Exploring β-Glucan from Fungal Isolates as Immunostimulant: Extraction, Characterization, and Medical Applications

Summary

Research Plan

1. Isolation of fungi and yeast from different sources [marine environment "water and sediment" of the Red sea (Suez, Hurghada, Ain Sokhna), date and soil] on different types of media: Wickerham's medium and Potato Dextrose Agar (PDA).

2. Assessment of pathogenicity of the fungal isolates using blood agar medium.

3. Morphological identification (micromorphology) of the fungal isolates using slide culture technique and Dalmau plate technique.

4. Molecular identification of the fungal isolates.

5. Extraction of β-Glucan using alkaline-acidic procedure (AA).

4.6 Purification of β-Glucan using ultracentrifuge.

7. Characterization of β-Glucan using FT-IR spectroscopic analysis and HPLC analysis.

8. Optimization of physical and chemical cultivation conditions using Software (Box-Behnken).

9. Molecular Docking.

10. Medical applications of β-Glucan:

10.1Assessment of In-Vivo Immunomodulatory Effect:

Experimental animal: Mice.

The mice (3 groups, n = 20/group): (G1) a control group, (G2) injected intraperitoneally with cyclosporine, (G3) a group injected intraperitoneally with cyclosporine and beta-glucan. (G2) Each mice will be injected intraperitoneally with 50 mg/kg. (G3) Each mice will be injected intraperitoneally with cyclosporine 50 mg/kg and then will be injected with 150 µg of beta-glucan daily. Every 7 days, mice from G1, G2, and G3 will be sacrificed, and the level of representative cytokines such as IL-6 and IL-10 will be determined in serum by ELISA.

10.2 Antioxidant Activity using ABTS assay
[2, 2' azino-bis (3-ethylbenzothiazoline-6-sulphonic acid)].
10.3 Antitumor Effect using MTT assay.
[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide].
10.4 Antimicrobial Activity of β-Glucan using Well Diffusion Method.

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