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Ultrasound and high-pressure assisted extraction of Exopolysaccharides from Phormidium animale: Process optimization via RSM-CCD and evaluation of functional properties

biochemistry
bioengineering
biotechnology

Abstract

This study optimized the extraction of exopolysaccharides (EPS) from the cyanobacterium Phormidium animale using ultrasound-assisted extraction (UAE) and high-pressure-assisted extraction (HPE). Response Surface Methodology (RSM) based on Central Composite Design (CCD) was employed to determine the optimal extraction parameters. For the UAE, the solid-to-liquid ratio, ultrasonic power, and extraction time were varied, while for HPE, pH, pressure, and extraction time were optimized. Both extraction methods were evaluated in terms of EPS yield and structural characteristics. Under optimized conditions, maximum EPS yields of 3.79% (UAE) and 6.26% (HPE) were achieved. The characterization of EPS was performed using nuclear magnetic resonance (NMR) spectroscopy, revealing distinct structural features. The ¹H and ¹³C NMR spectra indicated that UAE primarily extracted simpler compounds with fewer sugar-related signals, while HPE yielded a more complex structure, including additional signals associated with polysaccharides and unsaturated compounds. Scanning electron microscopy (SEM) analysis revealed significant differences in the morphology of EPS, with UAE producing a more porous structure compared to the compact form obtained via HPE. Functional properties, including antioxidant activity, were evaluated through various assays, including DPPH, ABTS, hydroxyl radical scavenging, and ferric ion-reducing power. The EPS demonstrated significant antioxidant activity, particularly in neutralizing DPPH and ABTS radicals, with a concentration-dependent effect observed. The anticancer potential was confirmed through MTT assays, where EPS reduced A549 lung cancer cell viability to 18.07% at 1000 µg/mL, reflecting 93.77% cytotoxicity. The results suggest that UAE and HPE are effective methods for extracting bioactive EPS with promising functional properties, offering potential pharmaceutical applications, nutraceuticals, and biotechnology applications.
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