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Evaluation of Native Microalgae from Tunisia Using the Pulse-Amplitude-Modulation Measurement of Chlorophyll Fluorescence and A

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Evaluation of Native Microalgae from Tunisia Using the Pulse-Amplitude-Modulation Measurement of Chlorophyll Fluorescence and A View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Qatar University Institutional Repository Jebali et al. Biotechnol Biofuels (2019) 12:119 https://doi.org/10.1186/s13068-019-1461-4 Biotechnology for Biofuels RESEARCH Open Access Evaluation of native microalgae from Tunisia using the pulse-amplitude-modulation measurement of chlorophyll fuorescence and a performance study in semi-continuous mode for biofuel production A. Jebali1,2, F. G. Acién2, N. Jiménez‑Ruiz2, C. Gómez2, J. M. Fernández‑Sevilla2, N. Mhiri1, F. Karray1, S. Sayadi3 and E. Molina‑Grima2* Abstract Background: Microalgae are attracting much attention as a promising feedstock for renewable energy production, while simultaneously providing environmental benefts. So far, comparison studies for microalgae selection for this purpose were mainly based on data obtained from batch cultures, where the lipid content and the growth rate were the main selection parameters. The present study evaluates the performance of native microalgae strains in semi‑ continuous mode, considering the suitability of the algal‑derived fatty acid composition and the saponifable lipid productivity as selection criteria for microalgal fuel production. Evaluation of the photosynthetic performance and the robustness of the selected strain under outdoor conditions was conducted to assess its capability to grow and toler‑ ate harsh environmental growth conditions. Results: In this study, fve native microalgae strains from Tunisia (one freshwater and four marine strains) were isolated and evaluated as potential raw material to produce biofuel. Firstly, molecular identifcation of the strains was performed. Then, experiments in semi‑continuous mode at diferent dilution rates were carried out. The local microal‑ gae strains were characterized in terms of biomass and lipid productivity, in addition to protein content, and fatty acid profle, content and productivity. The marine strain Chlorella sp. showed, at 0.20 1/day dilution rate, lipid and biomass productivities of 35.10 mg/L day and 0.2 g/L day, respectively. Moreover, data from chlorophyll fuorescence measure‑ ments demonstrated the robustness of this strain as it tolerated extreme outdoor conditions including high (38 °C) and low (10 °C) temperature, and high irradiance (1600 µmol/m2 s). Conclusions: Selection of native microalgae allows identifying potential strains suitable for use in the production of biofuels. The selected strain Chlorella sp. demonstrated adequate performance to be scaled up to outdoor conditions. Although experiments were performed at laboratory conditions, the methodology used in this paper allows a robust evaluation of microalgae strains for potential market applications. Keywords: Microalgae, Productivity, Biochemical composition, Biofuel, Chlorophyll fuorescence, Photosynthetic parameters *Correspondence: [email protected] 2 Department of Chemical Engineering, University of Almería, Carretera Sacramento s/n, 04120 Almería, Spain Full list of author information is available at the end of the article © The Author(s) 2019. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creat iveco mmons .org/licen ses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creat iveco mmons .org/ publi cdoma in/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Jebali et al. Biotechnol Biofuels (2019) 12:119 Page 2 of 17 Background which is closer to the industrial commodities produc- Amongst the conventional petroleum substitutes, bio- tion standards and the target must be to maximize both fuel has attracted the greatest interest over the last cen- the biomass and the saponifable lipid productivities tury owing to its sustainability and eco-friendly nature. (i.e., fatty acid methyl esters [FAMEs] productivity) in However, alternative frst- and second-generation bio- addition to the suitability of the algal-derived fatty acid fuel resources, such as crops and lignocellulosic feed- composition [23, 24]. However, few studies have inves- stock, have various drawbacks related to water shortage, tigated these criteria in continuous cultures for select- deforestation, food crop competition and land use [1, 2]. ing the best performing strain and the optimal dilution On the other hand, the judicious exploitation of micro- rate [25–27]. Tis selection approach would allow algae, a third-generation biofuel resource, could over- determining the best combination of the selected crite- come these problems and make a signifcant contribution ria above mentioned. to meeting primary energy demand as well as providing Whatever the operation mode considered, the evalu- environmental benefts [3–6]. In fact, microalgae have a ation of the performance of microalgae cultures is usu- prominent position as a biofuel feedstock given the range ally based on biomass growth; the utilization of fast of advantages they ofer, including a rapid growth rate, no methods such as fuorescence of chlorophylls is usually impact on food security and the ability to grow in poor disregarded. Tis methodology is useful for studying quality waters and on non-arable lands while, at the same photosynthesis mechanisms [28–31]. Te technique time, converting CO2 into carbon-rich lipids [7–9]. Nev- using the Pulse-Amplitude-Modulated fuorometer ertheless, although biofuel production from microalgae is (PAM) is a sensitive tool for examining the variability technically possible, it is currently economically unfeasi- of chlorophyll fuorescence and photosynthetic per- ble due to the high cost of biomass production and pro- formance in microalgae. It allows not only to measure cessing. Additionally, sustainability of biofuel production the photosynthetic activity in a rapid and non-intrusive from freshwater species has been questioned in several mode but also to investigate processes such as photo- reports (http://energ yskep tic.com/2015/algae /). Micro- protection and photoinhibition, the physiological status algae cultivation in seawater is among the few possible of the cell subjected to stress as well as to estimate the options for biomass production for producing fuels at primary production [32–34]. Consequently, the meas- any signifcant scale. Even so, freshwater is still needed to urement of chlorophyll fuorescence allows a rapid and compensate evaporative losses and a consequent increase real-time determination of the photosynthetic perfor- in culture salinity [10]. mance of microalgae cultures under various conditions Extensive studies have been carried out to explore infuencing microalgae growth. For instance, tempera- locally isolated microalgae strains from the northern ture and irradiance are the most important variables for Mediterranean for use in biofuel production [11–15]. controlling microalgae culture growth [35, 36]. Tus, However, exploring the potential of microalgae from these factors are considered among the main environ- the south-western Mediterranean is still in its infancy. mental parameters afecting photosynthesis in outdoor Tunisia is one of the south-western Mediterranean microalgae massive production [37]. countries that possess various biota with untapped In this study, the isolation and molecular identifca- microalgae diversity along with a large non-arable tion of fve native microalgae strains commonly distrib- surface area (totalling almost 40%) as well as suitable uted in north-eastern Tunisia were conducted. To select weather for microalgae growth − 64,000 GJ/ha of solar the best performing strain and the optimal culture radiation a year—making microalgae cultivation for conditions, experiments were carried out in semi-con- biofuel production theoretically feasible [16]. How- tinuous mode. Te biomass productivity, the photosyn- ever, it has been recommended that, prior to large- thetic efciency and the biochemical composition of scale production in a given area, selection of the best the diferent novel isolates were investigated at various performing locally isolated strains should be carried dilution rates, to select the optimal one. In addition, out. Tis is because the native isolates, which are well the robustness and the suitability of the selected strain adapted to local conditions, exhibit better performance for outdoor pilot scale cultivation were evaluated. An and robustness than those from a strain bank collec- assessment of the photosynthetic capacity and parame- tion [17]. Tus, for microalgae strains screening, cur- ters of this microalgae strain towards outdoor environ- rent studies mainly focus on the biomass concentration mental conditions, namely high and low temperature and lipid content, often determined from batch cul- and high light intensity, was carried out by chlorophyll- tures [18–22]. Whereas to adequately select the best fuorescence analysis using the Pulse-Amplitude-Mod- candidate to be used for biofuel production, the study ulated fuorometer (PAM). should be carried out in continuous operational mode, Jebali et al. Biotechnol Biofuels (2019) 12:119 Page 3 of 17 Methods -3′) [40]. Te PCR program for the 18S rRNA gene was Isolation of microalgae strains as previously described [26]. Additionally, the PCR Five diferent strains from north-eastern
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