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The Neutral Protease Immobilization: Physical Characterization of Sodium Alginate-Chitosan Gel Beads

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Abstract

Sodium alginate and chitosan were cross-linked to form composite gel spheres, which were entrapped to immobilize the free neutral protease. The matrix of the immobilized neutral protease was detected and characterized by using Fourier transform infrared spectroscopy and energy-dispersive X-ray. The optimum immobilization conditions were determined by orthogonal test, in which the concentration of sodium alginate was 3.5%, CaCl2 2.5%, chitosan 2.5%, and immobilizing time 1.5 h. Meanwhile, the activities of immobilized neutral protease and free enzyme were compared. The results showed that the pH value of immobilized enzyme was 5–8, the relative activity was above 90%, the free enzyme was above 80%, the relative activity of immobilized enzyme was above 80% in 30–80 °C, and the free enzyme was above 64% in 40–80 °C. The immobilized enzyme is better than the free enzyme in the constant of pH and temperature. The relative activity of immobilized enzyme was 50% after six hydrolysis cycles, and 80% after 11 days of storage.

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Data Availability

Some or all data, models, or code that support the findings of this study are available from the corresponding author upon reasonable request.

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Funding

This research is supported by fund of the Industrial Support Plan Project of Gansu Province colleges and universities, No. 2020C-38.

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Authors and Affiliations

  1. School of Environment and Municipal Engineering, Lanzhou Jiaotong University, 730070, Lanzhou, People’s Republic of China

    Yuan Bai & Wei Wu

  2. Key Laboratory of Yellow River Water Environment in Gansu Province, 730070, Lanzhou, People’s Republic of China

    Yuan Bai

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  1. Yuan Bai
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  2. Wei Wu
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Correspondence to Yuan Bai.

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Highlights

• Neutral protease was immobilized on sodium alginate-chitosan beads, which are promising alternatives to improve the efficiency in enzyme immobilization.

• The optimal conditions for the preparation for immobilized neutral protease were obtained by single-factor optimization and orthogonal methods.

• Sodium alginate-chitosan gel beads have regular spherical structure, good thermal stability and reactivity. The relative enzyme activity remained 50% after 6 cycles

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Bai, Y., Wu, W. The Neutral Protease Immobilization: Physical Characterization of Sodium Alginate-Chitosan Gel Beads. Appl Biochem Biotechnol 194, 2269–2283 (2022). https://doi.org/10.1007/s12010-021-03773-9

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  • DOI: https://doi.org/10.1007/s12010-021-03773-9

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