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Purification, Identification and Functional Analysis of a Novel Immunomodulatory Peptide from Silkworm Pupa Protein

  • Zhiyong Li
  • Shan Zhao
  • Xiangdong Xin
  • Bei Zhang
  • Attaribo Thomas
  • Asakiya Charles
  • Kwang Sik Lee
  • Byung Rae Jin
  • Zhongzheng GuiEmail author
Article
  • 18 Downloads

Abstract

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.

Keywords

Silkworm pupa protein Immunomodulatory peptide Purification Identification 

Notes

Funding

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.

Ethical approval

This article does not contain any studies with animals performed by any of the authors.

Informed Consent

Informed consent was obtained from all individual participants included in the study.

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Zhiyong Li
    • 1
  • Shan Zhao
    • 1
  • Xiangdong Xin
    • 1
  • Bei Zhang
    • 1
  • Attaribo Thomas
    • 1
  • Asakiya Charles
    • 1
  • Kwang Sik Lee
    • 2
  • Byung Rae Jin
    • 2
  • Zhongzheng Gui
    • 1
    • 3
    Email author
  1. 1.School of BiotechnologyJiangsu University of Science and TechnologyZhenjiangChina
  2. 2.College of Natural Resources and Life ScienceDong-A UniversityBusanSouth Korea
  3. 3.Sericultural Research InstituteChinese Academy of Agricultural ScienceZhenjiangChina

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