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    Nanoliposomal Encapsulation and Purification of AngiotensinConverting Enzyme Inhibitor Peptides from Ulva rigida
    (AMER CHEMICAL SOC, 1155 16TH ST, NW, WASHINGTON, DC 20036, 2025) Şensu, Eda; Koku, Harun; Demircan, Evren; Şişman, Sebahat; Gülseren, İbrahim; Karaduman, Tuğçe; Çakır, Bilal; Okudan, Emine Şükran; Duruksu, Gökhan; Özçelik, Beraat; Yücetepe, Aysun
    Angiotensin-converting enzyme inhibitory peptides derived from natural sources may be effective in the treatment of hypertension without causing side effects compared with existing angiotensin-converting enzyme (ACE) inhibitors. Naturally derived antihypertensive peptides are therefore considered a promising alternative for the prevention or treatment of hypertension. Therefore, the study aimed to purify and identify ACEinhibitory peptides from the green macroalgae Ulva rigida. In addition, the encapsulation of the purified peptides showed the highest ACE-inhibitory activity by chitosan-coated nanoliposomes, and the characterization of nanoliposomes was evaluated. Protein hydrolysates were obtained from U. rigida through enzymatic hydrolysis. The hydrolysates were separated into molecular weights of <3, <5, and <10 kDa through ultrafiltration membrane separation (UFMS). The <3 kDa fraction (UFMS-3) that exhibited the highest ACE-inhibitory activity (77.02%, 1 mg/mL) was purified using ion-exchange chromatography. Fraction-1 (IEC-F1) obtained from the ion-exchange purification showed an impressive 82.03% ACE-inhibitory activity. Moreover, peptide sequences of IEC-F1 were identified by LC-MS/MS, and their bioactive properties were determined in silico. After that, IEC-F1, with a strong ACEinhibitory activity, was loaded into chitosan-coated nanoliposomes to improve their stability for encapsulation. Physical stability (ζpotential, polydispersity index, particle size), thermal (DSC) and morphological properties (SEM), and FT-IR analyses were carried out for the characterization of nanoliposomes. Encapsulation efficiency was found to be 92.0 ± 4.5%. After encapsulation, the ACEinhibitory activity of IEC-F1 was protected by 37.5%. Overall, the obtained findings indicate that the hydrolysate produced by the successive hydrolysis of U. rigida macroalgae with pepsin and trypsin contains peptides with strong ACE-inhibitory action. Furthermore, the chitosan-coated nanoliposome method was determined to be an effective carrier for the delivery of peptide fractions, showing ACE-inhibitory activity. The formulation of chitosan-coated nanoliposomes for peptide fractions from U. rigida represents an innovative approach that allows the development of functional and stable products.
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    Optimization of Protein Extraction from Halopteris scoparia Macroalgae by Ultrasonic-Assisted Enzymatic Extraction (UAEE): Bioactive, Chemical, and Technological Properties
    (Amer Chemical Soc, 2024) Yücetepe, Aysun; Aydar, Elif Feyza; Dogu-Baykut, Esra; Dinç, Hatice; Onat, Isa Alperen; Demircan, Evren; Şensu, Eda
    Finding alternative food sources is now more crucial than ever because of the ever-growing global population, on the one hand, and the changing climate brought on by global warming, on the other. Macroalgae are currently a viable, novel, and alternative food source for the food industry. This research was centered on the optimization of extraction conditions of protein extracts from Halopteris scoparia by the response surface methodology based on the Box-Behnken design. According to the results, extraction yield (18.31-94.5%), total phenolic content (TPC) (11.70-19.07 mg GAE/g dw), and antioxidant activity (AOA(CUPRAC): 11.97-17.85 mg TE/g dw, AOA(ABTS): 45.45-78.02 mg TE/g dw), which were influenced by extraction parameters, were investigated. The optimum extraction conditions were ultrasound application time of 7.46 min, enzyme/substrate of 0.8, and extraction time of 2 h. The water (0.29 +/- 0.07 g/g) and oil absorption capacity (1.37 +/- 0.06 g/g), foaming capacity (8.50 +/- 3.70%) and stability (8.50 +/- 3.70%), and emulsion activity (37.5 +/- 0.00%) and stability (62.22 +/- 7.70%) of the protein extracts obtained under optimum conditions were investigated along with Fourier transform infrared spectroscopy, differential scanning calorimetry, and SDS-PAGE. Protein extracts obtained from H. scoparia showed high TPC and AOA, while the emulsion and foaming properties were found to be lower than those of the algal or plant proteins. The hydrolysis degree of protein hydrolysate from H. scoparia-obtained enzymatic hydrolysis was found to be 61.43%. Sixteen amino acids were identified from the H. scoparia protein hydrolysate. The total amino acid content in the protein hydrolysate was 10.44 mg/g protein. Tyrosine, one of the nonessential amino acids, and leucine and lysine, two of the essential amino acids, were determined as the main amino acids.
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    Orange, red and purple barberries: Effect of in-vitro digestion on antioxidants and ACE inhibitors
    (ELSEVIER, RADARWEG 29, 1043 NX AMSTERDAM, NETHERLANDS, 2021) Şensu, Eda; Kasapoğlu, Kadriye Nur; Gültekin Özgüven, Mine; Demircan, Evren; Arslaner, Ayla; Özçelik, Beraat
    Bioactive compounds such as phenolics, anthocyanins and carotenoids were investigated in Berberis plant with purple (PB), red (RB) and orange (OB) fruits (barberries). The study was aimed to evaluate the bioaccessibility of the antioxidants and ACE (angiotensin converting enzyme) inhibitory compounds of barberry plants upon their transit through in-vitro digestion. Among barberries, (PB) exhibited the strongest antioxidant activity due to its higher phenolic and flavonoid content. The order of total phenolics and anthocyanins was PB > RB > OB. Chlorogenic acid was the main phenolic compound in all barberries even after in-vitro digestion. All barberries had similar amounts of total carotenoids (4.45 mg?-carotene/100gDW). After in-vitro digestion, decreasing phenolics, anthocyanins and carotenoids led to a reduced antioxidant activity. Anthocyanins were found the most sensitive compounds in-vitro digestion. In terms of ACE inhibition activity, only RB exhibited ACE inhibition before (73.84%) and after (65.51%) in-vitro digestion. No ACE inhibition activity was detected at all in RB and OB samples. Leaves and branches of Berberis had also over 50% ACE inhibitory activity. To our knowledge, this is the first study on the carotenoids of barberry and the effect of in-vitro digestion on its bioactive compounds and ACE inhibitors.