Modulation of xanthophyll cycle impacts biomass productivity in the marine microalga Nannochloropsis.

Life on earth depends on photosynthetic primary producers that exploit sunlight to fix CO2 into biomass. Approximately half of global primary production is associated with microalgae living in aquatic environments. Microalgae also represent a promising source of biomass to complement crop cultivation, and they could contribute to the development of a more sustainable bioeconomy. Photosynthetic organisms evolved multiple mechanisms involved in the regulation of photosynthesis to respond to highly variable environmental conditions. While essential to avoid photodamage, regulation of photosynthesis results in dissipation of absorbed light energy, generating a complex trade-off between protection from stress and light-use efficiency. This work investigates the impact of the xanthophyll cycle, the light-induced reversible conversion of violaxanthin into zeaxanthin, on the protection from excess light and on biomass productivity in the marine microalgae of the genus Nannochloropsis. Zeaxanthin is shown to have an essential role in protection from excess light, contributing to the induction of nonphotochemical quenching and scavenging of reactive oxygen species. On the contrary, the overexpression of zeaxanthin epoxidase enables a faster reconversion of zeaxanthin to violaxanthin that is shown to be advantageous for biomass productivity in dense cultures in photobioreactors. These results demonstrate that zeaxanthin accumulation is critical to respond to strong illumination, but it may lead to unnecessary energy losses in light-limiting conditions and accelerating its reconversion to violaxanthin provides an advantage for biomass productivity in microalgae.

Skeletonema marinoi ecotypes show specific habitat-related responses to fluctuating light supporting high potential for growth under photobioreactor light regime.

Diatoms are a diverse group of phytoplankton usually dominating areas characterized by rapidly shifting light conditions. Because of their high growth rates and interesting biochemical profile, their biomass is considered for various commercial applications. This study aimed at identifying strains with superior growth in a photobioreactor (PBR) by screening the natural intraspecific diversity of ecotypes isolated from different habitats. We investigated the effect of PBR light fluctuating on a millisecond scale (FL, simulating the light in a PBR) on 19 ecotypes of the diatom Skeletonema marinoi isolated from the North Sea-Baltic Sea area. We compare growth, pigment ratios, phylogeny, photo-physiological variables and photoacclimation strategies between all strains and perform qPCR and absorption spectra analysis on a subset of strains. Our results show that the ecotypes responded differently to FL, and have contrasting photo-physiological and photoprotective strategies. The strains from Kattegat performed better in FL, and shared common photoacclimation and photoprotection strategies that are the results of adaptation to the specific light climate of the Kattegat area. The strains that performed better with FL conditions had a high light (HL)-acclimated phenotype coupled with unique nonphotochemical quenching features. Based on their characteristics, three strains were identified as good candidates for growth in PBRs.

Moderate high temperature is beneficial or detrimental depending on carbon availability in the green alga Chlamydomonas reinhardtii.

High temperatures impair plant growth and reduce agricultural yields, but the underlying mechanisms remain elusive. The unicellular green alga Chlamydomonas reinhardtii is an excellent model to study heat responses in photosynthetic cells due to its fast growth rate, many similarities in cellular processes to land plants, simple and sequenced genome, and ample genetic and genomics resources. Chlamydomonas grows in light by photosynthesis and with externally supplied acetate as an organic carbon source. Understanding how organic carbon sources affect heat responses is important for the algal industry but remains understudied. We cultivated wild-type Chlamydomonas under highly controlled conditions in photobioreactors at 25 °C (control), 35 °C (moderate high temperature), or 40 °C (acute high temperature) with or without constant acetate supply for 1 or 4 day. Treatment at 35 °C increased algal growth with constant acetate supply but reduced algal growth without sufficient acetate. The overlooked and dynamic effects of 35 °C could be explained by induced acetate uptake and metabolism. Heat treatment at 40 °C for more than 2 day was lethal to algal cultures with or without constant acetate supply. Our findings provide insights to understand algal heat responses and help improve thermotolerance in photosynthetic cells.

Intracellular metabolomic profiling of Picochlorum sp. under diurnal conditions mimicking outdoor light, temperature, and seasonal variations.

INTRODUCTION: This study focuses on metabolic profiling of a robust marine green algal strain Picochlorum sp. MCC39 that exhibits resilient growth under diverse outdoor open pond conditions. Given its potential for producing high-value chemicals through metabolic engineering, understanding its metabolic dynamics is crucial for pathway modification. OBJECTIVES: This study primarily aimed to investigate the metabolic response of Picochlorum sp. to environmental changes. Unlike heterotrophs, algae are subject to diurnal light and temperature, which affect their growth rates and metabolism. Using an environmental photobioreactor (ePBR), we explored how the algal strain adapts to fluctuations in light intensities and temperature within a simulated pond environment. METHODS: We performed a reverse phase ion pairing-LC/MS-MS based metabolome profiling of the MCC39 strain cultivated in simulated pond conditions in ePBR. The experimental setup included diurnal and bi-seasonal variations in light intensities and temperature. RESULTS: The metabolome profile revealed significant differences in 85 metabolites, including amino acids, carboxylic acids, sugar phosphates, purines, pyrimidines, and dipeptides, which exhibited up to 25-fold change in relative concentration with diurnal variations. Seasonal variations also influenced the production of storage molecules, revealing a discernible pattern. The accumulation pattern of metabolites involved in cellular wall formation and energy generation indicated a well-coordinated initiation of photosynthesis and the Calvin cycle with the onset of light. CONCLUSION: The results contribute to a deeper understanding of the adaptability and metabolic response of Picochlorum sp., laying the groundwork for future advancements in algal strain modification.

Determination of nutrient concentration in liquid culture of cyanobacteria Nostoc sp. by capillary electrophoresis and inductively coupled plasma mass spectrometry.

In this work, we demonstrate the use of capillary electrophoresis and inductively coupled plasma mass spectrometry, as competitive methods primarily for ion chromatography, to determine changes in the concentration of small inorganic ions in the Nostoc sp. culture medium. Although macronutrients were analyzed by capillary electrophoresis with conductivity detection, micronutrients were analyzed by inductively coupled plasma mass spectrometry. The different light settings (light intensity and spectral composition) had a visible effect on the culture growth and depletion of calcium, magnesium, and phosphate ions, and iron and manganese elements when comparing the behavior under red or violet light with that under blue light.

Biological performance and membrane fouling of a microalgal-bacterial membrane photobioreactor for wastewater treatment without external aeration and carbonation.

Biological nutrient removal processes involving the use of activated sludge (AS) to treat municipal wastewater normally result in high aeration energy consumption and significant greenhouse gas (GHG) emissions. Therefore, developing cost-efficient and environmentally friendly processes for wastewater treatment is vital. In this work, a novel non-aerated microalgal-bacterial membrane photobioreactor (MB-MPBR) was proposed, and its feasibility for organic contaminant and nutrient removals was evaluated, for the first time. The effects of inoculation ratio (microalgae to bacteria (M/B)) on the biological performance and membrane fouling were systematically investigated. The results showed that 95.9% of the chemical oxygen demand (COD), 74.5% of total nitrogen (TN), 98.5% of NH4+-N and 42.0% of total phosphorus (TP) were removed at an inoculation M/B ratio of 3:2 at steady state, representing a significant improvement compared to the M/B inoculation ratio of 1:3. Additionally, the higher inoculation M/B ratio (3:2) significantly promoted the biomass production owing to the favorable mutual exchange of oxygen and carbon dioxide between microalgae and bacteria. Cake layer formation was the primary fouling mechanism owing to the absence of aeration scouring on the membrane surface. The membrane fouling rate was slightly higher at the higher inoculation ratio (M/B = 3:2) owing to the increased biomass and extracellular polymeric substances (EPS) productions, despite the larger particle size. These results demonstrated that the non-aerated MB-MPBR could achieve superior biological performance, of which the inoculation M/B ratio was of critical importance for the initiation and maintenance of microalgal-bacterial symbiotic system, yet possibly caused severer membrane fouling in the absence of external aeration and carbonation. This study provides a new perspective for further optimizing and applying non-aerated MB-MPBR to enhance municipal wastewater treatment.

Durable photobioreactor antibiofouling coatings for microalgae cultivation by photoreactive poly(2,2,2-trifluoroethyl methacrylate).

To improve the durability of the photobioreactor antibiofouling surface for microalgal cultivation, a series of photoreactive poly(2,2,2-trifluoroethyl methacrylate) (PTFEMA) were successfully synthesized and used to modify ethylene-vinyl acetate (EVA) films by a surface coating and UV light grafting method. Fourier transform infrared (FT-IR) spectra, X-ray photoelectron spectroscopy analysis (XPS) and fluorescence microscopy results indicated that PTFEMA were fixed successfully onto the EVA film surface through a covalent bond. During the microalgal adhesion assay, the number of EVA-PTFEMA film-adhered microalgae was 41.4% lower than that of the EVA film. Moreover, the number of microalgae attached to the EVA-PTFEMA film decreased by 61.7% after cleaning, while that of EVA film decreased by only 49.1%. It was found that the contact angle of EVA-PTFEMA film surface increased, and remained stable when immersed in acid and alkali solution for up to 90 days.HIGHLIGHTSDurable photobioreactor antibiofouling surfaces for microalgal cultivation were prepared successfully.The contact angle of antibiofouling coating surface remained stable in acid and base environment for 90 days.The attached microalgae on antibiofouling surface decreased 41.4% than those of unmodified surface.The attached microalgae on antibiofouling surface could be cleaned by 61.7% through changing the flow velocity of microalgal suspension.

Design parameters comparison of bubble column, airlift and stirred tank photobioreactors for microalgae production.

Microalgae are the most propitious feedstock for biofuel production due to their lipid and fatty acid content. Microalgae cultivation shares many features with bioreactors, such as thermal and pH regulation, feeding procedures, and mixing to enhance heat and mass transfers. Aeration and stirring speeds are important parameters to reduce the costs of producing microalgae. In this study, three different photobioreactor types (stirred tank, airlift, bubble column) were characterized and compared for microalgae production. Hydrodynamics, mass transfer, and power consumption were determined for various aeration rates (0.9, 1.2, 1.5 L/min), and stirring speeds (100, 200 rpm), and Chlorella sorokiniana growth performance was compared under the conditions that provided the highest volumetric mass transfer and the lowest mixing time. Photo-bioreactor homogenization was good as indicated by low mixing times (< 10 s). Bubble column had the highest volumetric mass transfer due to its sparger design. Gas holdup and volumetric mass transfer coefficient were found to increase with the air flow rate and stirring speed. For stirred tank, bubble column, and airlift photobioreactors, maximum specific growth rates of C. sorokiniana were 0.053, 0.061, 0.057 h-1, and biomass productivities were 0.064, 0.097, 0.072 gdw/L.day, respectively. Under the conditions tested, growth was limited by the volumetric mass transfer in the airlift and stirred tank and bubble column was the best option for producing microalgae. These findings pave way for more extensive use of these systems in producing microalgae and provide a basis to compare photobioreactors of different designs.

Biomass productivity and characterization of Tetradesmus obliquus grown in a hybrid photobioreactor.

In this study, the effects of CO2 addition on the growth performance and biochemical composition of the green microalga Tetradesmus obliquus cultured in a hybrid algal production system (HAPS) were investigated. The HAPS combines the characteristics of tubular photobioreactors (towards a better carbon dioxide dissolution coefficient) with thin-layer cascade system (with a higher surface-to-volume ratio). Experimental batches were conducted with and without CO2 addition, and evaluated in terms of productivity and biomass characteristics (elemental composition, protein and lipid contents, pigments and fatty acids profiles). CO2 enrichment positively influenced productivity, and proteins, lipids, pigments and unsaturated fatty acids contents in biomass. The HAPS herein presented contributes to the optimization of microalgae cultures in open systems, since it allows, with a simple adaptation-a transit of the cultivation through a tubular portion where injection and dissolution of CO2 is efficient-to obtain in TLC systems, greater productivity and better-quality biomass.

Microalgae-mediated biofixation as an innovative technology for flue gases towards carbon neutrality: A comprehensive review.

Microalgae-mediated industrial flue gas biofixation has been widely discussed as a clean alternative for greenhouse gas mitigation. Through photosynthetic processes, microalgae can fix carbon dioxide (CO2) and other compounds and can also be exploited to obtain high value-added products in a circular economy. One of the major limitations of this bioprocess is the high concentrations of CO2, sulfur oxides (SOx), and nitrogen oxides (NOx) in flue gases, according to the origin of the fuel, that can inhibit photosynthesis and reduce the process efficiency. To overcome these limitations, researchers have recently developed new technologies and enhanced process configurations, thereby increased productivity and CO2 removal rates. Overall, CO2 biofixation rates from flue gases by microalgae ranged from 72 mg L-1 d -1 to over 435 mg L-1 d-1, which were directly influenced by different factors, mainly the microalgae species and photobioreactor. Additionally, mixotrophic culture have shown potential in improving microalgae productivity. Progress in developing new reactor configurations, with pilot-scale implementations was observed, resulting in an increase in patents related to the subject and in the implementation of companies using combustion gases in microalgae culture. Advancements in microalgae-based green technologies for environmental impact mitigation have led to more efficient biotechnological processes and opened large-scale possibilities.

Microalgae separation in MP-PVC contaminated wastewater using plant-based coagulant over different extraction methods in Bauru, Brazil.

This study investigates the effectiveness of coagulation-flocculation and sedimentation (CFS) for separating microalgae, focusing on the use of various Moringa oleifera extracts as natural coagulants. We examined photobioreactor effluent (PBR) both with and without microplastic PVC (MP-PVC) contamination, referred to as PBR R2 and PBR R1, respectively. Utilising response surface methodology, we identified optimal conditions for the removal of microalgae and MP-PVC. Validation tests demonstrated that the aqueous extract of delipidated Moringa oleifera powder (AEDMOP) achieved high removal efficiencies, with coagulant dosages of 630 mg L-1 for PBR R1 and 625 mg L-1 for PBR R2. Both conditions showed microalgae removal efficiencies exceeding 83% for turbidity, colour, OD540 nm, OD680 nm, and OD750 nm, and 63% for OD254 nm. Interestingly, the optimised conditions for PBR R2 required slightly less coagulant, likely due to the additional particulate matter from MP-PVC. While extracellular polymeric substances (EPS) exhibited a marginal effect on flocculation, further investigation into their role in CFS is necessary. Our findings highlight the potential of AEDMOP for efficient microalgae separation, even in the presence of microplastics.

Unlocking pilot-scale green diesel production from microalgae.

Microalgae have emerged as a promising source of biomass to produce renewable biofuels due to their ability to synthesize high-energy density compounds of commercial interest. This study proposes an approach for pilot-scale oil extraction, purification by fractional distillation, hydrocarbon characterization by gas chromatography coupled to mass spectrometry (GC-MS), evaluation of physicochemical parameters of the produced hydrocarbons, preliminary cost analysis, and challenges and future opportunities for green diesel on a commercial scale. Here, the microalgae Tetradesmus obliquus was cultivated in 12 m³ photobioreactors using biodigested swine waste as a culture medium. The resulting biomass was subjected to drying and harvesting, followed by oil extraction using a hot solvent extraction method, followed by distillation to purify the compounds. Three different extraction and distillation experiments were conducted, each using different solvent combinations. The results obtained revealed that extraction with a solvent blend, composed of hexane and ethanol, provided more significant yields compared to extraction with pure hexane. GC-MS analysis showed the presence of alkanes and alkenes in the oil samples, and the proportion of solvent used in the extraction directly influenced the production of alkanes. Additionally, specific hydrocarbons such as 4-methyl-1-decene, 8-heptadecene, 1-pentadecene, 9-heneicosene, and 2-dodecene were identified. The evaluation of the physicochemical parameters demonstrated that the calorific value of the distilled oil samples is within the range of typical values for petroleum diesel. However, it was observed that the distilled oil samples had higher sulfur content compared to conventional diesel. Regarding the cost analysis, it was found that it varies depending on the experimental conditions. In particular, the process using a solvent mixture of 70% hexane and 30% ethanol proved to be more economical than the others, since it extracted a greater quantity of oil with a lower initial biomass requirement. In summary, this microalgae-derived hydrocarbon production process is promising and offers insights for compound purification and future biofuel applications.

Removal of contaminants of emerging concern from pig manure in different operation stages of a thin-layer cascade photobioreactor. Relationship with concentrations in microalgae and manure.

The performance of a pilot-scale thin-layer cascade photobioreactor, operated in semicontinuous mode, for the removal of veterinary drug residues and other contaminants of emerging concern (CECs) from pig manure has been assessed in six operation stages. Chlorella sp. (70-90%), Scenedesmus sp. (10-25%) and Diatomea (<5%) comprise the microalgae species present during the stages. The global performance to remove the total CEC content in the photobioreactor effluent varied from 62 to 86% on each stage, while an CEC mean amount close to 8% was accumulated in the photobioreactor biomass. A relation with weather conditions was not observed. Elimination ratio was not related to the concentration in the influent which reached up to 8000 ng L-1 for some CECs. As expected, the concentrations of veterinary drugs were higher than those of non-veterinary CECs. The concentrations accumulated in the grown biomass were relative low, lower than 10 ng per fresh g excepting for a few cases. However, statistical data suggested that the linkage of CECs to microalgae biomass boosted their removal from the influent. Furthermore, it was observed that the manure liquid phase contained higher amounts of CECs than the solid phase.

Dialysis bag-microalgae photobioreactor: Novel strategy for enhanced bioresource production and wastewater purification.

Cultivating microalgae in wastewater offers various advantages, but it still faces limitations such as bacteria and other impurities in wastewater affecting the growth and purity of microalgae, difficulty in microalgae harvesting, and extracellular products of microalgae affecting effluent quality. In this study, a novel dialysis bag-microalgae photobioreactor (Db-PBR) was developed to achieve wastewater purification and purer bioresource recovery by culturing microalgae in a dialysis bag. The dialysis bag in the Db-PBR effectively captured the microalgae cells and promoted their lipid accumulation, leading to higher biomass (1.53 times of the control) and lipid production (2.50 times of the control). During the stable operation stage of Db-PBR, the average soluble microbial products (SMP) content outside the dialysis bag was 25.83 mg L-1, which was significantly lower than that inside the dialysis bag (185.63 mg L-1), indicating that the dialysis bag effectively intercepted the SMP secreted by microalgae. As a result, the concentration of dissolved organic carbon (DOC) in Db-PBR effluent was significantly lower than that of traditional photobioreactor. Furthermore, benefiting from the dialysis bag in the reactor effectively intercepted the microorganisms in wastewater, significantly improving the purity of the cultured microalgae biomass, which is beneficial for the development of high-value microalgae products.

A review on the current application of light-emitting diodes for microalgae cultivation and its fiscal analysis.

Microalgae are the promising source of products having a low and high economic value that include feedstock and vitamin supplements. Presently, their cultivation is being carried out by using sunlight in the open raceway ponds. However, this process has disadvantages like fluctuations in irradiance of the sunlight due to climatic changes and bad weather. Artificial lights, exploiting light-emitting diodes are beneficial in increasing the volumetric productivity of the microalgal biomass as it provides continuous illumination in the photobioreactors and assist in the external and internal design. However, the application of light-emitting diodes accrues high input costs. Though the cost of light-emitting diodes was estimated long ago, there is no recent economic analysis of the same. This study aims to enlist the applications of light-emitting diodes in microalgal cultivation with reference to internally illuminated photobioreactors coupled with the evaluation of the cost and energy balance of the artificial lights. The calculation shows that the electrical energy cost incurred during the application of light-emitting diodes for microalgae cultivation is approximately USD 15.19 kg-1 DW. The collective fraction of electrical energy transformed into chemical energy (microalgae biomass) is around 6-8%. The cost of the light-emitting diodes can be decreased by the application of an Arduino-based automated control system to control the power supply to LEDs, photovoltaic powered photobioreactors and additional light. These techniques of input cost reduction have also been explored deeply in the present study. As estimated, they can reduce the cost of light-emitting diodes by 50%.HighlightsDiscussion on the current application of light-emitting diodes for microalgae cultivationA broad discussion on internally illuminated photobioreactors and their modificationsMicroalgae cultivation cost exploiting LEDs' is around USD 15.19 kg-1 DWNet conservation of electrical energy during the cultivation process is 6-8%Photovoltaic powered PBRs and Arduino microcontrollers will decrease cultivation cost.

Bubble column photobioreactor (BCPR) for cultivating microalgae and microalgal consortium (Co-CC) with additional CO2 supply for enhancing biomass, lipid, and preferable fatty acids production.

The present study has designed and developed a 5 L bubble column photobioreactor (BCPR) to investigate two microalgal strains Chlorella sp. S-01, Chlorella sp. S-02 and their consortium Co-CC (Chlorella sp. S-01 + Chlorella sp. S-02) at 0.03, 5, and 10% CO2 supply for biomass and lipid production. The dry cell weight of Chlorella sp. S-01, Chlorella sp. S-02 and Co-CC were, respectively about 1.41, 1.32, and 1.39 g/L on the 20th day without CO2 supply, while it was 1.76, 1.61, and 1.87 g/L, respectively at 10% CO2 supply and similarly, chlorophyll-a content was higher in 10% CO2 supplied condition over control. Interestingly, Co-CC grown at all the CO2 concentrations showed similar lipid content between 19.30 and 1F9.41%. As an integrated refinery approach, de-oiled biomass of Co-CC was subjected to carbohydrates and protein estimation and found that 46.2% and 30.80% in 10% CO2 supply condition in BCPR. Lipid extracted from the Co-CC grown under 0.03, 5, and 10% CO2 supply in 5L BCPR was converted to biodiesel, and the biodiesel yield was estimated to be 62.78%. Further, the fatty acid profile of Co-CC grown at 10% CO2 showed higher levels of C16:0, C16:1, C18:1, and monounsaturated fatty acids contents over other CO2 supplied conditions. Biodiesel of Co-CC showed favourable fuel properties such as density, higher heating value, oxidative stability, CFPP, viscosity, degree of unsaturation, saponification value, and cetane number, which were also in accordance with ASTM, and EN, biodiesel standards.

A biphasic photobioreactor system for consecutive extraction of lipids and carotenoids from pre-hydrolysed microalgae and evaluation of its biodiesel potential.

A green extraction method is developed using partially hydrolysed microalgal cells grown in biocompatible solvent for simultaneous cultivation and extraction of bioproducts from a highly efficient permeabilized microalgal cell with enhanced biomass and lipid content for potential use in biodiesel production. Incomplete digestion of cell wall was achieved by regulating the incubation time of the enzymatic pretreatment of the microalgal cells. 15.77% increase in lipid content was seen when untreated cells were cultured with biocompatible solvent, while cultivation of these enzymatically pretreated cells with biocompatible solvent, the lipid content increased by 53.33% and 22% higher carotenoid content was observed as compared to conventional extraction. The total fatty acids obtained after 1st and 2nd extractions in untreated samples were 67.82%, while those in enzymatically partially digested samples were 91.94%. The untreated and partially enzymatically predigested strain showed suitable properties for quality biodiesel production as per international recommendations. The cost benefit analysis of the overall process showed the use of biocompatible solvent coupled to enzymatically predigested biomass was a favorable option as compared to conventional extraction.

A photobioreactor for production of algae biomass from gaseous emissions of an animal house.

Sustainable approaches to circular economy in animal agriculture are still poorly developed. Here, we report an approach to reduce gaseous emissions of CO2 and NH3 from animal housing while simultaneously using them to produce value-added biomass. To this end, a cone-shaped, helical photobioreactor was developed that can be integrated into animal housing by being freely suspended, thereby combining a small footprint with a physically robust design. The photobioreactor was coupled with the exhaust air of a chicken house to allow continuous cultivation of a mixed culture of Arthrospira spec. (Spirulina). Continuous quantification of CO2 and NH3 concentration showed that the coupled algae reactor effectively purifies the exhaust air from the chicken house while producing algal biomass. Typical production rates of greater than 0.3 g/l*day dry mass were obtained, and continuous operation was possible for several weeks. Morphological, biochemical, and genomic characterization of Spirulina cultures yielded insights into the dynamics and metabolic processes of the microbial community. We anticipate that further optimization of this approach will provide new opportunities for the generation of value-added products from gaseous CO2 and NH3 waste emissions, linking resource-efficient production of microalgae with simultaneous sequestration of animal emissions. KEY POINTS: • Coupling a bioreactor with exhaust gases of chicken coop for production of biomass. • Spirulina mixed culture removes CO2 and NH3 from chicken house emissions. • High growth rates and biodiversity adaptation for nitrogen metabolism. Towards a sustainable circular economy in livestock farming. The functional coupling of a helical tube photobioreactor with exhaust air from a chicken house enabled the efficient cultivation of Spirulina microalgae while simultaneously sequestering the animals' CO2 and NH3 emissions.

Genetic mechanisms underlying increased microalgal thermotolerance, maximal growth rate, and yield on light following adaptive laboratory evolution.

BACKGROUND: Adaptive laboratory evolution (ALE) is a powerful method for strain optimization towards abiotic stress factors and for identifying adaptation mechanisms. In this study, the green microalga Picochlorum sp. BPE23 was cultured under supra-optimal temperature to force genetic adaptation. The robustness and adaptive capacity of Picochlorum strains turned them into an emerging model for evolutionary studies on abiotic stressors such as temperature, salinity, and light. RESULTS: Mutant strains showed an expanded maximal growth temperature of 44.6 °C, whereas the maximal growth temperature of the wild-type strain was 42 °C. Moreover, at the optimal growth temperature of 38 °C, the biomass yield on light was 22.3% higher, and the maximal growth rate was 70.5% higher than the wild type. Genome sequencing and transcriptome analysis were performed to elucidate the mechanisms behind the improved phenotype. A de novo assembled phased reference genome allowed the identification of 21 genic mutations involved in various processes. Moreover, approximately half of the genome contigs were found to be duplicated or even triplicated in all mutants, suggesting a causal role in adaptation. CONCLUSIONS: The developed tools and mutant strains provide a strong framework from whereupon Picochlorum sp. BPE23 can be further developed. Moreover, the extensive strain characterization provides evidence of how microalgae evolve to supra-optimal temperature and to photobioreactor growth conditions. With this study, microalgal evolutionary mechanisms were identified by combining ALE with genome sequencing.

The effect of light emission spectrum on biohydrogen production by Rhodopseudomonas palustris.

Photofermentative hydrogen production has gained increasing attention as a source of green energy. To make such photofermentation processes economically competitive, operating costs need to be reduced, possibly through outdoor operation. Because photofermentation processes are light dependent, the emission spectrum and intensity of light both have a significant influence on the hydrogen production and merit investigation. This study investigates the effect of light sources on the hydrogen production and growth of Rhodopseudomonas palustris, comparing the organism's productivity under longer-wavelength light and light mimicking sunlight. Hydrogen production is enhanced under longer-wavelength light, producing 26.8% (± 7.3%) more hydrogen as compared to under light mimicking that of sunlight; however, R. palustris is still able to produce a considerable volume of hydrogen under light with a spectrum mimicking that of sunlight, providing a promising avenue for future research.