Continuous microalgal cultivation in a laboratory-scale photobioreactor under seasonal day–night irradiation: experiments and simulation

In this work, the production of Scenedesmus obliquus in a continuous flat-plate laboratory-scale photobioreactor (PBR) under alternated day–night cycles was tested both experimentally and theoretically. Variation of light intensity according to the four seasons of the year were simulated experimentally by a tunable LED lamp, and effects on microalgal growth and productivity were measured to evaluate the conversion efficiency of light energy into biomass during the different seasons. These results were used to validate a mathematical model for algae growth that can be applied to simulate a large-scale production unit, carried out in a flat-plate PBR of similar geometry. The cellular concentration in the PBR was calculated in both steady-state and transient conditions, and the value of the maintenance kinetic term was correlated to experimental profiles. The relevance of this parameter was finally outlined.     

Theoretical investigation of biomass productivities achievable in solar rectangular photobioreactors for the cyanobacterium Arthrospira platensis

Modeling was done to simulate whole-year running of solar rectangular photobioreactors (PBRs). Introducing the concept of ideal reactor, the maximal biomass productivity that could be achieved on Earth on nitrate as N-source was calculated. Two additional factors were also analyzed with respect to dynamic calculations over the whole year: the effect of PBR location and the effects of given operating conditions on the resulting decrease in productivity compared with the ideal one. Simulations were conducted for the cyanobacterium Arthospira platensis, giving an ideal productivity (upper limit) in the range 55-60 tX ha(-1) year(-1) for a sun tracking system (and around 35-40 tX ha(-1) year(-1) for a fixed horizontal PBR). For an implantation in France (Nantes, west coast), the modification in irradiation conditions resulted in a decrease in biomass productivity of 40%. Various parameters were investigated, with special emphasis on the influence of the incident angle of solar illumination on resulting productivities, affecting both light capture and light transfer inside the bulk culture. It was also found that with appropriate optimization of the residence time as permitted by the model, productivities close to maximal could be achieved for a given location.     

Assessment of key biological and engineering design parameters for production of Chlorella zofingiensis (Chlorophyceae) in outdoor photobioreactors

For the design of a large field of vertical flat plate photobioreactors (PBRs), the effect of four design parameters—initial biomass concentration, optical path length, spacing, and orientation of PBRs—on the biochemical composition and productivity of Chlorella zofingiensis was investigated. A two-stage batch process was assumed in which inoculum is generated under nitrogen-sufficient conditions, followed by accumulation of lipids and carbohydrates in nitrogen-deplete conditions. For nitrogen-deplete conditions, productivity was the most sensitive to initial biomass concentration, as it affects the light availability to individual cells in the culture. An initial areal cell concentration of 50 g m^?2 inoculated into 3.8-cm optical path PBR resulted in the maximum production of lipids (2.42?±?0.02 g m^?2 day^?1) and carbohydrates (3.23?±?0.21 g m^?2 day^?1). Productivity was less sensitive to optical path length. Optical path lengths of 4.8 and 8.4 cm resulted in similar areal productivities (biomass, carbohydrate, and lipid) that were 20 % higher than a 2.4-cm optical path length. Under nitrogen-sufficient conditions, biomass productivity was 48 % higher in PBRs facing north–south during the winter compared to east–west, but orientation had little influence on biomass productivity during the spring and summer despite large differences in insolation. An optimal spacing could not be determined based on growth alone because a tradeoff was observed in which volumetric and PBR productivity increased as space between PBRs increased, but land productivity decreased.     

Kinetic modeling of light limitation and sulfur deprivation effects in the induction of hydrogen production with Chlamydomonas reinhardtii: Part I. Model development and parameter identification

Chlamydomonas reinhardtii is a green microalga capable of turning its metabolism towards H2 production under specific conditions. However this H2 production, narrowly linked to the photosynthetic process, results from complex metabolic reactions highly dependent on the environmental conditions of the cells. A kinetic model has been developed to relate culture evolution from standard photosynthetic growth to H2 producing cells. It represents transition in sulfur-deprived conditions, known to lead to H2 production in Chlamydomonas reinhardtii, and the two main processes then induced which are an over-accumulation of intracellular starch and a progressive reduction of PSII activity for anoxia achievement. Because these phenomena are directly linked to the photosynthetic growth, two kinetic models were associated, the first (one) introducing light dependency (Haldane type model associated to a radiative light transfer model), the second (one) making growth a function of available sulfur amount under extracellular and intracellular forms (Droop formulation). The model parameters identification was realized from experimental data obtained with especially designed experiments and a sensitivity analysis of the model to its parameters was also conducted. Model behavior was finally studied showing interdependency between light transfer conditions, photosynthetic growth, sulfate uptake, photosynthetic activity and O2 release, during transition from oxygenic growth to anoxic H2 production conditions.     

Modeling of Xanthophyllomyces dendrorhous growth on glucose and overflow metabolism in batch and fed-batch cultures for astaxanthin production

An astaxanthin-producing yeast Xanthophyllomyces dendrorhous ENM5 was cultivated in a liquid medium containing 50 g/L glucose as the major carbon source in stirred fermentors (1.5-L working volume) in fully aerobic conditions. Ethanol was produced during the exponential growth phase as a result of overflow metabolism or fermentative catabolism of glucose by yeast cells. After accumulating to a peak of 3.5 g/L, the ethanol was consumed by yeast cells as a carbon source when glucose in the culture was nearly exhausted. High initial glucose concentrations and ethanol accumulation in the culture had inhibitory effects on cell growth. Astaxanthin production was partially associated with cell growth. Based on these culture characteristics, we constructed a modified Monod kinetic model incorporating substrate (glucose) and product (ethanol) inhibition to describe the relationship of cell growth rate with glucose and ethanol concentrations. This kinetic model, coupled with the Luedeking-Piret equation for the astaxanthin production, gave satisfactory prediction of the biomass production, glucose consumption, ethanol formation and consumption, and astaxanthin production in batch cultures over 25-75 g/L glucose concentration ranges. The model was also applied to fed-batch cultures to predict the optimum feeding scheme (feeding glucose and corn steep liquor) for astaxanthin production, leading to a high volumetric yield (28.6 mg/L) and a high productivity (5.36 mg/L/day).     

Optimization of Phaffia rhodozyma continuous culture through response surface methodology

Response surface methodology was applied to optimize the growth of the yeast Phaffia rhodozyma in continuous fermentation using peat hydrolysates as substrate. A second-order, complete, factorial design of the experiments was used to develop empirical models providing a quantitative interpretation of the relationships between the two variables studied, dilution rate and pH. Maximum biomass concentration in the fermentor was obtained by employing the following predicted optimum fermentation conditions: a dilution rate of 0.017/h and a pH level of 7.19. A verification experiment, conducted at previously optimized conditions for maximum biomass volumetric productivity (a dilution rate of 0.022/h, and a pH level of 6.90), produced values for biomass concentration, residual substrate concentration, biomass yield, and biomass volumetric productivity that were very close to the predicted values, indicating the reliability of the empirical model. The concentration of the pigment astaxanthin produced by the yeast under the optimized growth conditions was found to be 544 mg astaxanthin/kg dry cell biomass.     

A theoretical consideration on oxygen production rate in microalgal cultures

Because algal cells are so efficient at absorbing incoming light energy, providing more light energy to photobioreactors would simply decrease energy conversion efficiency. Furthermore, the algal biomass productivity in photobioreactor is always proportional to the total photosynthetic rate. In order to optimize the productivity of algal photobioreactors (PBRs), the oxygen production rate should be estimated. Based on a simple model of light penetration depth and algal photosynthesis, the oxygen production rate in high-density microalgal cultures could be calculated. The estimated values and profiles of oxygen production rate by this model were found to be in accordance with the experimental data. Optimal parameters for PBR operations were also calculated using the model.     

Investigation and modeling of the effects of light spectrum and incident angle on the growth of Chlorella vulgaris in photobioreactors

An in‐depth investigation of how various illumination conditions influence microalgal growth in photobioreactors (PBR) has been presented. Effects of both the light emission spectrum (white and red) and the light incident angle (0° and 60°) on the PBR surface were investigated. The experiments were conducted in two fully controlled lab‐scale PBRs, a torus PBR and a thin flat‐panel PBR for high cell density culture. The results obtained in the torus PBR were used to build the kinetic growth model of Chlorella vulgaris taken as a model species. The PBR model was then applied to the thin flat‐panel PBR, which was run with various illumination conditions. Its detailed representation of local rate of photon absorption under various conditions (spectral calculation of light attenuation, incident angle influence) enabled the model to take into account all the tested conditions with no further adjustment. This allowed a detailed investigation of the coupling between radiation field and photosynthetic growth. Effects of all the radiation conditions together with pigment acclimation, which was found to be relevant, were investigated in depth. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:247–261, 2016     

Effects of acetate on facultative autotrophy in Chlamydomonas reinhardtii assessed by photosynthetic measurements and stable isotope analyses

The green alga Chlamydomonas reinhardtii can grow photoautotrophically utilizing CO(2), heterotrophically utilizing acetate, and mixotrophically utilizing both carbon sources. Growth of cells in increasing concentrations of acetate plus 5% CO(2) in liquid culture progressively reduced photosynthetic CO(2) fixation and net O(2) evolution without effects on respiration, photosystem II efficiency (as measured by chlorophyll fluorescence), or growth. Using the technique of on-line oxygen isotope ratio mass spectrometry, we found that mixotrophic growth in acetate is not associated with activation of the cyanide-insensitive alternative oxidase pathway. The fraction of carbon biomass resulting from photosynthesis, determined by stable carbon isotope ratio mass spectrometry, declined dramatically (about 50%) in cells grown in acetate with saturating light and CO(2). Under these conditions, photosynthetic CO(2) fixation and O(2) evolution were also reduced by about 50%. Some growth conditions (e.g. limiting light, high acetate, solid medium in air) virtually abolished photosynthetic carbon gain. These effects of acetate were exacerbated in mutants with slowed electron transfer through the D1 reaction center protein of photosystem II or impaired chloroplast protein synthesis. Therefore, in mixotrophically grown cells of C. reinhardtii, interpretations of the effects of environmental or genetic manipulations of photosynthesis are likely to be confounded by acetate in the medium.     

Photosynthetic efficiency of Chlamydomonas reinhardtii in attenuated, flashing light

As a result of mixing and light attenuation, algae in a photobioreactor (PBR) alternate between light and dark zones and, therefore, experience variations in photon flux density (PFD). These variations in PFD are called light/dark (L/D) cycles. The objective of this study was to determine how these L/D cycles affect biomass yield on light energy in microalgae cultivation. For our work, we used controlled, short light path, laboratory, turbidostat‐operated PBRs equipped with a LED light source for square‐wave L/D cycles with frequencies from 1 to 100 Hz. Biomass density was adjusted that the PFD leaving the PBR was equal to the compensation point of photosynthesis. Algae were acclimated to a sub‐saturating incident PFD of 220 µmol m^?2 s^?1 for continuous light. Using a duty cycle of 0.5, we observed that L/D cycles of 1 and 10 Hz resulted on average in a 10% lower biomass yield, but L/D cycles of 100 Hz resulted on average in a 35% higher biomass yield than the yield obtained in continuous light. Our results show that interaction of L/D cycle frequency, culture density and incident PFD play a role in overall PBR productivity. Hence, appropriate L/D cycle setting by mixing strategy appears as a possible way to reduce the effect that dark zone exposure impinges on biomass yield in microalgae cultivation. The results may find application in optimization of outdoor PBR design to maximize biomass yields. Biotechnol. Bioeng. 2012; 109: 2567–2574. © 2012 Wiley Periodicals, Inc.     

pH对小球藻Chlorella sp. XQ-200419光合作用、生长和产油的影响

以一种生长快、油脂含量高的小球藻(Chlorella sp. XQ-200419)为实验材料, 利用测定净光合放氧速率的方法研究了pH对其光合作用的影响; 使用改良的BG-11培养基在微藻环形培养池模拟系统中进行分批培养, 培养周期为8d, 培养过程中使用 pH控制仪在线监测藻液的pH, 根据pH变化, 自动接通、关闭CO2通气管道, 将藻液pH分别控制在5.06.0, 7.08.0, 8.09.0, 9.010.0, 10.011.0内, 研究pH对生长速率、生物质面积产率、总脂含量和总脂面积产率的影响。主要结果如下: 藻液pH对小球藻Chlorella sp. XQ-200419光合放氧、生长速率、生物质产率、总脂含量和产率都有显著影响, 适宜的pH范围是7.09.0, 在此范围内, 光合放氧、生长速率、生物质产率、总脂含量和产率均保持较高水平, 且pH的影响不显著; pH低于7.0, 高于9.0, 其光合放氧、生长速率、生物质产率、总脂含量和产率都显著降低。这表明pH对小球藻Chlorella sp. XQ-200419光合作用的影响和对生长、产油的影响是一致的。pH 7.08.0, 小球藻的生物质平均面积产率和总脂平均面积产率都达到最大值, 分别是8.9 g/(m2d)和2269.5 mg/(m2d); 当藻液pH超过10.0, 生物质平均面积产率和总脂平均面积产率分别降低42.1%和60.0%。适合于小球藻生长的pH也有利于其积累油脂, 所以, pH对小球藻产油的影响是一种适宜模式, 而非胁迫模式。规模化培养小球藻Chlorella sp. XQ-200419, 通过补充CO2将藻液pH控制在7.09.0内, 可以获得高生物质产率和总脂产率。研究结果反映出pH对小球藻光合作用、生长和产油影响的规律, 也为规模化培养小球藻生产微藻油脂过程中合理控制藻液pH提供了依据。       

Comparison of two different pneumatically mixed column photobioreactors for the cultivation of Artrospira platensis (Spirulina platensis)

Internal loop airlift and bubble column photobioreactors (PBR) were compared with respect to their performances during cultivation of Artrospira platensis (Spirulina platensis). Culture conditions were kept the same and different parameters were examined through the experiments. It was observed that a higher dry biomass weight and chlorophyll-a concentration was obtained in the airlift PBR yielding a maximum growth rate of 0.45 day(-1), while 0.33 day(-1) was reached in the bubble column PBR. Subsequently, a 17-day of production was carried out in the selected PBR to fully determine the performance of the PBR. Maximum growth rate of 0.47 day(-1) was reached during long term cultivation.     

Optimized aeration by carbon dioxide gas for microalgal production and mass transfer characterization in a vertical flat-plate photobioreactor

To optimize the aeration conditions for microalgal biomass production in a vertical flat-plate photobioreactor (VFPP), the effect of the aeration rate on biomass productivity was investigated under given conditions. Air enriched with 5% or 10% (v/v) CO(2) was supplied for the investigation at rates of 0.025-1 vvm. The CO(2) utilization efficiency, change of pH in the medium, and the optimum aeration rate were determined by evaluating biomass productivity. To investigate the VFPP mass transfer characteristics, the overall volumetric mass transfer coefficient, k(L)a, was evaluated for several different flat-plate sizes. Increasing the height of the VFPP could improve both the mass transfer of CO(2) and the illumination conditions, so this appeared to be a good method for scaling up. Based on a comparison of the k(L)a value at the optimum aeration rate with previously reported results, it was confirmed that the range of CO(2) concentration used in the experiments was cost-effective for mass culture.     

Modeling enzyme production with Aspergillus oryzae in pilot scale vessels with different agitation, aeration, and agitator types

The purpose of this article is to demonstrate how a model can be constructed such that the progress of a submerged fed-batch fermentation of a filamentous fungus can be predicted with acceptable accuracy. The studied process was enzyme production with Aspergillus oryzae in 550 L pilot plant stirred tank reactors. Different conditions of agitation and aeration were employed as well as two different impeller geometries. The limiting factor for the productivity was oxygen supply to the fermentation broth, and the carbon substrate feed flow rate was controlled by the dissolved oxygen tension. In order to predict the available oxygen transfer in the system, the stoichiometry of the reaction equation including maintenance substrate consumption was first determined. Mainly based on the biomass concentration a viscosity prediction model was constructed, because rising viscosity of the fermentation broth due to hyphal growth of the fungus leads to significant lower mass transfer towards the end of the fermentation process. Each compartment of the model was shown to predict the experimental results well. The overall model can be used to predict key process parameters at varying fermentation conditions.     

Biomass productivity of Scenedesmus dimorphus (Chlorophyceae) was improved by using an open pond–photobioreactor hybrid system

The industrialization of microalgae-based biofuel production has been hampered by low biomass productivity of conventional open ponds. In this research, a hybrid cultivation system that combined an open pond and photobioreactor (PBR), with broth circulating between both, was introduced. The hybrid system was tested under indoor and outdoor conditions using the oleaginous microalgal species Scenedesmus dimorphus. When the PBR(s) in the hybrid system reinforced the light supply to the carbon-replete open pond the biomass reached 1.34 g l^–1, 116% higher than in the non-hybrid system. Subsequent studies showed that higher circulation speed and low volume ratio of PBR vs. open pond would further improve the hybrid effects. When applied outdoors at pilot scale, the biomass productivity of the hybrid system increased 46.3–74.3% compared with the open pond and in September was 12.5% higher than that of PBRs. These results indicate that hybrid cultivation might be a cost-effective way to improve the light usage efficiency of current open pond systems.     

Biomass and pigment production for Arthrospira platensis via semi-continuous cultivation in photobioreactors: Temperature effects

This study describes the response of Arthrospira platensis to a variety of temperature conditions as reflected in variations of photosynthetic parameters, pigmentation, and biomass productivity in indoor photobioreactor (PBR) cultivations. These experiments are designed to better understand the impact of temperature, seasonal variations, and acclimation effects on outdoor biomass production. The irradiance level and temperature range (20–39°C) are chosen to enable modeling of semi-continuous operation of large-scale outdoor PBR deployments. Overall, the cultivations are quite stable with some pigment-related instabilities after prolonged high-temperature exposure. Changes in productivity with temperature, as reflected in measured photosynthetic parameters, are immediate and mainly attributable to the temperature dependence of the photosaturation parameter, a secondary factor being variation in pigment content on a longer time scale corresponding to turnover of the culture population. Though pigment changes are not accompanied by significant changes in productivity, prolonged exposure at 35°C and above yields a clear degradation in performance. Productivities in a semi-continuous operation are quantitatively reproduced with a productivity model incorporating photosynthetic parameters measured herein. This study confirms the importance of temperature for biomass and pigment production in Arthrospira cultivations and provides a basis for risk assessments related to temperature mitigation for large-scale outdoor cultivations.     

共生真菌Simplicillium lanosoniveum促进衣藻生长和脂类合成

【背景】藻类是生产生物柴油的主要原料,而一些真菌和细菌能够与藻类共生并提高生物柴油产量,因此藻-菌共生培养技术成为国内外研究的热点。【目的】研究共生真菌Simplicilliumlanosoniveum对衣藻Chlamydomonas reinhardtii细胞生长和脂类合成的影响。【方法】将分离的蓝藻共生真菌和衣藻混合(共生)培养。【结果】与衣藻单独培养相比,混合培养衣藻的比生长速率(0.20 d-1)、细胞产率[0.17 g/(L·d)]和生物量(2.85 g/L)分别提高了10.3%、51.3%和55.7%;脂类比合成速率[0.68 mg/(g·d)]、合成速率[1.95 mg/(L·d)]和含量(220.4 mg/g)分别提高了33.3%、107.5%和32.0%,并且脂类中的饱和脂肪酸以及单不饱和脂肪酸C18-1和C18-2的比例上升,有利于生物柴油的加工。【结论】真菌Simplicilliumlanosoniveum能够促进衣藻的生长和脂类合成,因此藻-菌混合培养可用于生物柴油原料的生产。