European Food Research and Technology

, Volume 245, Issue 3, pp 535–544 | Cite as

Purification and identification of peptides with high angiotensin-I converting enzyme (ACE) inhibitory activity from honeybee pupae (Apis mellifera) hydrolysates with in silico gastrointestinal digestion

  • Xiaoxue Yang
  • Kangni Chen
  • Huaigao Liu
  • Yuqi Zhang
  • Yongkang LuoEmail author
Original paper


The objective was to identify peptides with ACE inhibitory activity generated from honeybee pupae hydrolysates. In addition, we simulated gastrointestinal digestion and predicted the potential cleavage sites of the identified peptides. Peptides derived from honeybee pupae were cleaved in silico by pepsin, trypsin, and chymotrypsin using the PeptideCutter tool of ExPASy. Honeybee pupae were hydrolyzed using four different proteases (Alcalase, neutrase, trypsin, and papain), and their ACE inhibitory activity and distribution of molecular weights were determined. The neutrase hydrolysates showed the highest ACE inhibitory activity. Then, the hydrolysates were isolated and purified using ultrafiltration, gel filtration chromatography, and reversed-phased high performance liquid chromatography. Three novel ACE inhibitory peptides were identified by LC–MS/MS, AVFPSIVGR, PPVLVFV, and PGKVHIT, which exhibited the most potent ACE inhibitory activity, with IC50 values of 6.64, 47.7859, and 223.869 µM, respectively. In simulated in silico gastrointestinal digestion, peptides AVPFSIVGR and PGKVHIT had the position of cleavage sites by pepsin, chymotrypsin and trypsin, and PPVLVFV was cleaved by pepsin and chymotrypsin. We hope these results support the application of honeybee pupa hydrolysates as functional ingredients in foods and pharmaceuticals.


Honeybee pupae Enzymatic hydrolysates ACE inhibitory activity Functional food Gastrointestinal digestion 



This study was financially supported by the National Science and Technology Ministry of China (Award number 2017YFD0400200).

Compliance with ethical standards

Conflict of interest

We declare no conflicts of interest exist in the submission of this manuscript.

Compliance with ethics requirements

This article does not contain any studies with human or animal subjects.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science and Nutritional EngineeringChina Agricultural UniversityBeijingChina
  2. 2.Beijing Guotai Biotechnology Co., LtdBeijingChina

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