Today microalgae represent a viable alternative source for high-value products. The specie Chlorella protothecoides (Cp), heterotrophically grown, has been widely studied and provides a high amount of lutein and fatty acids (FA) and has a good profile for biodiesel production. This work studies carotenoid and FA production by autotrophic grown Cp. Cp was grown until the medium's nitrogen was depleted, then diluted in NaCl solution, resulting in nutritional, luminosity, and salinity stresses. Different NaCl concentrations were tested (10, 20, 30 g/L) at two different dilutions. After dilution, a color shifting from green to orange-red was noticed, showing carotenoid production. The best production of both carotenoids and FA was attained with a 20 g/L NaCl solution. The total carotenoid content was 0.8 % w/w (canthaxanthin (23.3 %), echinenone (14.7 %), free astaxanthin (7.1 %), and lutein/zeaxanthin (4.1 %)). Furthermore, the total lipid content reached 43.4 % w/w, with a FA composition of C18:1 (33.64 %), C16:0 (23.30 %), C18:2 (11.53 %), and less than 12 % of C18:3, which is needed to fulfill the biodiesel quality specifications (EN 14214).
This review provides an analysis of recent published work on interactions between microorganisms, especially the ones involving mainly nutrient exchanges and at least with one microalga species. Examples of microbial partners are given, with a remark to the potential application of cultures of an autotroph and a heterotroph, which grow simultaneously, taking advantage of the complementary metabolisms. These are particularly interesting, either due to economic or sustainable aspects, and some applications have already reached the commercial stage of development. The added advantages of these symbiotic cultures are biomass, lipid, and other products productivity enhancement a better utilization of resources and the reduction or even elimination of process residues (including carbon dioxide and other greenhouse gases) to conduct an increasingly greener biotechnology. Among the several symbiotic partners referred, the microalgae and yeast cultures are the most used. The interaction between these two microorganisms shows how to enhance the lipid production for biodiesel purposes compared with separated (stand-alone) cultures.
In order to produce single-cell oil for biodiesel, a yeast and a microalga were, for the first time, grown in two separate reactors connected by their gas-phases, taking advantage of their complementary nutritional metabolisms, i.e., respiration and photosynthesis. The yeast Rhodosporidium toruloides was used for lipid production, originating a carbon dioxide-enriched outlet gas stream which in turn was used to stimulate the autotrophic growth of Chlorella protothecoides in a vertical-alveolar-panel (VAP) photobioreactor. The microalgal biomass productivity was 0.015 gL(-1)h(-1), and its lipid productivity attained 2.2 mgL(-1)h(-1) when aerated with the outlet gas stream from the yeast fermenter. These values represent an increase of 94% and 87%, respectively, as compared to a control culture aerated with air. The CO2 bio-fixed by the microalgal biomass reached an estimated value of 29 mgL(-1)h(-1) in the VAP receiving the gas stream from the fermenter, a value 1.9 times higher than that measured in the control VAP.
^Äëíê~Åí= Multi-parameter flow cytometry was used to monitor cell intrinsic light scatter, viability, and lipid content of `ÜäçêÉää~=éêçíçíÜÉJ ÅçáÇÉë cells grown in shake flasks. Changes in the right angle light scatter (RALS) and forward angle light scatter (FALS) were detected during the microalgal growth, which were attributed to the different microalgal cell cycle stages. The proportion of cells not stained with PI (cells with intact cytoplasmic membrane) was high (> 90%) during the microalgal growth, even in the latter stationary phase, suggesting that the microalgal cells built-up storage materials which allowed them to survive under nutrient starvation, maintaining their cytoplasmic membranes intact. A high correlation between the Nile Red fluorescence intensity measured by flow cytometry and total lipid content assayed by the traditional lipid extraction method was found for this microalga, making this method a suitable and quick technique for the screening of microalgal strains for lipid production, optimization of biofuel production bioprocesses, and scale-up studies. The highest oil content (~28% w/w dry cell weight, estimated by flow cytometry) was observed in the latter stationary phase. In addition, `K=éêçíçíÜÉÅçáÇÉë oil also depicted the adequate fatty acid methyl ester composition for biodiesel purposes at this growth phase, suggesting that the microalgal oil produced during the latter stationary phase could be an adequate substitute for diesel fuel. Medium growth optimization for enhancement of microalgal oil production is now in progress, using the multi-parameter approach. © KSBB hÉóïçêÇëW=Chlorella protothecoidesI=ÖêçïíÜI=äáéáÇëI=ÄáçÑìÉäëI=ãìäíáJé~ê~ãÉíÉê=Ñäçï=ÅóíçãÉíêó
This paper describes the association of two bioreactors: one photoautotrophic and the other heterotrophic, connected by the gas phase and allowing an exchange of O(2) and CO(2) gases between them, benefiting from a symbiotic effect. The association of two bioreactors was proposed with the aim of improving the microalgae oil productivity for biodiesel production. The outlet gas flow from the autotrophic (O(2) enriched) bioreactor was used as the inlet gas flow for the heterotrophic bioreactor. In parallel, the outlet gas flow from another heterotrophic (CO(2) enriched) bioreactor was used as the inlet gas flow for the autotrophic bioreactor. Aside from using the air supplied from the auto- and hetero-trophic bioreactors as controls, one mixotrophic bioreactor was also studied and used as a model, for its claimed advantage of CO(2) and organic carbon being simultaneously assimilated. The microalga Chlorella protothecoides was chosen as a model due to its ability to grow under different nutritional modes (auto, hetero, and mixotrophic), and its ability to attain a high biomass productivity and lipid content, suitable for biodiesel production. The comparison between heterotrophic, autotrophic, and mixotrophic Chlorella protothecoides growth for lipid production revealed that heterotrophic growth achieved the highest biomass productivity and lipid content (>22%), and furthermore showed that these lipids had the most suitable fatty acid profile in order to produce high quality biodiesel. Both associations showed a higher biomass productivity (10-20%), when comparing the two separately operated bioreactors (controls) which occurred on the fourth day. A more remarkable result would have been seen if in actuality the two bioreactors had been inter-connected in a closed loop. The biomass productivity gain would have been 30% and the lipid productivity gain would have been 100%, as seen by comparing the productivities of the symbiotic assemblage with the sum of the two bioreactors operating separately (controls). These results show an advantage of the symbiotic bioreactors association towards a cost-effective microalgal biodiesel production.
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