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Purification and characterization of a 6.5 kDa antioxidant peptidoglycan purified from silk worm (Bombyx mori) pupae extract

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Abstract

A unique antioxidant peptidoglycan of 6.5 kDa was purified from silk worm (Bombyx mori) pupae. The peptidoglycan was extracted and isolated from the acidified aqueous extract of the homogenized pupae by chloroform/methanol (2:1, v/v), and purified by size-exclusion chromatography and reverse-phase high-performance liquid chromatography (RP-HPLC). This separation method allowed an easy and consistent preparation of the peptidoglycan from the high protein load by using only 3 steps of purification. The highly pure peptidoglycan consisted of fucose, glucosamine, galactosamine, glucose, and galactose as constituent sugars when revealed by high-performance anion exchange chromatography (HPAEC) analysis. The amino acid composition analysis showed the presence of acidic and polar amino acids (51.7%), hydrophobic (37.2%), and basic amino acids (9.3%) in the purified peptidoglycan. The IC50 of the purified peptidoglycan was 0.22±0.05 and 0.61±0.09 μg for diphenyl-2-picryl hydrazyl (DPPH) and 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), respectively, as quantified by Ophthalaldehyde (OPA) microplate fluorescent assay.

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  1. Department of Food Science & Technology, BK21 Project Team, and Carbohydrate Bioproduct Research Center, Sejong University, Seoul, 143-747, Korea

    Rehab Ahmed Al-Azzouny, Ren Wang & Sang-Ho Yoo

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  1. Rehab Ahmed Al-Azzouny
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  2. Ren Wang
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  3. Sang-Ho Yoo
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Correspondence to Sang-Ho Yoo.

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Al-Azzouny, R.A., Wang, R. & Yoo, SH. Purification and characterization of a 6.5 kDa antioxidant peptidoglycan purified from silk worm (Bombyx mori) pupae extract. Food Sci Biotechnol 20, 243–249 (2011). https://doi.org/10.1007/s10068-011-0033-6

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  • DOI: https://doi.org/10.1007/s10068-011-0033-6

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