design universal antibiofouling surfaces that minimize the van der Waals forces and prevent foulant adsorption by using a thin layer of hydration.

A novel, nature-based alternative for photobioreactor decontaminations

Large-scale microalgae cultivations are increasingly used for the production of animal feed, nutritional supplements and various high-value bioproducts. Due to the process size and other limitations, contaminations of microalgae fermentations with other photoautotrophic microorganism are frequently observed. In the present study, we explored the applicability of 5-isobutyl-2,3-dimethylpyrazine for the removal of contaminating microalgae from industrial photobioreactors. In order to select a representative microbial population for susceptibility experiments, reactor samples were obtained from a multi-stage cultivation process. Assignments of 18S rRNA gene fragment amplicons indicated that Haematococcus, Chlorella, and Scenedesmus were the three most frequently occurring microalgae genera in the selected reactors. Following the isolation of representative algae cultures, susceptibility tests were conducted with the antimicrobial pyrazine. It was demonstrated that all isolated contaminants are highly susceptible to the bioactive compound. The highest tolerance towards the alkylpyrazine was observed with Scenedesmus vacuolatus; solutions with 1.66% (v/v) of the active compound were required for its deactivation. Further tests with the vaporized pyrazine showed consistent reductions in the viability of treated microalgae. This pilot study provides evidence for the applicability of a novel, nature-based alternative for bioreactor decontaminations.

Industrial production of Phaeodactylum tricornutum for CO2 mitigation: biomass productivity and photosynthetic efficiency using photobioreactors of different volumes

The photosynthetic efficiency (PE) and potential of Phaeodactylum tricornutum for CO2 mitigation in industrial tubular photobioreactors (PBRs) of different volumes were evaluated. A preliminary assay was performed at lab-scale to optimize the salt concentration of the culture medium. Interestingly, salinity did not affect the growth of P. tricornutum at concentrations of 2.5, 5, 10, and 20 g L−1. Higher volumetric productivities were achieved in the 2.5-m3 tubular PBR (0.235 g L−1 day−1), followed by 35- and 10-m3 PBRs. Maximum areal productivities corresponded to 48.5, 45.0, and 12.8 g m−2 day−1 for the 35-, 10-, and 2.5-m3 PBRs, respectively. PE was thus higher in the 35- and 10-m3 PBRs (2.21 and 2.08%, respectively). The 10- and 35-m3 PBR showed CO2 mitigation efficiencies of 60 and 41%, respectively, of the CO2 introduced into the PBR, corresponding to 2.3 and 2.5 g of fixed CO2 per g of biomass. Overall, cultivation of P. tricornutum couples high PE and areal productivity when the industrial PBRs were used, particularly PBRs of larger volumes. This improved PE performance with larger PBR volumes strongly suggests that large-scale cultivation of this diatom holds great potential for industrial CO2 mitigation.

Biomass estimation of an industrial raceway photobioreactor using an extended Kalman filter and a dynamic model for microalgae production

Production of microalgae is one of the emerging biotechnological processes due to its potential applications to produce high value-added compounds. In photobioreactors for microalgae production, the biomass concentration is a desirable variable to be measured on-line to optimize the yield of the systems. However, biomass concentration can hardly be monitored in real time. There are few expensive commercial sensors that in fact provide uncertain measurements. State estimators, also known as software sensors, are algorithms designed to estimate unmeasured (or non-easily measurable) variables of a process. In this work, a state estimator using the extended Kalman filter algorithm is developed to estimate biomass concentration for an outdoor industrial raceway photobioreactor. The state estimator is based on a dynamic model for microalgae production specifically designed for this type of photobioreactor. Results demonstrate that, despite the complex non-linear dynamics that characterise this kind of bioprocess, a state estimator can provide a relatively accurate estimation of the biomass concentration. Furthermore, a state estimator could be used to optimize the operation of industrial photobioreactors by utilizing the estimated biomass concentration for automatic control of the process.

Scale-up and large-scale production of Tetraselmis sp. CTP4 (Chlorophyta) for CO2 mitigation: from an agar plate to 100-m3 industrial photobioreactors

Industrial production of novel microalgal isolates is key to improving the current portfolio of available strains that are able to grow in large-scale production systems for different biotechnological applications, including carbon mitigation. In this context, Tetraselmis sp. CTP4 was successfully scaled up from an agar plate to 35- and 100-m3 industrial scale tubular photobioreactors (PBR). Growth was performed semi-continuously for 60 days in the autumn-winter season (17th October – 14th December). Optimisation of tubular PBR operations showed that improved productivities were obtained at a culture velocity of 0.65–1.35 m s−1 and a pH set-point for CO2 injection of 8.0. Highest volumetric (0.08 ± 0.01 g L−1 d−1) and areal (20.3 ± 3.2 g m−2 d−1) biomass productivities were attained in the 100-m3 PBR compared to those of the 35-m3 PBR (0.05 ± 0.02 g L−1 d−1 and 13.5 ± 4.3 g m−2 d−1, respectively). Lipid contents were similar in both PBRs (9–10% of ash free dry weight). CO2 sequestration was followed in the 100-m3 PBR, revealing a mean CO2 mitigation efficiency of 65% and a biomass to carbon ratio of 1.80. Tetraselmis sp. CTP4 is thus a robust candidate for industrial-scale production with promising biomass productivities and photosynthetic efficiencies up to 3.5% of total solar irradiance.

Hydrodynamic and mass transfer characterization of flat-panel airlift photobioreactors for the cultivation of a photosynthetic microbial consortium

The design and build of industrial photobioreactors are critical steps for the production of photoautotrophic microorganisms. Flat-panel airlift photobioreactors (PBRs) of 50L and 150L were built; hydrodynamics (gas hold-up (ε), mixing time (tm) and Reynolds number (Re)) and mass transfer properties were characterized in the two flat-panel airlift PBRs (50 and 150L). The geometric ratio (illuminated area: operation volume (A/V) ratio) selected for building the 150L PBR was adequate to carry out biomass production of a microbial consortium (MC). A high CO2 utilization efficiency in the PBRs was obtained. Finally, the experimental results of kLa were correlated mathematically to describe the influences of the considered factors (the overall power input and density) on the studied superficial gas velocity range. The predicted kLa values yielded acceptable results compared to the experimental results.

Gyrotactic swimmer dispersion in pipe flow: testing the theory

Suspensions of microswimmers are a rich source of fascinating new fluid mechanics. Recently we predicted the active pipe flow dispersion of gyrotactic microalgae, whose orientation is biased by gravity and flow shear. Analytical theory predicts that these active swimmers disperse in a markedly distinct manner from passive tracers (Taylor dispersion). Dispersing swimmers display non-zero drift and effective diffusivity that is non-monotonic with Péclet number. Such predictions agree with numerical simulations, but hitherto have not been tested experimentally. Here, to facilitate comparison, we obtain new solutions of the axial dispersion theory accounting both for swimmer negative buoyancy and a local nonlinear response of swimmers to shear, provided by two alternative microscopic stochastic descriptions. We obtain new predictions for suspensions of the model swimming alga Dunaliella salina, whose motility and buoyant mass we parametrise using tracking video microscopy. We then present a new experimental method to measure gyrotactic dispersion using fluorescently stained D. salina and provide a preliminary comparison with predictions of a non-zero drift above the mean flow for each microscopic stochastic description. Finally, we propose further experiments for a full experimental characterisation of gyrotactic dispersion measures and discuss the implications of our results for algal dispersion in industrial photobioreactors.

Design and characterization of a scalable airlift flat panel photobioreactor for microalgae cultivation

A novel flat panel photobioreactor prototype with bulk liquid flow driven by an external airlift was designed, modeled, and experimentally characterized for the purpose of developing scalable industrial photobioreactors. Baffles were built inside the flat panel part of the reactor, directing the liquid bulk flow in a serpentine way, and the external airlift drove the liquid flow and facilitated gas mass transfer. The gas holdup, liquid flow velocity, and oxygen mass transfer of this prototype were experimentally determined and mathematically modeled, and the performance of the reactor was tested by cultivating two species of microalgae, Scenedesmus obliquus and Chlorella sorokiniana. The model-predicted trends correlated well with experimental data, indicating that the reactor might be scaled up using these models. A high cell concentration of C. sorokiniana was achieved under controlled indoor cultivation conditions although serious biofouling occurred in the case of S. obliquus cultivation. The results favor the possibility of scaling up the reactor to industrial scales, based on the models employed, and the potential advantages and disadvantages of the reactor are discussed regarding this industry-oriented photobioreactor configuration in comparison with current industrial photobioreactors.

Event-based predictive control of pH in tubular photobioreactors

This work presents the application of an event-based model predictive control algorithm to regulate the pH in a microalgae production process. The control aim is to maintain the pH within specific limits and to minimize CO2 losses. The control scheme is based on a Generalized Predictive Control (GPC) algorithm with sensor deadband approach. In this algorithm, the controller execution frequency is adapted to the process dynamics. The event-based scheme works with low sampling frequency if controlled variable, pH in this case, is inside an established band. Otherwise, when the pH value is outside the band, the controller actuation frequency is increased to try to drive it quickly near the setpoint based on the selected tolerance. In such a way, the event-based control algorithm allows to establish a tradeoff between control performance and number of process update actions. This fact can be directly related with reduction of CO2 injection times, what is also reflected in CO2 losses. The control structure is first evaluated through simulations using a nonlinear model for microalgal production in tubular photobioreactors. Afterwards, real experiments are presented on an industrial photobioreactor in order to verify the results obtained through simulations. Additionally, the real tests on the industrial plant are used to verify the event-based control scheme in the case of plant-model mismatch, presence of measurement noise and disturbances. Moreover, the control results are compared with classical time-based solutions using well-known control performance indexes.

CO2 mass transfer and conversion to biomass in a horizontal gas–liquid photobioreactor

This study deals with CO2 mass transfers and biomass conversion in an industrial horizontal tubular photobioreactor. An analytical approach is used to determine an expression modeling the influence of CO2 mass transfers on the overall biomass conversion efficiency for a given culture broth, heat and light conditions. Fluid mechanics and mass transfer are predicted with a classical two-phase flow approach (Taitel and Dukler, 1976) combined with a dissolution correlation developed and tested in the laboratory (Valiorgue et al., 2011). The influence of the stripping gas, removing the excess of oxygen in the liquid, on the conversion to biomass efficiency is shown to be not negligible. The expression is used to evaluate how the photobioreactor's design and process parameters can be tuned in order to improve biomass conversion efficiency. The biomass conversion efficiency evolution with the photobioreactor's length was found to behave asymptotically and it was explained by the relative orders of magnitude of gas dissolution and gas stripping. It has been shown that the gas flow rate for stripping and therefore the oxygen removal will be limited when further increasing the industrial photobioreactor's length for a given objective of CO2 conversion to biomass efficiency.

Heterologous stable expression of terpenoid biosynthetic genes using the moss Physcomitrella patens.

Heterologous and stable expression of genes encoding terpenoid biosynthetic enzymes in planta is an important tool for functional characterization and is an attractive alternative to expression in microbial hosts for biotechnological production. Despite improvements to the procedure, such as streamlining of large scale Agrobacterium infiltration and upregulation of the upstream pathways, transient in planta heterologous expression quickly reaches limitations when used for production of terpenoids. Stable integration of transgenes into the nuclear genome of the moss Physcomitrella patens has already been widely recognized as a viable alternative for industrial-scale production of biopharmaceuticals. For expression of terpenoid biosynthetic genes, and reconstruction of heterologous pathways, Physcomitrella has unique attributes that makes it a very promising biotechnological host. These features include a high native tolerance to terpenoids, a simple endogenous terpenoid profile, convenient genome editing using homologous recombination, and cultivation techniques that allow up-scaling from single cells in microtiter plates to industrial photo-bioreactors. Beyond its use for functional characterization of terpenoid biosynthetic genes, engineered Physcomitrella can be a green biotechnological platform for production of terpenoids. Here, we describe two complementary and simple procedures for stable nuclear transformation of Physcomitrella with terpenoid biosynthetic genes, selection and cultivation of transgenic lines, and metabolite analysis of terpenoids produced in transgenic moss lines. We also provide tools for metabolic engineering through genome editing using homologous recombination.