Improving protein extraction and peptide production from Chlorella vulgaris using combined mechanical/physical and enzymatic pre-treatments
Open accessAbstract
Chlorella vulgaris is a microalga rich in proteins with potential applications in food and feed industries. However, the presence of a cellulose-containing cell wall, which is a major barrier to protein extraction, together with fibroproteinaceous complexes, limits the bioaccessibility of nutritional and bioactive proteins and peptides from C. vulgaris biomass. Therefore, this study aimed to evaluate the effect of different mechanical/physical pre-treatments (bead milling, extrusion, freeze-drying, heating, microwave and sonication) combined or not with enzymatic treatments (commercial trypsin and pancreatin) on protein extraction and peptide formation from a C. vulgaris suspension. The amount of total protein and peptides released to the supernatant was quantified by Bradford and o -phthaldialdehyde assays, respectively. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis was used to analyse the extracted protein fractions. The results showed that extrusion caused a 3-fold increase in total peptides (p < 0.001) compared to no-pretreatment, and trypsin increased peptides formed in bead-milled (p = 0.020) and freeze-dried (p = 0.021) microalga relative to those pre-treatments alone. Some pre-treatments, such as bead milling and microwave, were effective in releasing specific protein fractions, particularly those from 32 to 40 kDa (up to 1.2-fold), compared to control. Pancreatin combined with bead milling decreased 32 to 40 kDa- and 26 kDa-protein fractions (p < 0.010) compared with the sole use of mechanical treatment, whereas the same enzyme mixture associated with microwave produced a similar result for 26 kDa-protein fraction (p = 0.023). Pancreatin also effectively reduced the total protein fraction released after pre-treatment with sonication (p = 0.013). These findings suggest that combining different pre-treatments and enzymatic treatments could improve protein extraction from C. vulgaris biomass, providing a useful approach for the development of sustainable protein sources. The present results highlight the need for further studies to assess the efficacy of extrusion in improving the bioaccessibility of C. vulgaris proteins in monogastric animals' diets.
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The presence of amino acids such as methionine, phenylalanine, threonine, and valine in microalgal proteins enhances nutritional value and influences conformational stability and intermolecular interaction capacity, improving thermal stability and technological functionality.
“methionine, phenylalanine, threonine, and valine, not only enhances nutritional value but also influences conformational stability and intermolecular interaction capacity, resulting in improved thermal stability and technological functionality (Costa et al., 2024; Wang et al., 2024).”
From Microalgae biotechnology and its role in sustainable and healthy food design (2025) · cites this paper as Costa et al., 2024
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- Authors
- Costa, M. M., Spínola, M. P., Alves, V. D., & Mestre Prates, J. A.
- Affiliations
- University of Lisbon (PT)
- Venue
- Heliyon
- Publisher
- Elsevier BV
- Published
- 2024-06
- Volume
- 10
- Issue
- 12
- Pages
- e32704
- ISSN
- 2405-8440 (ISSN-L)
- PMC
- PMC11233943
- OpenAlex
- W4399421937
- Cited by
- 40
- References
- 43
- Citations by year
- 2026: 17 · 2025: 20 · 2024: 3
- FWCI
- 5.45
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Heliyon (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- Funders
- Fundação para a Ciência e a Tecnologia (UI/BD/153071/2022, UIDB/00276/2020, LA/P/0059/2020, P2020/17/SI/70114/2019)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Hydrolysis and Bioactive Peptides
- Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Algal biology and biofuel production
- Life Sciences › Agricultural and Biological Sciences › Aquatic Science › Seaweed-derived Bioactive Compounds
- Keywords
- Chlorella vulgaris, bioactive peptides, enzymatic pretreatment, trypsin, pancreatin, bead milling, extrusion, freeze-drying, microwave pretreatment, sonication, cell wall, protein bioaccessibility, sustainable protein sources, microalgal biomass
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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