| Summary: | Enzyme-assisted extraction (EAE) and ultrasound-assisted extraction (UAE) are both recognized as sustainable processes, but little has been done on the combined process known as ultrasound-assisted enzymatic hydrolysis (UAEH), and even less on seaweed. The present study aimed to optimize the UAEH of the red seaweed <i>Grateloupia turuturu</i> for the extraction of R-phycoerythrin (R-PE) directly from the wet biomass by applying a response surface methodology based on a central composite design. Three parameters were studied: the power of ultrasound, the temperature and the flow rate in the experimental system. Data analysis demonstrated that only the temperature had a significant and negative effect on the R-PE extraction yield. Under the optimized conditions, the R-PE kinetic yield reached a plateau between 90 and 210 min, with a yield of 4.28 ± 0.09 mg·g<sup>−1</sup> dry weight (dw) at 180 min, corresponding to a yield 2.3 times higher than with the conventional phosphate buffer extraction on freeze-dried <i>G. turuturu</i>. Furthermore, the increased release of R-PE, carbohydrates, carbon and nitrogen can be associated with the degradation of <i>G. turuturu</i> constitutive polysaccharides, as their average molecular weights had been divided by 2.2 in 210 min. Our results thus demonstrated that an optimized UAEH is an efficient method to extract R-PE from wet <i>G. turuturu</i> without the need for expensive pre-treatment steps found in the conventional extraction. UAEH represents a promising and sustainable approach that should be investigated on biomasses where the recovery of added-value compounds needs to be improved.
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