Towards Sustainable Production of Polybutylene Adipate Terephthalate: Non-biological Catalytic Syntheses of Biomass-derived Constituents.

Renewable chemicals, which are made from renewable resources such as biomass, have attracted significant interest as substitutes for natural gas- or petroleum-derived chemicals to enhance the sustainability of the chemical and petrochemical industries. Polybutylene adipate terephthalate (PBAT), which is a copolyester of 1,4-butanediol (1,4-BDO), adipic acid (AA), and dimethyl terephthalate (DMT) or terephthalic acid (TPA), has garnered significant interest as a biodegradable polymer. This study assesses the non-biological production of PBAT monomers from biomass feedstocks via heterogeneous catalytic reactions. The biomass-based catalytic routes to each monomer are analyzed and compared to conventional routes. Although no fully commercialized catalytic processes for direct conversion of biomass into 1,4-BDO, AA, DMT, and TPA are available, emerging and promising catalytic routes have been proposed. The proposed biomass-based catalytic pathways toward 1,4-BDO, AA, DMT, and TPA are not yet fully competitive with conventional fossil fuel-based pathways mainly due to high feedstock prices and the existence of other alternatives. However, given continuous technological advances in the renewable production of PBAT monomers, bio-based PBAT should be economically viable in the near future.

One-pot, microwave (MW)-assisted production of furfural from almond-, oil-, and wine-derived co-products through biorefinery-based approaches.

This work outlines the first microwave (MW)-assisted protocol for the production of biofuel precursor furfural (FF) from the raw agricultural waste almond hull (AH), olive stone (OS), and the winemaking-derived grape stalk (GS), grape marc (GM) and exhausted grape marc (EGM) through a one-pot synthesis process. To enhance the overall yield, a catalytic process was firstly developed from xylose, major constituent of hemicellulose present in lignocellulosic biomass. This method afforded FF with 100 % selectivity, yielding over 85 % in isolated product when using H2SO4, as opposed to a 37 % yield with AlCl3·6H2O, at 150 °C in only 10 min. For both catalysts, the developed methodology was further validated, proving adaptable and efficient in producing the targeted FF from the aforementioned lignocellulosic raw materials. More specifically, the employment of AlCl3·6H2O resulted in the highest selectivity (up to 89 % from GM) and FF yield (42 % and 39 % molar from OS and AH, respectively), maintaining notable selectivity for the latter (61 and 48 % from AH and OS). At this regard, and considering the environmental factor of sustainability, it is important to point out the role of AlCl3·6H2O in contrast to H2SO4, thus mitigating detrimental substances. This study provides an important management of agricultural waste through sustainable practises for the development of potential bio-based chemicals, aligning with Green Chemistry and process intensification principles.

The green seaweed Ulva: tomorrow's "wheat of the sea" in foods, feeds, nutrition, and biomaterials.

Ulva, a genus of green macroalgae commonly known as sea lettuce, has long been recognized for its nutritional benefits for food and feed. As the demand for sustainable food and feed sources continues to grow, so does the interest in alternative, plant-based protein sources. With its abundance along coastal waters and high protein content, Ulva spp. have emerged as promising candidates. While the use of Ulva in food and feed has its challenges, the utilization of Ulva in other industries, including in biomaterials, biostimulants, and biorefineries, has been growing. This review aims to provide a comprehensive overview of the current status, challenges and opportunities associated with using Ulva in food, feed, and beyond. Drawing on the expertise of leading researchers and industry professionals, it explores the latest knowledge on Ulva's nutritional value, processing methods, and potential benefits for human nutrition, aquaculture feeds, terrestrial feeds, biomaterials, biostimulants and biorefineries. In addition, it examines the economic feasibility of incorporating Ulva into aquafeed. Through its comprehensive and insightful analysis, including a critical review of the challenges and future research needs, this review will be a valuable resource for anyone interested in sustainable aquaculture and Ulva's role in food, feed, biomaterials, biostimulants and beyond.

Harnessing ultrasound-derived hydroxyl radicals for the selective oxidation of aldehyde functions.

Ultrasonic irradiation holds potential for the selective oxidation of non-volatile organic substrates in the aqueous phase by harnessing hydroxyl radicals as chemical initiators. Here, a mechanistic description of hydroxyl radical-initiated glyoxal oxidation is constructed by gleaning insights from photolysis and radiation chemistry to explain the yields and kinetic trends for oxidation products. The mechanistic description and kinetic measurements reported herein reveal that increasing the formation rate of hydroxyl radicals by changing the ultrasound frequency increases both the rates of glyoxal consumption and the selectivity towards C2 acid products over those from C-C cleavage. Glyoxal consumption also occurs more rapidly and with greater selectivity towards C2 acids under acidic conditions, which favor the protonation of carboxylate intermediates into their less reactive acidic forms. Leveraging such pH and frequency effects is crucial to mitigating product degradation by secondary reactions with hydroxyl radicals and oxidation products (specifically hydrogen peroxide and superoxide). These findings demonstrate the potential of ultrasound as a driver for the selective oxidation of aldehyde functions to carboxylic acids, offering a sustainable route for valorizing biomass-derived platform molecules.

Online monitoring lignocellulosic particles by focus beam reflectance measurement for efficient bioprocessing.

Lignocellulose presents a promising alternative to fossil fuels. Monitoring the mass and size changes of lignocellulosic particles without disrupting the process can assist in adjusting pretreatment and enzymatic hydrolysis, where conventional sieving methods fall short. A method utilizing focused beam reflectance measurement (FBRM) was developed to establish mathematical correlations between FBRM chord information (chord length and count) and particle characteristics (weight and size) quantified through sieving. Results indicate particle size exhibits a linear correlation with the square weighted median chord length (Lsqr) with R2 at 0.93. Further, real-time bulk particle mass can be predicted using Lsqr and chord count (R2 0.98). These correlations are applicable in range 53 µm to 358.5 µm. Real-time monitoring of enzymatic hydrolysis of corn stalks has demonstrated the practical applicability of FBRM. This study introduces a novel approach for online characterization of lignocellulosic particles, thereby enhancing lignocellulosic biorefineries.

Simultaneous production of furfural, lignin and cellulose-rich residue from Eucalyptus urophylla × E. grandis by ChCl/1,2-propanediol/MIBK biphasic system pretreatment.

A facile biphasic system composed of choline chloride (ChCl)-based deep eutectic solvent (DES) and methyl isobutyl ketone (MIBK) was developed to realize the furfural production, lignin separation and preparation of fermentable glucose from Eucalyptus in one-pot. Results showed that the ChCl/1,2-propanediol/MIBK system owned the best property to convert hemicelluloses into furfural. Under the optimal conditions (MRChCl:1,2-propanediol = 1:2, raw materials:DES:MIBK ratio = 1:4:8 g/g/mL, 0.075 mol/L AlCl3·6H2O, 140 °C, and 90 min), the furfural yield and glucose yield reached 65.0 and 92.2 %, respectively. Meanwhile, the lignin with low molecular weight (1250-1930 g/mol), low polydispersity (DM = 1.25-1.53) and high purity (only 0.08-2.59 % carbohydrate content) was regenerated from the biphasic system. With the increase of pretreatment temperature, the ß-O-4, ß-ß and ß-5 linkages in the regenerated lignin were gradually broken, and the content of phenolic hydroxyl groups increased, but the content of aliphatic hydroxyl groups decreased. This research provides a new strategy for the comprehensive utilization of lignocellulose in biorefinery process.

A complete review on the oxygen-containing functional groups of biochar: Formation mechanisms, detection methods, engineering, and applications.

Biochar is a porous carbon material generated by the thermal treatment of biomass under anaerobic or anoxic conditions with wealthy Oxygen-containing functional groups (OCFGs). To date, OCFGs of biochar have been extensively studied for their significant utility in pollutant removal, catalysis, capacitive applications, etc. This review adopted a whole system philosophy and systematically summarizes up-to-date knowledge of formation, detection methods, engineering, and application for OCFGs. The formation mechanisms and detection methods of OCFGs, as well as the relationships between OCFGs and pyrolysis conditions (such as feedstocks, temperature, atmosphere, and heating rate), were discussed in detail. The review also summarized strategies and mechanisms for the oxidation of biochar to afford OCFGs, with the performances and mechanisms of OCFGs in the various application fields (environmental remediation, catalytic biorefinery, and electrode material) being highlighted. In the end, the future research direction of biochar OCFGs was put forward.

Simultaneous Optimization and Integration of Multiple Process Heat Cascade and Site Utility Selection for the Design of a New Generation of Sugarcane Biorefinery.

Biorefinery plays a crucial role in the decarbonization of the current economic model, but its high investments and costs make its products less competitive. Identifying the best technological route to maximize operational synergies is crucial for its viability. This study presents a new superstructure model based on mixed integer linear programming to identify an ideal biorefinery configuration. The proposed formulation considers the selection and process scale adjustment, utility selection, and heat integration by heat cascade integration from different processes. The formulation is tested by a study where the impact of new technologies on energy efficiency and the total annualized cost of a sugarcane biorefinery is evaluated. As a result, the energy efficiency of biorefinery increased from 50.25% to 74.5% with methanol production through bagasse gasification, mainly due to its high heat availability that can be transferred to the distillery, which made it possible to shift the bagasse flow from the cogeneration to gasification process. Additionally, the production of DME yields outcomes comparable to methanol production. However, CO2 hydrogenation negatively impacts profitability and energy efficiency due to the significant consumption and electricity cost. Nonetheless, it is advantageous for surface power density as it increases biofuel production without expanding the biomass area.

Metabolic Engineering of Saccharomyces cerevisiae for Production of Canthaxanthin, Zeaxanthin, and Astaxanthin.

The sustainable production of natural compounds is increasingly important in today's industrial landscape. This study investigates the metabolic engineering of Saccharomyces cerevisiae for the efficient biosynthesis of valuable carotenoids: canthaxanthin, zeaxanthin, and astaxanthin. Utilizing a tailored parental yeast strain, Sp_Bc, we optimized the carotenoid pathway by screening and identifying CrtW and CrtZ enzymatic variants. The CrtW variant from Bradyrhizobium sp. achieved a canthaxanthin titer of 425.1 ± 69.1 µg/L, while the CrtZ variant from Pantoea ananatis achieved a zeaxanthin titer of 70.5 ± 10.8 µg/L. Additionally, we optimized carotenoid production by exploring enzyme fusion strategies for all three studied carotenoids and organelle compartmentalization specifically for enhancing astaxanthin synthesis. We further improved carotenoid production by integrating the optimal gene constructs into the yeast genome and deleting the GAL80 gene, enabling the use of sucrose as a carbon source. The engineered strain Sp_Bc-Can001 ∆gal80 was evaluated in a 5 L bioreactor fermentation, achieving a notable canthaxanthin titer of 60.36 ± 1.51 mg/L using sucrose. This research conclusively establishes S. cerevisiae as a viable platform for efficient carotenoid biosynthesis and, for the first time in this yeast system, illustrates sucrose's viability as a carbon source for canthaxanthin production. These findings pave the way for sustainable, cost-effective carotenoid production at an industrial scale.

Innovative Alkanediol-Based Eutectic Solvents for Extracting/Pre-Formulating Dermatologically Valuable Free Fatty Acids from Spirulina and Porphyridium Cakes.

The growing demand for phycobiliproteins from microalgae generates a significant volume of by-products, such as extraction cakes. These cakes are enriched with products of interest for the cosmetics market, namely free fatty acids, particularly polyunsaturated (PUFA). In this work, two cakes, one of spirulina and one of Porphyridium cruentum, were valorized using innovative natural hydrophobic deep eutectic solvents (NaDES) based on alkanediols. The most promising NaDES, as determined by physicochemical properties and screening, are mixtures of alkanediols and fatty acids. These include the mixtures of 1,3-propanediol and octanoic acid (1:5, mol/mol) and 1,3-propanediol and octanoic and decanoic acid (1:3:1, mol/mol). Two extractive processes were implemented: ultrasound-assisted extraction and an innovative mechanical process involving dual asymmetric centrifugation. The second process resulted in the production of extracts significantly enriched in PUFA, ranging from 65 to 220 mg/g dry matter with the two cakes. The extracts and NaDES demonstrated good safety with respect to epidermal keratinocyte viability (>80% at 200 µg/mL). The study of their impact on commensal and pathogenic cutaneous bacteria demonstrated significant effects on the viability of Staphylococcus aureus and Staphylococcus epidermidis (>50% decrease at 200 µg/mL) while preserving Corynebacterium xerosis and Cutibacterium acnes. These results highlight the potential of valorizing these co-products using alkanediol-based NaDES, in a strategy combining an active vector (NaDES) and a growth regulator extract, for the management of cutaneous dysbiosis involving staphylococci.

Potential for curdlan recovery from aerobic granular sludge wastewater treatment systems - A review.

The aerobic granular sludge (AGS) biotechnology has been explored for wastewater treatment for over two decades. AGS is gaining increased interest due to its enhanced treatment performance ability and the potential for resource recovery from AGS-based wastewater treatment systems. Resource recovery from AGS is a promising approach to sustainable wastewater treatment and attaining a circular economy in the wastewater management industry. Currently, research is at an advanced stage on recovering value-added resources such as phosphorus, polyhydroxyalkanoates, alginate-like exopolysaccharides, and tryptophan from waste aerobic granules. Recently, other value-added resources, including curdlan, have been identified in the aerobic granule matrix, and this may increase the sustainability of biotechnology in the wastewater industry. This paper provides an overview of AGS resource recovery potential. In particular, the potential for enhanced curdlan biosynthesis in the granule matrix and its recovery from AGS wastewater treatment systems is outlined.

Integrated Biorefinery for a Next-Generation Methanization Process Focusing on Volatile Fatty Acid Valorization: A Critical Review.

This review addresses the critical issue of a rapidly increasing worldwide waste stream and the need for sustainable management. The paper proposes an integrated transformation toward a next-generation methanization process, which leads not only to treating waste but also to converting it into higher value compounds and greener energy. Although the current and commonly used anaerobic digestion process is useful for biogas production, it presents limitations of resource exploitation and some negative environmental impacts. Focusing on the acidogenic stage in waste stream processing, the paper discusses the recent strategies to enhance the recovery of volatile fatty acids (VFAs). These acids serve as precursors for synthesizing a variety of biochemicals and biofuels, offering higher value products than solely energy recovery and soil fertilizers. Additionally, the importance of recycling the fermentation residues back into the biorefinery process is highlighted. This recycling not only generates additional VFAs but also contributes to generating clean energy, thereby enhancing the overall sustainability and efficiency of the waste management system. Moreover, the review discusses the necessity to integrate life cycle assessment (LCA) and techno-economic analysis (TEA) to evaluate the environmental impacts, sustainability, and processing costs of the proposed biorefinery.

High methane potential of oxygenic photogranules decreases after starvation.

Oxygenic photogranules (OPG) are granular biofilms that can treat wastewater without external aeration, making it an advantage over activated sludge. Excess of OPG biomass can serve as energy source through anaerobic digestion. Two sequencing batch photoreactors were operated over 400 days to grow OPG. Biochemical methane potentials (BMP) were obtained from near-infrared spectroscopy. OPGs had an average BMP of 356 mL CH4·gVS-1, much higher than typical BMP from activated sludge. A partial least squares analysis could relate BMP with reactor operating conditions, like light regime, load or biomass concentration. Since organic load was the most influential parameter on BMP, three starvation experiments were set up. An average decrease of BMP by 18.4 % was observed. However, the unexpected growth of biomass during starvation resulted in a higher total methane volume. In conclusion, starvation reduces the BMP of OPGs but anaerobic digestion of OPG biomass remains a promising route for biomass valorization.

Unveiling the potential of Pseudococcomyxa simplex: a stepwise extraction for cosmetic applications.

Microalgae are gaining attention as they are considered green fabrics able to synthesize many bioactive metabolites, with unique biological activities. However, their use at an industrial scale is still a challenge because of the high costs related to upstream and downstream processes. Here, a biorefinery approach was proposed, starting from the biomass of the green microalga Pseudococcomyxa simplex for the extraction of two classes of molecules with a potential use in the cosmetic industry. Carotenoids were extracted first by an ultrasound-assisted extraction, and then, from the residual biomass, lipids were obtained by a conventional extraction. The chemical characterization of the ethanol extract indicated lutein, a biosynthetic derivative of α-carotene, as the most abundant carotenoid. The extract was found to be fully biocompatible on a cell-based model, active as antioxidant and with an in vitro anti-aging property. In particular, the lutein-enriched fraction was able to activate Nrf2 pathway, which plays a key role also in aging process. Finally, lipids were isolated from the residual biomass and the isolated fatty acids fraction was composed by palmitic and stearic acids. These molecules, fully biocompatible, can find application as emulsifiers and softener agents in cosmetic formulations. Thus, an untapped microalgal species can represent a sustainable source for cosmeceutical formulations. KEY POINTS: • Pseudococcomyxa simplex has been explored in a cascade approach. • Lutein is the main extracted carotenoid and has antioxidant and anti-aging activity. • Fatty acids are mainly composed of palmitic and stearic acids.

Microalgae to bioenergy production: Recent advances, influencing parameters, utilization of wastewater - A critical review.

Fossil fuel limitations and their influence on climate change through atmospheric greenhouse gas emissions have made the excessive use of fossil fuels widely recognized as unsustainable. The high lipid content, carbon-neutral nature and potential as a biofuel source have made microalgae a subject of global study. Microalgae are a promising supply of biomass for third-generation biofuels production since they are renewable. They have the potential to produce significant amounts of biofuel and are considered a sustainable alternative to non-renewable energy sources. Microalgae are currently incapable to synthesize algal biofuel on an extensive basis in a sustainable manner, despite their significance in the global production of biofuels. Wastewater contains nutrients (both organic and inorganic) which is essential for the development of microalgae. Microalgae and wastewater can be combined to remediate waste effectively. Wastewater of various kinds such as industrial, agricultural, domestic, and municipal can be used as a substrate for microalgal growth. This process helps reduce carbon dioxide emissions and makes the production of biofuels more cost-effective. This critical review provides a detailed analysis of the utilization of wastewater as a growth medium for microalgal - biofuel production. The review also highlights potential future strategies to improve the commercial production of biofuels from microalgae.

Microalgae Biomass Production from Rice Husk as Alternative Media Cultivation and Extraction of Phycocyanin Using 3D-Printed Ohmic Heating Reactor.

Phycocyanin is a highly valued pigment present in Spirulina platensis biomass with applications in the food industry in terms of biorefinery concepts; specifically, its antioxidant and antimicrobial capacity are an advantage that could be incorporated into a food matrix. This study aims to use rice husk as an alternative culture medium for S. platensis biomass growth and phycocyanin extraction by ohmic heating processing using a 3D-printed reactor. S. platensis was cultivated in rice husk extract (RHE) from 0-100% (v/v). The highest content of microalgal biomass was 1.75 ± 0.01 g/L, with a specific growth rate of 0.125 ± 0.01 h-1. For the phycocyanin extraction under an ohmic heating process, a 3D-printed reactor was designed and built. To optimize phycocyanin extraction, a central composite rotatable design (CCDR) was evaluated, with three factors: time (min), temperature (°C), and pH. The highest phycocyanin content was 75.80 ± 0.98 mg/g in S. platensis biomass grown with rice husk extract. Ohmic heating is a promising method for rapid phycocyanin extraction, and rice husk as a culture medium is an alternative for the growth of S. platensis biomass in the integration of second- and third-generation biorefineries.

Optimised Degradation of Lignocelluloses by Edible Filamentous Fungi for the Efficient Biorefinery of Sugar Beet Pulp.

The degradation of the complex structure of lignocellulosic biomass is important for its further biorefinery to value-added bioproducts. The use of effective fungal species for the optimised degradation of biomass can promote the effectiveness of the biorefinery of such raw material. In this study, the optimisation of processing parameters (temperature, time, and s/w ratio) for cellulase activity and reducing sugar (RS) production through the hydrolysis of sugar beet pulp (SBP) by edible filamentous fungi of Aspergillus, Fusarium, Botrytis, Penicillium, Rhizopus, and Verticillium spp. was performed. The production of RS was analysed at various solid/water (s/w) ratios (1:10-1:20), different incubation temperatures (20-35 °C), and processing times (60-168 h). The Aspergillus niger CCF 3264 and Penicillium oxalicum CCF 3438 strains showed the most effective carboxymethyl cellulose (CMC) degrading activity and also sugar recovery (15.9-44.8%) from SBP biomass in the one-factor experiments. Mathematical data evaluation indicated that the highest RS concentration (39.15 g/100 g d.w.) and cellulolytic activity (6.67 U/g d.w.) could be achieved using A. niger CCF 3264 for the degradation of SBP at 26 °C temperature with 136 h of processing time and a 1:15 solid/water ratio. This study demonstrates the potential of fungal degradation to be used for SBP biorefining.

One-step ultrasound-assisted recovery of yellow-orange-red natural coloring from defatted annatto seeds: A cleaner processing alternative.

The interest in natural colorants derived from sustainable processes has prompted research into obtaining bixin from defatted annatto (Bixa orellana L.) seeds. Bixin is a compound that imparts yellow-orange-red coloration, known for its high biodegradability, low toxicity, and wide industrial applicability. Meanwhile, high-intensity ultrasound (HIUS) technology has emerged as a promising method for extracting natural colorants, offering higher yields through shorter processes and minimizing thermal degradation. Although some studies have demonstrated the efficiency of HIUS technology in bixin extraction, research on the effects of acoustic cavitation on the properties of the colorant remains limited. Therefore, this study aimed to investigate the influence of HIUS-specific energy levels (0.02, 0.04, 0.12, and 0.20 kJ/g) on the chemical, physical, and morphological characteristics of annatto extracts containing bixin and geranylgeraniol. Single-step extractions of bixin using ethanol as a solvent were evaluated at various acoustic powers (4.6, 8.5, 14.5, and 20 W) and extraction times (0.5, 1, 3, and 5 min) to determine their impact on the yield of natural colorant extraction. Increasing the acoustic power from 4.6 to 20 W and extending the extraction time from 0.5 to 5 min resulted in higher yields of natural colorant, likely due to the effects of acoustic cavitation and increased heat under more intense conditions. However, elevated levels of mechanical and thermal energy did not affect the chemical properties of the colorant, as indicated by UV-Vis and FTIR spectra. Conversely, higher specific energies yielded colorants with a more intense red hue, consistent with increased bixin content, and altered the microstructure and physical state, as observed in X-ray diffractograms. Nevertheless, these alterations did not impact the solubility of the colorant. Therefore, employing a cleaner extraction procedure aided by one-step ultrasound facilitated the recovery of natural colorants and contributed to the biorefining of annatto seeds, enabling the production of a rich geranylgeraniol colorant through a sustainable approach.

Lignocellulolytic Enzymes Production by Four Wild Filamentous Fungi for Olive Stones Valorization: Comparing Three Fermentation Regimens.

Lignocellulolytic enzymes play a crucial role in efficiently converting lignocellulose into valuable platform molecules in various industries. However, they are limited by their production yields, costs, and stability. Consequently, their production by producers adapted to local environments and the choice of low-cost raw materials can address these limitations. Due to the large amounts of olive stones (OS) generated in Morocco which are still undervalued, Penicillium crustosum, Fusarium nygamai, Trichoderma capillare, and Aspergillus calidoustus, are cultivated under different fermentation techniques using this by-product as a local lignocellulosic substrate. Based on a multilevel factorial design, their potential to produce lignocellulolytic enzymes during 15 days of dark incubation was evaluated. The results revealed that P. crustosum expressed a maximum total cellulase activity of 10.9 IU/ml under sequential fermentation (SF) and 3.6 IU/ml of ß-glucosidase activity under submerged fermentation (SmF). F. nygamai recorded the best laccase activity of 9 IU/ml under solid-state fermentation (SSF). Unlike T. capillare, SF was the inducive culture for the former activity with 7.6 IU/ml. A. calidoustus produced, respectively, 1,009 µg/ml of proteins and 11.5 IU/ml of endoglucanase activity as the best results achieved. Optimum cellulase production took place after the 5th day under SF, while ligninases occurred between the 9th and the 11th days under SSF. This study reports for the first time the lignocellulolytic activities of F. nygamai and A. calidoustus. Furthermore, it underlines the potential of the four fungi as biomass decomposers for environmentally-friendly applications, emphasizing the efficiency of OS as an inducing substrate for enzyme production.

Quality of cellulose and biostimulant extracts from Oedogonium calcareum cultivated during primary wastewater treatment.

A practical two-product cascading biorefinery was developed to extract a biostimulant and cellulose from the freshwater filamentous macroalga Oedogonium calcareum grown while treating primary wastewater. Biostimulant production provides a valuable extract with production of disinfected residual biomass for further product development. Both Escherichia coli and F-specific RNA bacteriophage, indicators of human pathogens contamination, were absent from the residual biomass. The chemical composition of the biostimulant was complex, consisting of growth-promoting substances, free amino acids, and minerals. The O. calcareum cellulose fractions yielded between 9.5% and 10.1% (w/w) with purities from 84% to 90% and closely resembled microcrystalline cellulose. Biostimulant extraction improved cellulose quality by increasing crystallinity from 59% to 62%. Biomass condition, drying process, and biostimulant production influenced the crystallinity index. This study demonstrates a two-step process of biostimulant and cellulose extraction from wastewater-grown Oedogonium, simultaneously disinfecting biomass and isolating high-quality cellulose as a sustainable alternative to conventional extraction methods.