Purification, Identification and Functional Analysis of a Novel Immunomodulatory Peptide from Silkworm Pupa Protein
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In this study, we isolated and characterized an immunomodulatory peptide from silkworm (Bombyx mori) pupa protein hydrolysates. Ultrasound-pretreated hydrolysates were prepared by alcalase digestion and their pro-proliferative activity was assessed with the splenic lymphocyte proliferation assay. Peptide fractions exhibiting the highest activity were purified by Sephadex-G100 and -G15 gel filtration chromatography. The structure of the purified peptides was analyzed using Liquid Chromatography Electrospray Ionisation Tandem Mass Spectrometry (LC–ESI–MS/MS); their digestive stability and their effects of the expression on immune-related cytokines were also evaluated. We purified a novel immunomodulatory peptide with a molecular weight of about 441.06 Da. The amino acid sequence was Asp-His-Ala-Val (DHAV). The splenocyte proliferation rate was 91.1% (P < 0.05) in the presence of 100 µg/mL purified peptide. DHAV was stable in the presence of the gastrointestinal proteases pepsin and trypsin, and stimulated the expression of immune-related factors including interleukin-6 and -12, nuclear factor-κB, cyclin D1, and cyclin-dependent kinase 4. Thus, DHAV purified from silkworm pupa protein hydrolysate has immunomodulatory activity and potential therapeutic value.
KeywordsSilkworm pupa protein Immunomodulatory peptide Purification Identification
This study was funded by the Special Fund for Agro-scientific Research in the Public Interest of China (No. 201403064).
Compliance with Ethical Standards
Conflict of interest
All authors declare that they have no conflict of interest.
This article does not contain any studies with animals performed by any of the authors.
Informed consent was obtained from all individual participants included in the study.
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