Phylogenetic, Structural, and Immunogenic Analysis of Algal L-Asparaginases: Potential Alternatives for ALL Treatment

Document Type : Original Article

Authors

1 Department of Microbiology and Microbial Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran

2 Microbiology and microbial biotechnology department, faculty of life sciences and biotechnology, Shahid Beheshti University

Abstract

L-asparaginase is an essential drug used in the treatment of acute lymphoblastic leukemia (ALL), a highly prevalent cancer in children. However, its immunogenicity and allergenicity often lead to adverse reactions in patients. As a result, alternative strategies, such as identifying novel enzyme sources and employing protein engineering, have been explored. This study aimed to elucidate the molecular structure and predict the immunogenic profile of L-asparaginases from Spirulina subsalsa, Nannochloropsis gaditana CCMP526, and Gracilaria domingensis to propose potential substitutes for bacterial asparaginases, such as EcAII and ErAII. The corresponding enzyme sequences were retrieved from GenBank. Phylogenetic analysis revealed three distinct clusters corresponding to class 1,2, and 3 L-asparaginases. Algal enzymes from N. gaditana CCMP526 (GenBank ID: EWM28374.1) and G. domingensis (GenBank ID: KAI0567449.1) clustered within class 2 L-asparaginases, whereas S. subsalsa (NCBI Reference Sequence ID: WP_265263300.1) exhibited a closer evolutionary relationship with class 3 asparaginases. These findings suggest that algal asparaginases possess unique functional characteristics compared to their bacterial counterparts. Immunogenicity assessment indicated that the T-cell and B-cell epitope densities of S. subsalsa, N. gaditana CCMP526, and G. domingensis were comparable to those of EcAII but significantly lower than ErAII. Additionally, these algal asparaginases demonstrated lower epitope density for the HLA-DRB107:01 allele, which is associated with hypersensitivity reactions, suggesting a reduced likelihood of triggering immune responses. Among the algal sources, N. gaditana exhibited the lowest epitope density, followed by G. domingensis and S. subsalsa. However, in the B-cell epitope analysis, S. subsalsa demonstrated the least potential to elicit allergenic reactions, as it contained only one allergenic epitope. Structural modeling using AlphaFold 3 predicted highly reliable three-dimensional models for the algal asparaginases.

Keywords

References

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Plant, Algae, and Environment
Volume 9, Issue 3
DOi
September 2025
Pages 82-93

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