Multifunctional natural derived peptide extracts for skin care

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In vitro gastrointestinal digestion impact on the antioxidant activity of extracts produced from the microalgae chlorella vulgaris and nannochloropsis oceanica

Publication . Cunha, Sara A.; Salsinha, Ana S.; Silva, Joana Laranjeira; Pintado, Manuela

Microalgae are rapidly cultivable growing photosynthetic organisms rich in compounds with high biological values, such as antioxidants, with interest for food industry. Although microalgae extracts may present several health benefits, it is necessary to understand if their properties are maintained throughout the gastrointestinal (GI) digestion, after the exposure to enzymes and different pH ranges. Thus, this work studied the impact of in vitro gastrointestinal digestion on the antioxidant activity of extracts produced from the microalgae Chlorella vulgaris and Nannochloropsis oceanica, with the goal of evaluating their potential as functional food ingredients. Chlorella extract was produced by acid and enzymatic hydrolysis (cellulase and subtilisin) and Nannochloropsis extract by enzymatic hydrolysis (cellulase and subtilisin). Both were submitted to simulated GI conditions. The antioxidant activity was determined by ORAC and ABTS assays in four stages of GI simulation (before digestion, and after mouth, stomach and intestine digestion). Both extracts showed increased ORAC and ABTS values throughout the GI digestion. This increase was statistically significant for Chlorella in terms of ABTS values in all phases, and in ORAC after stomach and intestine digestion. For Nannochloropsis, only the increase in ABTS values was statistically different after stomach and intestine digestion (pThis study showed that both extracts maintain their antioxidant activity throughout in vitro GI digestion, with a little increase being observed, which may be explained by the formation of smaller and more bioactive peptides. These results shows that Chlorella and Nannochloropsis extracts may be considered sustainable antioxidant ingredients for the development of functional food.

2021-07Journal article

Open access

Bioactive peptides produced from the mussel mytilus galloprovincialis by enzymatic hydrolysis with corolase

Publication . Cunha, S. A.; Castro, R. de; Coscueta, E. R.; Pintado, M. E.

Bioactive peptides have been produced from several marine sources since they can exhibit positive effects for humans and animals. Mytilus galloprovincialis is highly consumed in several countries and has a meat rich in proteins, which has been described as a source of bioactive peptides with relevant properties such as antioxidant, anti-hypertensive and antimicrobial activities. Mussel commercialization generates waste, since the small or broken mussels are discarded. Thus, in this work discarded mussels were used with the goal of producing water soluble extracts rich in proteins and bioactive peptides. The mussel meat was firstly minced, and then submitted to different conditions of enzymatic hydrolysis, using the protease corolase to generate peptides. The variable factors evaluated were temperature, incubation time and enzyme concentration. To achieve the optimal extraction conditions, a Box-Behnken experimental design was performed using statgraphic centurion software. Factorial design allowed the evaluation of the effects of the three factors on protein release, antioxidant and anti-hypertensive properties of the extracts. The protein content of each extract was determined by Kjeldahl, the antioxidant activity was determined by oxygen-radical absorbance-capacity (ORAC) assay and anti-hypertensive property was determined by the inhibition method of Angiotensin-I converting enzyme (ACE). The incubation of the minced mussel meat with 3% of enzyme, at 40 ºC for 3 hours, appears to be the best conditions to obtain the best results of protein extraction, and antioxidant and anti-hypertensive properties. With these conditions, the obtained extracts showed 48% of protein content, an antioxidant activity of 821 μmol TE / g of extract, and an ability of inhibiting the activity of ACE in 61% (using a concentration of 10 mg / mL). Thus, the factorial design allowed to confirm the combination of experimental factors that leads to the most efficient extraction of antioxidant and anti-hypertensive peptides of the mussel Mytilus galloprovincialis. In conclusion, the use of discarded mussels to produce functional ingredients for food, cosmetic and pharmaceutic industries contribute to valorise world waste in a circular economy context.

2021-05Conference object

Open access

Production, characterization and ultrafiltration of bioactive extracts derived from the microalgae Senedesmus obliquus

Publication . Cunha, S. A.; Castro, R. de; Coscueta, E. R.; Silva, J. L. da; Pintado, M. E.

Algae are very abundant worldwide and contains compounds with high biological value, such as proteins, amino acids, antioxidants. It is noteworthy that some algae species contain protein levels similar to meat, soybean and milk. Micro and macroalgae have been studied as a source of bioactive compounds, such as bioactive peptides, with properties of interest for humans and animals. In vitro, enzymatic hydrolysis is one of the most described methods for producing peptides. Thus, in this work the microalgae Senedesmus obliquus was used with the goal of producing water soluble extracts rich in proteins and bioactive peptides. The Senedesmus obliquus used showed 45.7% of protein, 9.1% of lipids, 15.6% of carbohydrates and 15.8% of fibers. Extracts were produced from the microalgae by enzymatic hydrolysis, using ultrapure water as the solvent. First, the mixture was incubated at 50ºC for 2h with a cellulase to weaken the cellular wall and help releasing proteins. Secondly, the mixture was incubated with a protease at 40ºC for 2h, to hydrolyse the proteins into smaller peptides. One half of the produced extract was fractionated by ultrafiltration in a Tangential Flow Filtration System (Cogent μScale) using a cut-off of 3KDa. The protein content was determined by Kjeldahl, the antioxidant activity was determined by ORAC and ABTS assays and anti-hypertensive property was determined by the inhibition method of Angiotensin-I converting enzyme (iACE). The hydrolysis strategy resulted in high levels of hydrolysis with a yield of 52% of soluble protein. The biological activity of non-fractionated extract showed ORAC and iACE results of 530 μmol TE/g of extract and an IC of 200 μg protein/mL, respectively. The showed ORAC results of 597 μmol TE/g of extract. So, due to its protein richness and interesting bioactivities, Senedesmus obliquus extracts may have potential to be used in food and cosmetic industries.

2021-06Conference object

Open access

Valorisation of mussel mytilus galloprovincialis meat waste to produce bioactive extracts by enzymatic hydrolysis

Publication . Cunha, Sara; Castro, Rita de; Coscueta, Ezequiel; Pintado, Manuela

Mytilus galloprovincialis is a mussel consumed and appreciated in several countries. However, its commercialization is associated with waste generation since they are submitted to a preselection before being delivered for sale. This results to the rejection of broken mussels or mussels with size out of the established criteria in the target market. Mussel meat is rich in proteins and has been described as a source of bioactive peptides with interesting properties for food and cosmetic industries. In this work small sized or broken mussels were submitted to enzymatic hydrolysis with a subtilisin protease, testing different conditions to maximize the production of an extract rich in proteins and bioactive peptides. First, the mussel meat was homogenised in a mincer and then submitted to enzymatic hydrolysis with subtilisin, using different combinations of temperature, enzyme concentration and incubation time. The different combinations were stablished using a Box-Behnken experimental design, and their efficacy was achieved by analysing the effect of the three factors on protein release, antioxidant and anti-hypertensive properties. The protein content of each extract was determined by Kjeldahl, the antioxidant activity by oxygen-radical absorbance-capacity (ORAC) assay and anti-hypertensive property by the inhibition method of Angiotensin-I converting enzyme (iACE). The experimental design results were evaluated using statgraphic centurion software. The optimal extraction conditions achieved were the incubation of homogenised mussel meat with 1.5% of enzyme at 52ºC for 3 hours. A scale up extraction was made using the optimized conditions and the resulting extract showed a protein content of 45%, an antioxidant activity of 426 μmol TE/g of extract and the ability inhibiting ACE with an IC50 of 1 mg of protein / mL. Thus, the use of discarded mussels to produce functional ingredients for food, cosmetic and pharmaceutic industries may contribute to valorise world waste in a circular economy context.

2021-06Conference object

Open access

Production, characterization and ultrafiltration of bioactive extracts derived from the microalgae senedesmus obliquus

Publication . Cunha, Sara A.; de Castro, Rita; Coscueta, Ezequiel; Silva, Joana L.; Pintado, Manuela

2021Conference object

Open access