Biotechnological Potential of Rheinheimera sp. L-asparaginase: Heterologous Production and Its Role in Acrylamide Mitigation

Yıl 2025, Cilt: 15 Sayı: 1, 330 - 342, 01.03.2025

Ahmet Tülek

https://doi.org/10.21597/jist.1526603

Öz

Acrylamide, a chemical contaminant found in foods, poses a significant health threat because of its toxic and carcinogenic properties. One of the most effective methods for reducing acrylamide is the application of L-asparaginase (L-ASNase) to decrease the asparagine content in foods before cooking or processing. In this study, L-ASNase (RsASNase) from Rheinheimera sp. was expressed heterologously in Escherichia coli Rosetta™2 (DE3) host cells. The enzyme was purified using Ni2+-NTA affinity chromatography, yielding a specific activity of 392.2 U/mg and a purification fold of 4.0. Acrylamide reduction was assessed using a starch-L-asparagine model analyzed by high-performance liquid chromatography (HPLC). The highest acrylamide mitigation (52.3%) was achieved using 100 U of the enzyme after 120 min of incubation. Additionally, the three-dimensional structure of RsASNase was modeled using the ProMod3. Bioinformatics analyses, including docking studies, revealed interactions between the RsASNase enzyme's active site and the L-asparagine substrate, involving the amino acids THR162A, LYS242A, THR273A, LEU304A, and GLU305A. These findings showed that RsASNase has the potential for further development and application in biotechnological processes aimed at acrylamide mitigation.

Anahtar Kelimeler

L-asparaginase, Rheinheimera sp., Acrylamide mitigation, Heterologous expression

Kaynakça

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