European Journal of Biology

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Research Article


DOI :10.26650/EurJBiol.2024.1454771   IUP :10.26650/EurJBiol.2024.1454771    Full Text (PDF)

Investigating the Antioxidant Capacity of Lunasin Expressed in Aspergillus oryzae

Elif Karaman, Cem Albayrak, Serdar Uysal

Objective: Lunasin is a bioactive protein that possesses anti-carcinogenic, anti-inflammatory, and antioxidant properties. Traditional isolation methods are resource-intensive, and chemical synthesis faces cost and environmental issues. This study aims to achieve cost-effective lunasin expression in Aspergillus oryzae with a focus on exploring its antioxidant properties in vitro.

Materials and Methods: The expression vector carrying four lunasin sequences fused with amylase and an 8xHis-tag was introduced into pyrG auxotrophic A. oryzae. Subsequently, the recombinant protein was purified using metal affinity chromatography. The study uses sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), western blot analyses, and size-exclusion chromatography to evaluate the composition and purity of the protein, a linoleic acid assay to demonstrate the inhibitory effect on lipid peroxidation, and the 2,2’-azinobis-[3-ethylbenzothiazoline-6-sulfonic acid] ABTS) assay to evaluate the radical scavenging activity.

Results: SDS-PAGE and western blot analyses confirmed sustained lunasin expression in A. oryzae, appearing in both fusion and non-fusion forms. Yields were 5.8 mg/L for non-fusion and 4 mg/L for fusion lunasin expression. Moreover, 0.1 μM non-fusion lunasin surpassed α-tocopherol and butylated hydroxyanisole (BHA; p < 0.05) in reducing lipid peroxidation at 4 and 72 h. Unlike the fusion lunasin, the non-fusion lunasin displayed concentration- and time-independent inhibitory effects on linoleic acid peroxidation as well as significant ABTS scavenging activity (p < 0.05).

Conclusion: The study has shown for the first time A. oryzae to efficiently express and secrete both fusion and non-fusion lunasin proteins in a soluble form, with the non-fusion lunasin exhibiting superior antioxidant effectiveness compared to the fusion lunasin. The findings underscore A. oryzae’s potential as a promising host for producing functional lunasin with antioxidant properties, opening avenues for broader applications in biotechnology and bioactive peptides.

Keywords: Lunasin, Aspergillus oryzae, Antioxidant, Biotechnology

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APA

Karaman, E., Albayrak, C., & Uysal, S. (2024). Investigating the Antioxidant Capacity of Lunasin Expressed in Aspergillus oryzae. European Journal of Biology, 83(1), 67-76. https://doi.org/10.26650/EurJBiol.2024.1454771


AMA

Karaman E, Albayrak C, Uysal S. Investigating the Antioxidant Capacity of Lunasin Expressed in Aspergillus oryzae. European Journal of Biology. 2024;83(1):67-76. https://doi.org/10.26650/EurJBiol.2024.1454771


ABNT

Karaman, E.; Albayrak, C.; Uysal, S. Investigating the Antioxidant Capacity of Lunasin Expressed in Aspergillus oryzae. European Journal of Biology, [Publisher Location], v. 83, n. 1, p. 67-76, 2024.


Chicago: Author-Date Style

Karaman, Elif, and Cem Albayrak and Serdar Uysal. 2024. “Investigating the Antioxidant Capacity of Lunasin Expressed in Aspergillus oryzae.” European Journal of Biology 83, no. 1: 67-76. https://doi.org/10.26650/EurJBiol.2024.1454771


Chicago: Humanities Style

Karaman, Elif, and Cem Albayrak and Serdar Uysal. “Investigating the Antioxidant Capacity of Lunasin Expressed in Aspergillus oryzae.” European Journal of Biology 83, no. 1 (Nov. 2024): 67-76. https://doi.org/10.26650/EurJBiol.2024.1454771


Harvard: Australian Style

Karaman, E & Albayrak, C & Uysal, S 2024, 'Investigating the Antioxidant Capacity of Lunasin Expressed in Aspergillus oryzae', European Journal of Biology, vol. 83, no. 1, pp. 67-76, viewed 8 Nov. 2024, https://doi.org/10.26650/EurJBiol.2024.1454771


Harvard: Author-Date Style

Karaman, E. and Albayrak, C. and Uysal, S. (2024) ‘Investigating the Antioxidant Capacity of Lunasin Expressed in Aspergillus oryzae’, European Journal of Biology, 83(1), pp. 67-76. https://doi.org/10.26650/EurJBiol.2024.1454771 (8 Nov. 2024).


MLA

Karaman, Elif, and Cem Albayrak and Serdar Uysal. “Investigating the Antioxidant Capacity of Lunasin Expressed in Aspergillus oryzae.” European Journal of Biology, vol. 83, no. 1, 2024, pp. 67-76. [Database Container], https://doi.org/10.26650/EurJBiol.2024.1454771


Vancouver

Karaman E, Albayrak C, Uysal S. Investigating the Antioxidant Capacity of Lunasin Expressed in Aspergillus oryzae. European Journal of Biology [Internet]. 8 Nov. 2024 [cited 8 Nov. 2024];83(1):67-76. Available from: https://doi.org/10.26650/EurJBiol.2024.1454771 doi: 10.26650/EurJBiol.2024.1454771


ISNAD

Karaman, Elif - Albayrak, Cem - Uysal, Serdar. “Investigating the Antioxidant Capacity of Lunasin Expressed in Aspergillus oryzae”. European Journal of Biology 83/1 (Nov. 2024): 67-76. https://doi.org/10.26650/EurJBiol.2024.1454771



TIMELINE


Submitted21.03.2024
Accepted30.04.2024
Published Online30.05.2024

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