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Immobilization of endoglucanase Cel9A on chitosan nanoparticles leads to its stabilization against organic solvents: the use of polyols to improve the stability

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Abstract

The immobilization of enzymes improves their stability in non-conventional media such as organic solvents. In this work, the effects of solvents (DMSO, methanol, ethanol, and n-propanol) on the endoglucanase Cel9A activity and stability were studied. Then, the enzymes were stabilized by its immobilization on chitosan nanoparticles and also using polyols (sorbitol and glycerol) against organic solvents. The SEM results illustrated that the chitosan nanoparticles had about 40 nm diameter. The results indicated that the organic solvents, especially n-propanol, decreased the activity of the free and immobilized enzymes. The reduced activity of the immobilized enzyme was less than that of the free enzyme. Our studies about the enzymes’ stability showed that the free and immobilized enzymes in hydrophobic solvents (with high log P) had the lowest stability compared to other solvents as we observed the half-life of the free enzyme in n-propanol solvent was 2.84 min, and the half-life of the immobilized enzyme was 4.98 min in n-propanol and ethanol solvents 4.50 min. Analysis of the combinatory effects of polyols (sorbitol and glycerol) and the solvents on the stability revealed that sorbitol and glycerol had the most stabilizing effect on the free enzyme in hydrophilic (DMSO) and hydrophobic (n-propanol) solvents, respectively. However, the stabilizing effects of polyols in the immobilized enzyme were independent of the solvents’ hydrophobicity (or log P) due to the hydrophilic properties of chitosan nanoparticles. Therefore, one can conclude that the physiochemical properties of nanoparticles (such as hydrophilicity) influence the stabilizing effects of polyols on immobilized enzyme.

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Abbreviations

AaCel9A:

Endoglucanase Cel9A from Alicyclobacillus acidocaldarius

ChNPs:

Chitosan nanoparticles

DMSO:

Dimethyl sulfoxide

DNS:

Dinitrosalcylic acid

CMC:

Carboxymethyl cellulose

TPP:

Tripolyphosphate

SEM:

Scanning electron microscopy

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Authors and Affiliations

  1. Department of Cellular and Molecular Biology, Faculty of Science, Azarbaijan Shahid Madani University, Tabriz, Iran

    Masoumeh Mohammadi, Saeed Najavand & Mohammad Pazhang

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  1. Masoumeh Mohammadi
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  2. Saeed Najavand
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  3. Mohammad Pazhang
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Correspondence to Saeed Najavand.

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Mohammadi, M., Najavand, S. & Pazhang, M. Immobilization of endoglucanase Cel9A on chitosan nanoparticles leads to its stabilization against organic solvents: the use of polyols to improve the stability. 3 Biotech 9, 269 (2019). https://doi.org/10.1007/s13205-019-1794-5

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