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Air-cushion (AC) packaging has become widely used worldwide. ACs are air-filled, dual plastic packaging solutions commonly found surrounding and protecting items of value within shipping enclosures during transit. Herein, we report on a laboratory assessment employing ACs as a microalgal photobioreactor (PBR). Such a PBR inherently addresses many of the operational issues typically encountered with open raceway ponds and closed photobioreactors, such as evaporative water loss, external contamination, and predation. Using half-filled ACs, the performance of microalgal species Chlorella vulgaris, Nannochloropsis oculata, and Cyclotella cryptica (diatom) was examined and the ash-free dry cell weight and overall biomass productivity determined to be 2.39 g/L and 298.55 mg/L/day for N. oculata, 0.85 g/L and 141.36 mg/L/day for C. vulgaris, and 0.67 g/L and 96.08 mg/L/day for C. cryptica. Furthermore, maximum lipid productivity of 25.54 mg/L/day AFDCW and carbohydrate productivity of 53.69 mg/L/day AFDCW were achieved by C. cryptica, while maximum protein productivity of 247.42 mg/L/day AFDCW was attained by N. oculata. Data from this work will be useful in determining the applicability and life-cycle profile of repurposed and reused ACs as potential microalgal photobioreactors depending upon the end product of interest, scale utilized, and production costs., (© 2023. The Author(s).)

Incorporation of microalgae in biorefineries intended to help society reach carbon neutrality is hindered by algal growth inhibition at high temperatures, necessitating the use of costly and carbon-intensive cooling systems. In the present study, a two-prong strategy of random mutagenesis and adaptive laboratory evolution to generate robust thermotolerant strains of Nannochloropsis oculata, was used. The best mutants demonstrated increased productivity at 35 °C, which was 10 °C higher than the optimal temperature of the wild type. In a 2-L photobioreactor at 35 °C, biomass and lipid productivity were 1.43-fold and 2.24-fold higher, respectively, than wild type at 25 °C. Higher pigment and carbohydrate content contributed to the mutants' rapid growth and enhanced photosynthetic efficiency. Metabolomics and lipidomics showed rewiring of the central carbon metabolism and membrane lipid synthesis in thermotolerant strains to ensure cellular homeostasis without compromising productivity. Tagatose and phosphatidylethanolamine upregulation were identified as future genetic targets for further enhancing lipid production., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

High outdoor temperatures significantly inhibit the growth and lipid production of the industrially promising marine microalga Tetraselmis suecica, which is viewed as a potential feedstock for high-value bioproducts and biofuels. To overcome this limitation, T. suecica was subjected to ultraviolet irradiation to generate mutants capable of being productive at higher temperatures. The top two high-lipid mutants UV-25 and UV-31 isolated at 25 °C and 31 °C, respectively, were compared to the wild type (WT) to delineate physiological alterations and shed light on the mutants' increased biomass and lipid productivity. At 25 °C, UV-25 and UV-31 exhibited lipid productivity of 36.12 and 31.33 mg/L day, which were 1.4- and 1.2-fold higher than WT, respectively. This increase in lipid biosynthesis correlated well with increased carotenoid content in UV-25 (2.2-fold) and UV-31 (3.6-fold), indicating an improved capacity to quench reactive oxygen species. At 31 °C, the growth and lipid accumulation of UV-31 remained high, signifying adaptation to higher temperatures. This is attributed to a well-coordinated modulation of the mutant's cellular metabolism through an increase in galactose and phosphatidylglycerol levels, as well as in protein, all of which contributed to its performance at elevated temperatures. The study successfully established a UV mutagenesis strategy for producing superior- performing microalgae strains with industrially desired traits, paving the way for future outdoor cultivation deployment., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: George P. Philippidis reports financial support was provided by Florida Department of Agriculture and Consumer Services., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Random mutagenesis is an effective strategy for enhancing cellular traits. In this study, we used the mutagen ethyl methanesulfonate to create fast-growing Nannochloropsis oculata mutants. When cultivated in a photobioreactor with a diel cycle, two mutants exhibited 2.2-fold higher carbohydrate productivity and 3.5-4.0-fold higher pigment productivity than the wild type, while one of them also showed 2.5-fold higher lipid productivity. A comprehensive physiological, metabolomic, and lipidomic study showed that the mutants had high levels of glucose-, galactose-, and xylose-based carbohydrates. Their high growth rate was attributed to increased chlorophyll a content, improved nitrogen assimilation, storage, and recycling, and low monogalactosyldiacyl glycerol/digalactosyldiacyl glycerol ratio, which was responsible for higher biomass productivity. The investigation revealed upregulation of lipid precursors, shedding light on high lipid accumulation. The derived algae strains are capable of increasing the biosynthesis of value-added storage molecules without impairing growth, rendering them promising candidates for commercial development in future biorefineries.

The inherent metabolic versatility of Chlorella vulgaris that enables it to metabolize both inorganic and organic carbon under various trophic modes of cultivation makes it a promising candidate for industrial applications. To shed light on the metabolic flexibility of this microalga, time resolved proteomic and metabolomic studies were conducted in three distinct trophic modes (autotrophic, heterotrophic, mixotrophic) at two growth stages (end of linear growth at 6 days and during nutrient deprivation at 10 days). Sweet sorghum bagasse (SSB) hydrolysate was supplied to the cultivation medium as a renewable source of organic carbon mainly in the form of glucose. Integrated multi-omics data showed improved nitrogen assimilation, re-allocation, and recycling and increased levels of photosystem II (PS II) proteins indicating effective cellular quenching of excess electrons during mixotrophy. As external addition of organic carbon (glucose) to the cultivation medium decreases the cell's dependence on photosynthesis, an upregulation in the mitochondrial electron transport chain was recorded that led to increased cellular energy generation and hence higher growth rates under mixotrophy. Moreover, upregulation of the lipid-packaging proteins caleosin and 14_3_3 domain-containing protein resulted in maximum expression during mixotrophy suggesting a strong correlation between lipid synthesis, stabilization, and assembly. Overall, cells cultivated under mixotrophy showed better nutrient stress tolerance and redox balancing leading to higher biomass and lipid production. The study offers a panoramic view of the microalga's metabolic flexibility and contributes to a deeper understanding of the altered biochemical pathways that can be exploited to enhance algal productivity and commercial potential., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The recycling of unfiltered spent media during cultivation of Chlorella vulgaris was studied using metabolomics in an effort to enhance water and nutrient sustainability and reduce operating costs in algal biorefineries. Cultivation in spent media resulted in reduced biomass and lipid productivity by 14% and 19%, respectively, compared to fresh media. The decrease was related to a detected lower nutrient uptake. Nevertheless, carbohydrate content (28% of dry cell weight) and α-linolenic acid content (27 % of fatty acids) were higher in spent media cultures than in fresh media. Metabolomics analysis of intracellular metabolites revealed downregulation of nitrogen assimilation, tricarboxylic acid cycle, structural lipids, and energy metabolism, but upregulation of stress mitigation and carbohydrate synthesis. No growth was supported by spent media during a second cultivation cycle and was likely due to the identified extracellular accumulation of humic acid and free fatty acids that acted as growth auto-inhibitors., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Supplementing cultivation media with exogenous carbon sources enhances biomass and lipid production in microalgae. Utilization of renewable organic carbon from agricultural residues can potentially reduce the cost of algae cultivation, while enhancing sustainability. In the present investigation a medium was developed from sweet sorghum bagasse for cultivation of Chlorella under mixotrophic conditions. Using response surface methodology, the optimal values of critical process parameters were determined, namely inoculum cell density (O.D. 750 ) of 0.786, SSB hydrolysate content of the medium 25% v/v, and zero medium salinity, to achieve maximum lipid productivity of 120 mg/L/d. Enhanced biomass (3.44 g/L) and lipid content (40% of dry cell weight) were observed when the alga was cultivated in SSB hydrolysate under mixotrophic conditions compared to heterotrophic and photoautotrophic conditions. A time course investigation revealed distinct physiological responses in terms of cellular growth and biochemical composition of C. vulgaris cultivated in the various trophic modes. The determined carbohydrate and lipid profiles indicate that sugar addition to the cultivation medium boosts neutral lipid synthesis compared to structural lipids, suggesting that carbon flux is channeled towards triacylglycerol synthesis in the cells. Furthermore, the fatty acid profile of lipids extracted from mixotrophically grown cultures contained more saturated and monosaturated fatty acids, which are suitable for biofuel manufacturing. Scale-up studies in a photobioreactor using SSB hydrolysate achieved a biomass concentration of 2.83 g/L consisting of 34% lipids and 26% carbohydrates. These results confirmed that SSB hydrolysate is a promising feedstock for mixotrophic cultivation of Chlorella and synthesis of algal bioproducts and biofuels.

Landfilling of municipal waste, an environmental challenge worldwide, results in the continuous formation of significant amounts of leachate, which poses a severe contamination threat to ground and surface water resources. Landfill leachate (LL) is generated by rainwater percolating through disposed waste materials and must be treated effectively before safe discharge into the environment. LL contains numerous pollutants and toxic substances, such as dissolved organic matter, inorganic chemicals, heavy metals, and anthropogenic organic compounds. Currently, LL treatment is carried out by a combination of physical, chemical, and microbial technologies. Microalgae are now viewed as a promising sustainable addition to the repertoire of technologies for treating LL. Photosynthetic algae have been shown to grow in LL under laboratory conditions, while some species have also been employed in larger-scale LL treatments. Treating leachate with algae can contribute to sustainable waste management at existing landfills by remediating low-quality water for recycling and reuse and generating large amounts of algal biomass for cost-effective manufacturing of biofuels and bioproducts. In this review, we will examine LL composition, traditional leachate treatment technologies, LL toxicity to algae, and the potential of employing algae at LL treatment facilities. Emphasis is placed on how algae can be integrated with existing technologies for biological treatment of LL, turning leachate from an environmental liability to an asset that can produce value-added biofuels and bioproducts for the bioeconomy.

The present study investigated the effect of light intensity and mixing on microalgae growth in a raceway by comparing the performance of a paddlewheel to a combination of paddlewheel and CO 2 spargers in a 20 L raceway. The increase of light intensity was known to be able to increase the microalgal growth rate. Increasing paddlewheel rotation speed from 13 to 30 rpm enhanced C. vulgaris growth by enhancing culture mixing. Simulation results using computational fluid dynamics (CFD) indicated that both the turnaround areas of the raceway and the area opposite the paddlewheel experienced very low flow velocities (dead zones) of less than 0.1 m/min, which could cause cell settling and slow down growth. The simulated CFD velocity distribution in the raceway was validated by actual velocity measurements. The installation of CO 2 spargers in the dead zones greatly increased flow velocity. The increase of paddlewheel rotation speed reduced the dead zones and hence increased algal biomass production. By complementing the raceway paddlewheel with spargers providing CO 2 at 30 mL/min, we achieved a dry cell weight of 5.2 ± 0.2 g/L, which was about 2.6 times that obtained without CO 2 sparging., (Copyright © 2019 The Society for Biotechnology, Japan. All rights reserved.)

Hydrothermal carbonization converts algal biomass to hydrochar, but also generates a waste aqueous phase rich in nitrogen and other elements that need to be treated before discharge into water bodies. At the same time, cultivation of microalgae requires significant amounts of nutrients, such as nitrogen, phosphorous, and trace metals. Utilization of aqueous phase in algae cultivation can potentially reduce algae production cost, while mitigating waste generation, leading to a circular process that enhances the sustainability of algal biofuels and bioproducts. In the present study, we assessed the potential of aqueous phase generated during hydrothermal carbonization of lipid-extracted algae to support algal biomass and lipid production from Chlorella vulgaris. The aqueous phase was found to contain high concentrations of nitrogen (2.4 g/L), phosphorus (1.75 g/L), organic carbon (20.9 g/L), and trace elements. Among several aqueous phase dilutions in non-sterilized water (2.5–7.5% v/v) tested for algae cultivation, maximum lipid productivity that was 1.7-fold higher than with standard cultivation media was achieved when C. vulgaris was grown in 5% aqueous phase. Time course studies showed significant nutrient removal of 95% nitrate, 60% ammonia, 47% phosphate, and 30% organic carbon by C. vulgaris over the course of a 10-day cultivation. Moreover, when compared with standard media, utilization of the aqueous phase resulted in augmentation of both storage and structural algal lipids that can be used for biofuel and nutraceutical production, respectively. Subsequent bench-top photobioreactor cultivation validated that the aqueous phase is a promising cultivation medium for algal product manufacturing. [Display omitted] • Chlorella vulgaris tolerated up to 7.5% aqueous phase (AP) in water. • Optimal lipid productivity, achieved in 5% AP, exceeded that in standard media. • AP enhanced synthesis of phospholipids and neutral lipids. • AP from hydrothermal carbonization is an effective nutrient source for algae. [ABSTRACT FROM AUTHOR]

Microalgae are increasingly viewed as renewable biological resources for a wide range of chemical compounds that can be used as or transformed into biomaterials through biorefining to foster the bioeconomy of the future. Besides the well-established biofuel potential of microalgae, key microalgal bioactive compounds, such as lipids, proteins, polysaccharides, pigments, vitamins, and polyphenols, possess a wide range of biomedical and nutritional attributes. Hence, microalgae can find value-added applications in the nutraceutical, pharmaceutical, cosmetics, personal care, animal food, and agricultural industries. Microalgal biomass can be processed into biomaterials for use in dyes, paints, bioplastics, biopolymers, and nanoparticles, or as hydrochar and biochar in solid fuel cells and soil amendments. Equally important is the use of microalgae in environmental applications, where they can serve in heavy metal bioremediation, wastewater treatment, and carbon sequestration thanks to their nutrient uptake and adsorptive properties. The present article provides a comprehensive review of microalgae specifically focused on biomaterial production and environmental applications in an effort to assess their current status and spur further deployment into the commercial arena. [ABSTRACT FROM AUTHOR]

The growing demand of microalgal biomass for biofuels, nutraceuticals, cosmetics, animal feed, and other bioproducts has created a strong interest in developing low-cost sustainable cultivation media and methods. Agricultural and food residues represent low-cost abundant and renewable sources of organic carbon that can be valorized for the cultivation of microalgae, while converting them from an environmental liability to an industrial asset. Biochemical treatment of such residues results in the release of various sugars, primarily glucose, sucrose, fructose, arabinose, and xylose along with other nutrients, such as trace elements. These sugars and nutrients can be metabolized in the absence of light (heterotrophic) or the presence of light (mixotrophic) by a variety of microalgae species for biomass and bioproduct production. The present review provides an up-to-date critical assessment of the prospects of various types of agricultural and food residues to serve as algae feedstocks and the microalgae species that can be grown on such residues under a range of cultivation conditions. Utilization of these feedstocks can create potential industrial applications for sustainable production of microalgal biomass and bioproducts. [ABSTRACT FROM AUTHOR]

Oleaginous microalgae and yeasts represent promising candidates for large-scale production of lipids, which can be utilized for production of drop-in biofuels, nutraceuticals, pigments, and cosmetics. However, low lipid productivity and costly downstream processing continue to hamper the commercial deployment of oleaginous microorganisms. Strain improvement can play an essential role in the development of such industrial microorganisms by increasing lipid production and hence reducing production costs. The main means of strain improvement are random mutagenesis, adaptive laboratory evolution (ALE), and rational genetic engineering. Among these, random mutagenesis and ALE are straight forward, low-cost, and do not require thorough knowledge of the microorganism's genetic composition. This paper reviews available mutagenesis and ALE techniques and screening methods to effectively select for oleaginous microalgae and yeasts with enhanced lipid yield and understand the alterations caused to metabolic pathways, which could subsequently serve as the basis for further targeted genetic engineering. [ABSTRACT FROM AUTHOR]

Microalgae are increasingly viewed as a sustainable potential resource of chemicals and biologicals. The production of light alkenes (C2-C4 olefins), a major precursor to plastics and other value-added chemicals currently produced from fossil fuels, was studied by catalytically cracking marine microalgae Picochlorum oculatum . The process was studied in the temperature range of 400–650 °C using two aluminosilicate catalysts; (1) commercial Si/Al at catalyst-to-algae mass ratios of zero, 1:1, 5:1, 10:1, and 20:1; and (2) H-β zeolite at a catalyst-to-algae mass ratio of 10:1. Ethene was obtained as the most significant light olefin product followed by propene and butenes as a result of the selectivity of the Si/Al catalyst towards ethene formation. The total olefin yield rose with increasing temperature, as cracking of cellular macromolecules intensified. However, olefin formation started to level off above 550 °C. The total olefin yield also rose with increasing catalyst-to-algae mass ratio, as a result of the presence of more catalytic reaction sites, but the impact lessened at high ratios. A maximum ethene yield (7.3%) was achieved using Si/Al at a catalyst-to-algae mass ratio of 20:1, whereas a maximum propene yield (5.1%) was obtained using H-β zeolite at a catalyst-to-algae ratio of 10:1. The study provided a more effective catalyst for producing light olefins, compared to previous reports, as the maximum total olefin yields of 10.3% and 12.3% with Si/Al (20:1 ratio) and H-β zeolite (10:1 ratio), respectively, exceeded the literature reported yield by 34% and 60%, respectively. [ABSTRACT FROM AUTHOR]