Airlift bioreactor-based strategies for prolonged semi-continuous cultivation of edible Agaricomycetes.

Submerged cultivation of edible filamentous fungi (Agaricomycetes) in bioreactors enables maximum mass transfer of nutrients and has the potential to increase the volumetric productivity of fungal biomass compared to solid state cultivation. These aspects are paramount if one wants to increase the range of bioactives (e.g. glucans) in convenient time frames. In this study, Trametes versicolor (M9911) outperformed four other Agaricomycetes tested strains (during batch cultivations in an airlift bioreactor). This strain was therefore further tested in semi-continuous cultivation. Continuous and semi-continuous cultivations (driven by the dilution rate, D) are the preferred bioprocess strategies for biomass production. We examined the semi-continuous cultivation of T. versicolor at dilution rates between 0.02 and 0.1 h-1. A maximum volumetric productivity of 0.87 g/L/h was obtained with a D of 0.1 h-1 but with a lower total biomass production (cell dry weight, CDW 8.7 g/L) than the one obtained at lower dilution rates (12.3 g/L at D of 0.04 and vs 13.4 g/L, at a D of 0.02 h-1). However, growth at a D of 0.1 h-1 resulted in a very short fermentation (18 h) which terminated due to washout (the specific D exceeded the maximum growth rate of the fungal biomass). At a D of 0.04 h-1, a CDW of 12.3 g/L was achieved without compromising the total residence time (184 h) of the fermentation. While the D of 0.04 h-1 and 0.07 h-1 achieved comparable volumetric productivities (0.5 g/L/h), the total duration of the fermentation at D of 0.07 h-1 was only 85 h. The highest glucan content of cells (27.8 as percentage of CDW) was obtained at a D of 0.07 h-1, while the lowest glucan content was observed in T. versicolor cells grown at a D of 0.02 h-1. KEY POINTS: • The highest reported volumetric productivity for fungal biomass was 0.87 g/L/h. • Semi-continuous fermentation at D of 0.02 h-1 resulted in 13.4 g/L of fungal biomass. • Semi-continuous fermentation at D of 0.07 h-1 resulted in fungal biomass with 28% of total glucans.

A novel physical-biochemical treatment of refinery wastewater.

As of 2022, China has achieved a crude oil processing capacity of 918 million tons, leading to a notable escalation in the production of refinery wastewater. The composition of refinery wastewater is intricate and diverse, posing a substantial challenge to its treatment. In order to facilitate appropriate discharge or reuse, an exhaustive separation process is imperative for refinery wastewater. Conventional pre-treatment processes typically employ inclined plate separators and dissolved air flotation (DAF) for the removal of oil and suspended solids (SS), while sequencing batch reactor (SBR), oxidation ditch, or biological aerated filter (BAF) are employed for the biological treatment process. However, these approaches encounter challenges such as a large spatial footprint, suboptimal treatment efficiency, and high energy consumption. In response to these challenges, this study introduces a novel integrated apparatus consisting of a high-efficiency oil remover (HEOR), coalescence oil remover (COR), and an airlift-enhanced loop bioreactor (AELR). A pilot-scale test was conducted to evaluate the performance of this integrated system in practical field applications. The pilot-scale tests reveal that, without the addition of chemical agents, the petroleum removal efficiency of "HEOR + COR" system was 1.2 times that of DAF. Compared with the SBR system, AELR's volume loading was increased by 1.56 times. The effluent quality achieved in the pilot-scale tests attained parity with that the original process. The "HEOR + COR + AELR" system exhibited energy and carbon emissions reduction of 28% and 30% compared to the "DAF + SBR" system, respectively. Therefore, the operating costs was reduced by approximate 1 Chinese Yuan (CNY) per ton of treated water. This technological advancement serves as a valuable reference for the implementation of low-carbon treatment of refinery wastewater.

Removal of PAHs, TSS, oils and fats from ammonium-rich coke wastewater by granular filtration.

Coke wastewater is a complex industrial wastewater due to its high content of toxic compounds such as cyanides, thiocyanates, phenols, tar, oils, and fats. After a series of treatments, wastewater with a high ammonium content is obtained (around 4,150 mg·L-1). A stripping process is used to reduce it. Certain pollutants in the influent, such as tar, polycyclic aromatic hydrocarbons (PAHs), oils, fats and total suspended solids (TSS), interfere with stripping and therefore must be previously removed. In this study, the performance of a pilot-scale airlift sand filter was evaluated under real conditions for the reduction of the concentration of tar, PAHs, oils, fats and TSS, before stripping. Prior to the sand filter, a cationic flocculant was added to the influent (2 ppm). High (10 mm.min-1), medium (7.5 mm.min-1) and low sand speeds (1.9-2.6 mm.min-1) were assessed. The latter conditions gave the best results: a decrease of 98.2% in TSS, 99.7% in oils, fats and grease and 97.6% in PAHs. The final effluent (≤ 1.6 mg PAHs·L-1, ≤ 5 mg TSS·L-1 and ≤ 0.05 mg·L-1 of fats, oils and grease) was suitable for the stripping process.

Enhancing nitrogen removal from wastewater in a low C/N ratio using an air-lift bio-electrochemical reactor (ALBER).

This study focuses on the development of an air-lift bio-electrochemical reactor (ALBER) with a continuous feeding regime. The objective is to enhance nitrogen removal from synthetic wastewater with a low carbon-to-nitrogen (C/N) ratio. The chemical oxygen demand (COD) and total nitrogen (TN) of the influent wastewater were 500 and 200 mg/L, respectively. The effect of four independent variables, i.e., temperature, hydraulic retention time (HRT), N-NH4+/TN ratio and current density in the range of 16-32 °C, 6-12 h, 25-75%, and 2-10 A/m2, respectively, at three levels on the bio-electrochemical reactor performance were investigated during the bio-electrochemical reactor operation. The Face Center Cube (FCC) of response surface methodology (RSM) was used for design of experiments and model of obtained data. The ALBER achieved the maximum TN removal of 73% (146 mg/l) using external voltage and zeolite/plastic medium at temperature of 16 °C, HRT of 6 h, current density of 2 A/m2 and N-NH4+/TN ratio of 75%. The results indicated that shortening the HRT from 12 to 6 h, reducing the temperature from 32 °C to 24 °C, increasing the current density from 2 to 6 A/m2 and the reduction of nitrate concentration caused an increase in the TN removal. The results indicated that the performance of air-lift bio-electrochemical for nitrogen removal could be attributed to autotrophic denitrification (AD) and simultaneous nitrification/denitrification (SND). The research findings suggest that the ALBER should be further studied for potential use in treating industrial wastewater at low temperatures.

Bioreactor configurations for adventitious root culture: recent advances toward the commercial production of specialized metabolites.

In vitro plant cell and organ cultures are appealing alternatives to traditional methods of producing valuable specialized metabolites for use as: pharmaceuticals, food additives, cosmetics, perfumes, and agricultural chemicals. Cell cultures have been adopted for the production of specialized metabolites in certain plants. However, in certain other systems, adventitious roots are superior to cell suspension cultures as they are organized structures that accumulate high levels of specialized metabolites. The cultivation of adventitious roots has been investigated in various bioreactor systems, including: mechanically agitated, pneumatically agitated, and modified bioreactors. The main relevance and importance of this work are to develop a long-lasting industrial biotechnological technology as well as to improve the synthesis of these metabolites from the plant in vitro systems. These challenges are exacerbated by: the peculiarities of plant cell metabolism, the complexity of specialized metabolite pathways, the proper selection of bioreactor systems, and bioprocess optimization. This review's major objective is to analyze several bioreactor types for the development of adventitious roots, as well as the advantages and disadvantages of each type of bioreactor, and to describe the strategies used to increase the synthesis of specialized metabolites. This review also emphasizes current advancements in the field, and successful instances of scaled-up cultures and the generation of specialized metabolites for commercial purposes are also covered.

Predictive modelling and optimization of an airlift bioreactor for selenite removal from wastewater using artificial neural networks and particle swarm optimization.

Selenite (Se4+) is the most toxic of all the oxyanion forms of selenium. In this study, a feed forward back propagation (BP) based artificial neural network (ANN) model was developed for a fungal pelleted airlift bioreactor (ALR) system treating selenite-laden wastewater. The performance of the bioreactor, i.e., selenite removal efficiency (REselenite) (%) was predicted through two input parameters, namely, the influent selenite concentration (ICselenite) (10 mg/L - 60 mg/L) and hydraulic retention time (HRT) (24 h - 72 h). After training and testing with 96 sets of data points using the Levenberg-Marquardt algorithm, a multi-layer perceptron model (2-10-1) was established. High values of the correlation coefficient (0.96 ≤ R ≤ 0.98), along with low root mean square error (1.72 ≤ RMSE ≤ 2.81) and mean absolute percentage error (1.67 ≤ MAPE ≤ 2.67), clearly demonstrate the accuracy of the ANN model (> 96%) when compared to the experimental data. To ensure an efficient and economically feasible operation of the ALR, the process parameters were optimized using the particle swarm optimization (PSO) algorithm coupled with the neural model. The REselenite was maximized while minimizing the HRT for a preferably higher range of ICselenite. Thus, the most favourable optimum conditions were suggested as: ICselenite - 50.45 mg/L and HRT - 24 h, resulting in REselenite of 69.4%. Overall, it can be inferred that ANN models can successfully substitute knowledge-based models to predict the REselenite in an ALR, and the process parameters can be effectively optimized using PSO.

Oxygen transfer and gas holdup in airlift bioreactors assembled with helical flow promoters.

Bioreactors can perform biochemical conversions mediated by biocatalysts, such as enzymes, animal cells, plants, and microorganisms. Among several existing models, airlift bioreactors are devices with the low shear environment and good mass transfer with low energy consumption, employed in several biochemical processes. The fluid flow is enabled through air injection by the sparger located at the bioreactor base. Despite its simple geometry compared with the conventional bioreactors, airlift performance can be optimized via geometrical modifications. Therefore, the objective of this work was to evaluate the effects of the addition of helical flow promoters, positioned in the riser and/or downcomer regions of an airlift of concentric tubes measuring the volumetric oxygen coefficient (kLa) and gas holdup. The results obtained by varying the gas flow rate from 1.0 to 4.0 vvm allowed the system evaluation of oxygen transfer and gas holdup. The inclusion of helical flow promoters increased the kLa, reaching up to 23% in oxygen transfer compared to tests without helicoids and up to 14% increase in the gas holdup. The inclusion of helical flow promotors was beneficial for all gas flow rates. Thus, including these flow promoters is an effective strategy to increase the oxygen transfer rate for bioprocess optimization.

Production of Recombinant Zika Virus Envelope Protein by Airlift Bioreactor as a New Subunit Vaccine Platform.

The Zika Virus (ZIKV) is an emerging arbovirus of great public health concern, particularly in the Americas after its last outbreak in 2015. There are still major challenges regarding disease control, and there is no ZIKV vaccine currently approved for human use. Among many different vaccine platforms currently under study, the recombinant envelope protein from Zika Virus (rEZIKV) constitutes an alternative option for vaccine development and has great potential for monitoring ZIKV infection and antibody response. This study describes a method to obtain a bioactive and functional rEZIKV using an E. coli expression system, with the aid of a 5-L airlift bioreactor and following an automated fast protein liquid chromatography (FPLC) protocol, capable of obtaining high yields of approximately 20 mg of recombinant protein per liter of bacterium cultures. The purified rEZIKV presented preserved antigenicity and immunogenicity. Our results show that the use of an airlift bioreactor for the production of rEZIKV is ideal for establishing protocols and further research on ZIKV vaccines bioprocess, representing a promising system for the production of a ZIKV envelope recombinant protein-based vaccine candidate.

Obtaining 2,3-Dihydrobenzofuran and 3-Epilupeol from Ageratina pichinchensis (Kunth) R.King & Ho.Rob. Cell Cultures Grown in Shake Flasks under Photoperiod and Darkness, and Its Scale-Up to an Airlift Bioreactor for Enhanced Production.

Ageratina pichinchensis (Kunth) R.King & Ho.Rob. is a plant used in traditional Mexican medicine, and some biotechnological studies have shown that its calluses and cell suspension cultures can produce important anti-inflammatory compounds. In this study, we established a cell culture of A. pichinchensis in a 2 L airlift bioreactor and evaluated the production of the anti-inflammatory compounds 2,3-dihydrobenzofuran (1) and 3-epilupeol (2). The maximum biomass production (11.90 ± 2.48 g/L) was reached at 11 days of culture and cell viability was between 80% and 90%. Among kinetic parameters, the specific growth rate (µ) was 0.2216 days-1 and doubling time (td) was 3.13 days. Gas chromatography coupled with mass spectrometry (GC-MS) analysis of extracts showed the maximum production of compound 1 (903.02 ± 41.06 µg/g extract) and compound 2 (561.63 ± 10.63 µg/g extract) at 7 and 14 days, respectively. This study stands out for the significant production of 2,3-dihydrobenzofuran and 3-epilupeol and by the significant reduction in production time compared to callus and cell suspension cultures, previously reported. To date, these compounds have not been found in the wild plant, i.e., its production has only been reported in cell cultures of A. pichinchensis. Therefore, plant cell cultured in an airlift reactor can be an alternative for the improved production of these anti-inflammatory compounds.

Mitigating membrane fouling in an internal loop airlift membrane photobioreactor containing Spirulina platensis: effects of riser cross-sectional area and hydrophilic baffles.

Membrane photobioreactors (MPBRs) have gained significant attention due to their ability to support microalgae activities such as cultivation, harvesting, and production of beneficial products. Despite various efforts to mitigate membrane fouling, a fundamental issue in membrane processes, in these systems, a cost-effective and less energy-consuming method is still needed. This study examines the impact of the cross-sectional area of the riser and the baffle material on membrane fouling in an internal loop airlift MPBR. The use of hydrophilic polyester-polypropylene (PES-PP) baffles proves to be more effective than plexiglass baffles. Specifically, in configurations with d = 0.7 cm and d = 1.4 cm, RC/RT decreased by approximately 20% and 13%, respectively, compared to plexiglass baffles. As for the values of RP/RT at a distance of d = 0.7, nearly a 5% increase was observed, and at a distance of d = 1.4, an increase of approximately 11% was observed. This is due to the development of the cake layer on the matrix structure of the PES-PP baffles instead of the membrane itself. The most optimal outcomes were reached while working with PES-PP at a distance of 0.7 cm, as it prolonged the membrane fouling time to 46 h.

Effects of temperature, irradiance, and pH on the growth and biochemical composition of Haslea ostrearia batch-cultured in an airlift plan-photobioreactor.

Haslea ostrearia is a pennate diatom that produces marennine, a water-soluble blue pigment responsible for the greening phenomenon and the increase of organoleptic quality of oysters. Apart from the oyster industry, there is a growing interest in the mass cultivation of this diatom due to the biological activities of marennine. To gain knowledge about the feasibility to upscale production of this diatom, in particular, in the context of global warming, the effects of different temperatures (20, 25, and 30 °C), irradiances (100, 200, and 300 µmol photons m-2 s-1), and pH (7.0, 8.0, and 9.0) on growth and biochemical composition were studied in H. ostrearia cultured in an airlift plan-photobioreactor. The maximum growth rate of H. ostrearia (0.9 ± 0.0 day-1) was obtained at 20 °C, 200 µmol photons m-2 s-1, and pH 7.0, referred to as control conditions. The highest concentration in Chla (2.5 ± 0.1 µg 10-6 cells) and total fatty acids (71.6 ± 1.4 mg g-1 of dry weight, DW) was observed at 20 °C, 300 µmol photons m-2 s-1, and pH 7.0. The highest concentration of carotenoids (1.4 ± 0.1 µg 10-6 cells), Chlc (1.3 ± 0.1 µg 10-6 cells), and extracellular marennine (33.1 ± 0.2 µg 10-6 cells) was observed at 30 °C, 200 µmol photons m-2 s-1, and pH 7.0, and a higher protein content (309.7 ± 24.5 mg g-1 of DW) at 25 °C, 200 µmol photons m-2 s-1, and pH 7.0. The biomass of H. ostrearia was enriched with C14:0 fatty acid at 30 °C, 200 µmol photons m-2 s-1, and pH 7.0, and with C16:0 and C16:1n - 7 fatty acids at control conditions. However, DHA C22:6n - 3 (ω-3), C22:0, and C20:0 were only observed at 300 µmol photons m-2 s-1, 20 °C, and pH 7.0. A high abundance of essential polyunsaturated fatty acids C22:1n - 9 (ω-9) + C20:5n - 3 (EPA) was observed at 100 µmol photons m-2 s-1, 20 °C, and pH 7.0. It is thus possible to anticipate and tune the production of specific metabolites through the control of growth conditions of the benthic diatom H. ostrearia. KEY POINTS: • Validation of H. ostrearia culture in a new photobioreactor in batch mode • Biochemical composition variation of H. ostrearia in function of growth conditions • Growth inhibition and unbalanced metabolites induced by the treatment conditions.

Enhanced aerobic granular sludge formation by applying Phanerochaete chrysosporium pellets as induced nucleus.

The long start-up period is a major challenging issue for the widespread application of aerobic granular sludge (AGS). In this study, a novel rapid start-up strategy was developed by inoculating Phanerochaete chrysosporium (P. chrysosporium) pellets as the induced nucleus in a sequencing batch airlift reactor (SBAR) to enhance activated sludge granulation. The results demonstrated that P. chrysosporium pellets could effectively shorten the aerobic granulation time from 32 to 20 days. The AGS promoted by P. chrysosporium pellets had a larger average diameter (2.60-2.74 mm) than that without P. chrysosporium pellets (1.78-1.88 mm) and had better biomass retention capacity and sedimentation properties; its mixed liquor suspended solids (MLSS) and sludge volume index (SVI30) reached approximately 5.2 g/L and 45 mL/g, respectively. The addition of P. chrysosporium pellets promoted the secretion of extracellular polymeric substances (EPS), especially protein (PN). The removal efficiencies of chemical oxygen demand (COD), ammonia nitrogen (NH4+-N), total nitrogen (TN), and total phosphorus (TP) in P. chrysosporium pellets reactor were 98.91%, 89.17%, 64.73%, and 94.42%, respectively, which were higher than those in the reactor without P. chrysosporium pellets (88.73%, 82.09%, 55.75%, and 88.92%). High throughput sequencing analysis indicated that several functional genera that were responsible for the formation of aerobic granules and the removal of pollutants, such as Acinetobacter, Pseudomonas, Janthinobacterium, and Enterobacter, were found to be predominant in the mature sludge granules promoted by P. chrysosporium pellets.

Airlift two-phase partitioning bioreactor for dichloromethane removal: Silicone rubber stimulated biodegradation and its auto-circulation.

Solid non-aqueous phases (NAPs), such as silicone rubber, have been used extensively to improve the removal of volatile organic compounds (VOCs). However, the removal of VOCs is difficult to be further improved because the poor understanding of the mass transfer and reaction processes. Further, the conventional reactors were either complicated or uneconomical. In view of this, herein, an airlift bioreactor with silicone rubber was designed and investigated for dichloromethane (DCM) treatment. The removal efficiency of Reactor 1 (with silicone rubber) was significantly higher than that of Reactor 2 (without silicone rubber), with corresponding higher chloride ion and CO2 production. It was found that Reactor 1 achieved a much better DCM shock tolerance capability and biomass stability than Reactor 2. Silicone rubber not only enhanced the mass transfer in terms of both gas/liquid and gas/microbial phases, but also decreased the toxicity of DCM to microorganisms. Noteworthily, despite the identical inoculum used, the relative abundance of potential DCM-degrading bacteria in Reactor 1 (91.2%) was much higher than that in Reactor 2 (24.3%) at 216 h. Additionally, the silicone rubber could be automatically circulated in the airlift bioreactor due to the driven effect of the airflow, resulting in a significant reduction of energy consumption.

Nutrients recovery from fresh liquid manure through an airlift reactor to mitigate the greenhouse gas emissions of open anaerobic lagoons.

Open anaerobic lagoons are widely used for liquid manure storage and treatment, with excess greenhouse gas (GHG) and odor emissions. In this study, liquid manure was valorized through hybrid nitrogen and phosphorous recovery as value-added products using an airlift reactor. Also, the organic load of liquid manure was reduced before discharging into anaerobic lagoons, which simultaneously mitigated GHG emissions. The results showed that 14.5% of total nitrogen (TN) was recovered as ammonium sulfate, while 38.8% of TN and 79.3% of total phosphorus (TP) were recovered as phosphorus-rich sludge. After the pre-treatment in the reactor, the odor could be controlled effectively due to a 94.2% decrease in total VFAs. In addition, 59.0% of COD was removed, which decreased the theoretical modeled GHG emissions by 51.7% compared to the traditional direct discharging. The application is promising for upgrading anaerobic lagoons of liquid manure.

Lactic acid production by loofah-immobilized Rhizopus oryzae through one-step fermentation process using starch substrate.

Rhizopus oryzae PTCC 5263 capacity in synthesis of lactic acid (LA) from 10 g/l of soluble potato starch was determined using one-step fermentation process. Pellets were the favorable growing form of the free cells. The extent of the natural ability of the test fungus on biofilm formation on loofah sponge was examined by immobilizing R. oryzae (LIRO). The maximum LA concentration for the free cells and LIRO within 96 h was 3 and 4 g/l, respectively. In terms of specific starch utilization rate ([Formula: see text]) and specific LA formation ([Formula: see text]), LIRO performed more favorably compared to the free cells ([Formula: see text] and [Formula: see text]). Cell immobilization strategy was undertaken for the column reactor studies based on the statistically optimized levels of the inoculum size and temperature. Maximum production of the LA by the LIRO using an airlift reactor with net draft tube was 5 g/l obtainable within 48 h.

Multi-factorial analysis of the removal of dichloromethane and toluene in an airlift packing bioreactor.

Biotechnology has proven effective in removing a wide variety of VOCs. In this study, the effects of pH (from 3 to 7), operating temperature (20-30 °C), empty bed residence time (EBRT, 10-40 s) and transient inlet concentration (400-4000 mg m-3) on the removal performance of an airlift packing bioreactor (ALPR) was investigated. The removal efficiency (RE) and stability of the ALPR was evaluated and compared with the conventional airlift bioreactor (ALR). The results showed that under the influence of single factor variation, the ALPR showed significant higher RE and better stability than the ALR in removing dichloromethane (DCM) and toluene. Besides, a factorial design was used to analyses the interaction of multiple factors and their influence on the removal of DCM and toluene in the ALPR and ALR. It shows that pH value has the most significant influence, and plays a crucial role in maintaining high RE of DCM and toluene in both of the ALPR and ALR. Temperature has a great effect on the removal of toluene. EBRT has certain effect on the removal of DCM in the ALPR. The transient concentration of a single substrate has a significant negative effect on the RE of this substrate, while it does not significantly affect the removal of another substrate in the ALPR. However, the steep increase of DCM concentration has an adverse effect on the RE of high concentration toluene in the ALR. The overall RE and degradation capacity of both toluene and DCM by the ALPR are much higher than that of the conventional ALR.

Improving laccase production from Trametes versicolor using lignocellulosic residues as cosubstrates and evaluation of enzymes for blue wastewater biodegradation.

Biocatalytic degradation of recalcitrant pollutants employing ligninolytic enzymes is a promising approach for wastewater treatment. However, enzymes production must be improved to make biodegradation a more cost-effective treatment. In this research, laccase production from Trametes versicolor using lignocellulosic residues (agave bagasse, coconut fibers and wheat bran) as cosubstrates was improved using a central composite face-centered design, and the application of the enzymes-rich culture supernatant was evaluated for blue wastewater biodegradation. Findings revealed that the optimal conditions for laccase production were found at 35 °C and 5 g/L of wheat bran as cosubstrate, reaching about 200 U/mL in 11 days in a batch submerged fermentation. These conditions were scaled up for a submerged fermentation using an airlift reactor, and a maximum enzymatic activity of 1200 U/mL was achieved in 9 days at 30 °C. This enzymes-rich culture supernatant was tested for the degradation of blue wastewater from aircraft in an airlift reactor. Results showed a COD removal efficiency of 43% and an increase of the biodegradability index from 0.64 to 1.36, both results applying an enzymatic activity of supernatant of 300 U/mL. In conclusion, the enzymatic biodegradation becomes a viable strategy for the pretreatment of a real effluent such as the blue wastewater collected in public transportation.

Production of antifungal saponins in an airlift bioreactor with a cell line transformed from Solanum chrysotrichum and its activity against strawberry phytopathogens.

Biotechnology through plant cell cultures in bioreactors is a tool that allows increasing the production of secondary metabolites of commercial interest. The hydrodynamic characterization, in addition to the transfer (OTR) and uptake (OUR) of oxygen through the dynamic method with different aeration rate, were used to see their influence on the production of biomass and saponins. The culture poisoning technique was used to determine the antifungal activity of the SC-2 and SC-3 saponins in vitro. Likewise, the shear or hydrodynamic stress of 273.6 mN/m2 were calculated based on the Reynolds Number. The oxygen supply (OTR) was always greater than the demand (OUR) for all the aeration rate evaluated. Dry weight values of 8.6 gDW/L and a concentration of 2.7 mg/L and 187.3 mg/L of the saponins SC-2 and SC-3 respectively were obtained with an air flow of 0.1 vvm. In addition, it was possible to inhibit the growth of phytopathogenic fungi in vitro by up to 93%, while in vivo it was possible to reduce the infections of strawberry seeds inoculated with phytopathogens, obtaining up to 94% of germinated seeds. This information will facilitate the rational operation of the bioreactor culture system that produces secondary metabolites.

Chlorella vulgaris cultivation in airlift photobioreactor with transparent draft tube: effect of hydrodynamics, light and carbon dioxide on biochemical profile particularly ω-6/ω-3 fatty acid ratio.

Chlorella vulgaris is used for food and feed applications due to its nutraceutical, antioxidant and anticancer properties. An airlift photobioreactor comprising transparent draft tube was used for C. vulgaris cultivation. The effect of reactor parameters like hydrodynamics (0.3-1.5 vvm), light intensity (85-400 µmol m-2 s-1), photoperiod (12-24 h) and gas-phase carbon dioxide (CO2) concentration (5-15% v/v) were evaluated on microalgae and associated bacterial growth, biochemical profile; with special emphasis on ω-3, ω-6 fatty acids, and vitamin B12. The optimal growth of C. vulgaris without CO2 supplementation was observed at 1.2 vvm, which was associated with higher algal productivity, chlorophyll, vitamin B12 content, and bacterial load along with 72% of nitrate removal. The higher light intensity (400 µmol m-2 s-1) and photoperiod (24:0) increased biomass productivity and ω-3 fatty acid content (in lipid) up to 2-3 fold. The elevated levels of gas-phase CO2 concentration (15% v/v) enhanced EPA content up to 7% and biomass productivity up to 171 mg L-1 day-1. However, the increase in CO2 concentration lowered vitamin B12 content (up to 30%) and bacterial load (2-3 log). Also, all the cultivation conditions favoured desirable ω-6/ω-3 ratio(in the range of 1-2).

Outdoor microalgae cultivation in airlift photobioreactor at high irradiance and temperature conditions: effect of batch and fed-batch strategies, photoinhibition, and temperature stress.

The microalgae Scenedesmus abundans cultivated in five identical airlift photobioreactors (PBRs) in batch and fed-batch modes at the outdoor tropical condition. The microalgae strain S. abundans was found to tolerate high temperature (35-45 °C) and high light intensity (770-1690 µmol m- 2 s- 1). The highest biomass productivities were 152.5-162.5 mg L- 1 day- 1 for fed-batch strategy. The biomass productivity was drastically reduced due to photoinhibition effect at a culture temperature of > 45 °C. The lipid compositions showed fatty acids mainly in the form of saturated and monounsaturated fatty acids (> 80%) in all PBRs with Cetane number more than 51. The fed-batch strategies efficiently produced higher biomass and lipid productivities at harsh outdoor conditions. Furthermore, the microalgae also accumulated omega-3 fatty acid (C18:3) up to 14% (w/w) of total fatty acid at given outdoor condition.