Kinetic characterization of two neuraminic acid synthases and evaluation of their application potential

Abstract:

Neuraminic acid synthases are an important yet underexplored group of enzymes. Thus, in this research, we performed a detailed kinetic and stability analysis and a comparison of previously known neuraminic acid synthase from Neisseria meningitidis, and a novel enzyme, PNH5, obtained from a metagenomic library. A systematic analysis revealed a high level of similarity of PNH5 to other known neuraminic acid synthases, except for its pH optimum, which was found to be at 5.5 for the novel enzyme. This is the first reported enzyme from this family that prefers an acidic pH value. The effect of different metal cofactors on enzyme activity, i.e. Co2+, Mn2+ and Mg2+, was studied systematically. The kinetics of neuraminic acid synthesis was completely elucidated, and an appropriate kinetic model was proposed. Enzyme stability study revealed that the purified enzyme exhibits changes in its structure during time as observed by differential light scattering, which cause a drop in its activity and protein concentration. The operational enzyme stability for the neuraminic acid synthase from N. meningitidis is excellent, where no activity drop was observed during the batch reactor experiments. In the case of PNH5, some activity drop was observed at higher concentration of substrates. The obtained results present a solid platform for the future application of these enzymes in the synthesis of sialic acids.

SEEK ID: https://fairdomhub.org/publications/706

DOI: 10.1007/s00253-024-13277-1

Projects: CC-TOP

Publication type: Journal

Journal: Applied Microbiology and Biotechnology

Citation: Çakar, M.M., Milčić, N., Andreadaki, T., Charnock, S., Fessner, W.-D., Findrik Blažević, Z., Kinetic characterization of two neuraminic acid synthases and evaluation of their application potential. Appl Microbiol Biotechnol 108, 446 (2024). https://doi.org/10.1007/s00253-024-13277-1

Date Published: 21st Aug 2024

URL: https://link.springer.com/article/10.1007/s00253-024-13277-1

Registered Mode: manually

Citation
Çakar, M. M., Milčić, N., Andreadaki, T., Charnock, S., Fessner, W.-D., & Blažević, Z. F. (2024). Kinetic characterization of two neuraminic acid synthases and evaluation of their application potential. In Applied Microbiology and Biotechnology (Vol. 108, Issue 1). Springer Science and Business Media LLC. https://doi.org/10.1007/s00253-024-13277-1
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Created: 1st Oct 2024 at 16:59

Last updated: 1st Oct 2024 at 17:12

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