Advertisement

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
  • 23 Downloads

Abstract

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.

Keywords

Honeybee pupae Enzymatic hydrolysates ACE inhibitory activity Functional food Gastrointestinal digestion 

Notes

Acknowledgements

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.

References

  1. 1.
    Sun-Waterhouse D, Waterhouse G, You L, Zhang J, Liu Y, Ma L, Gao J, Dong Y (2016) Transforming insect biomass into consumer wellness foods: a review. Food Res Int 89:129–151CrossRefGoogle Scholar
  2. 2.
    Nongonierma AB, FitzGerald RJ (2017) Unlocking the biological potential of proteins from edible insects through enzymatic hydrolysis: a review. Innov Food Sci Emerg 43:239–252CrossRefGoogle Scholar
  3. 3.
    Lautenschläger T, Neinhuis C, Monizi M, Mandombe JL, Förster A, Henle T, Nuss M (2017) Edible insects of Northern Angola. Afr Invertebr 58:55–82CrossRefGoogle Scholar
  4. 4.
    Belluco S, Losasso C, Maggioletti M, Alonzi CC, Paoletti MG, Ricci A (2013) Edible insects in a food safety and nutritional perspective: a critical review. Compr Rev Food Sci Food Saf 12:296–313CrossRefGoogle Scholar
  5. 5.
    Veldkamp T, Bosch G (2015) Insects: a protein-rich feed ingredient in pig and poultry diets. Anim Front 5:45–50Google Scholar
  6. 6.
    Shrikant S, Raghvendar S, Shashank R (2011) Bioactive peptides: a review. Int J Bioautomation 15:223–250Google Scholar
  7. 7.
    Vercruysse L, Smagghe G, Matsui T, Van Camp J (2008) Purification and identification of an angiotensin I converting enzyme (ACE) inhibitory peptide from the gastrointestinal hydrolysate of the cotton leafworm, Spodoptera littoralis. Process Biochem 43:900–904CrossRefGoogle Scholar
  8. 8.
    Tao M, Sun H, Liu L, Luo X, Lin G, Li R, Zhao Z, Zhao Z (2017) Graphitized porous carbon for rapid screening of angiotensin-converting enzymeinhibitory peptide GAMVVH from silkworm pupa protein and molecular insight into inhibition mechanism. J Agric Food Chem 65:8626–8633CrossRefGoogle Scholar
  9. 9.
    Dai C, Ma H, Luo L, Yin X (2013) Angiotensin I-converting enzyme (ACE) inhibitory peptide derived from Tenebrio molitor (L.) larva protein hydrolysate. Eur Food Res Technol 236:681–689CrossRefGoogle Scholar
  10. 10.
    Pattarayingsakul W, Nilavongse A, Reamtong O, Chittavanich P, Mungsantisuk I, Mathong Y, Prasitwuttisak W, Panbangred W (2017) Angiotensin-converting enzyme inhibitory and antioxidant peptides from digestion of larvae and pupae of Asian weaver ant, Oecophylla smaragdina, Fabricius. J Sci Food Agric 97(10):3133–3140CrossRefGoogle Scholar
  11. 11.
    Wijesekara I, Kim SK (2010) Angiotensin-I-converting enzyme (ACE) inhibitors from marine resources: prospects in the pharmaceutical industry. Mar Drugs 8:1080–1093CrossRefGoogle Scholar
  12. 12.
    Vaštag Ž, Popović L, Popović S, Krimer V, Peričin D (2011) Production of enzymatic hydrolysates with antioxidant and angiotensin-I converting enzyme inhibitory activity from pumpkin oil cake protein isolate. Food Chem 124:1316–1321CrossRefGoogle Scholar
  13. 13.
    Ghosh S, Jung C, Meyer-Rochow VB (2016) Nutritional value and chemical composition of larvae, pupae, and adults of worker honey bee, Apis mellifera ligustica as a sustainable food source. J Asia-Pac Entomol l19:487–495CrossRefGoogle Scholar
  14. 14.
    Woltedji D, Song F, Zhang L, Gala A, Han B, Feng M, Fang Y, Li J (2012) Western honeybee drones and workers (Apis mellifera ligustica) have different olfactory mechanisms than eastern honeybees (Apis cerana cerana). J Proteome Res 11:4526–4540CrossRefGoogle Scholar
  15. 15.
    Jensen AB, Evans J, Jonas-Levi A, Benjamin O, Itzhak Martinez I, Dahle B, Roos N, Lecocq A, Foley K (2016) Standard methods for Apis mellifera brood as human food. J Apicult Res 56:1–28CrossRefGoogle Scholar
  16. 16.
    Rumpold BA, Schluter OK (2013) Nutritional composition and safety aspects of edible insects. Mol Nutr Food Res 57:802–823CrossRefGoogle Scholar
  17. 17.
    Evans J, Mueller A, Jensen AB, Dahle B, Flore R, Eilenberg J, Frost MB (2016) A descriptive sensory analysis of honeybee drone brood from Denmark and Norway. J Insects Food Feed 2:277–283CrossRefGoogle Scholar
  18. 18.
    Li Y, Jiang B, Zhang T, Mu W, Liu J (2008) Antioxidant and free radical-scavenging activities of chickpea protein hydrolysate (CPH). Food Chem 106:444–450CrossRefGoogle Scholar
  19. 19.
    Xie N, Liu S, Wang C, Li B (2014) Stability of casein antioxidant peptide fractions during in vitro digestion/Caco-2 cell model: characteristics of the resistant peptides. Eur Food Res Technol 239:577–586CrossRefGoogle Scholar
  20. 20.
    Friedland J, Silverstein E (1977) Sensitive fluorimetric assay for serum angiotensin-converting enzyme with natural substrate angiotensin-1. Am J Clin Pathol 68:225–228CrossRefGoogle Scholar
  21. 21.
    Gao D, Cao Y, Mai X (2011) Modified spectrophotometric method for assay of angiotensin I-converting enzyme inhibitory activity of food-derived peptides. J Zhejiang Univ 37:219–223Google Scholar
  22. 22.
    Wu Q, Du J, Jia J, Kuang C (2016) Production of ACE inhibitory peptides from sweet sorghum grain protein using alcalase: hydrolysis kinetic, purification and molecular docking study. Food Chem 199:140–149CrossRefGoogle Scholar
  23. 23.
    Zhang C, Zhang Y, Wang Z, Chen S, Luo Y (2017) Production and identification of antioxidant and angiotensin-converting enzyme inhibition and dipeptidyl peptidase IV inhibitory peptides from bighead carp (Hypophthalmichthys nobilis) muscle hydrolysate. J Funct Foods 35:224–235CrossRefGoogle Scholar
  24. 24.
    Chi C-F, Wang B, Wang Y-M, Zhang B, Deng S-G (2015) Isolation and characterization of three antioxidant peptides from protein hydrolysate of bluefin leatherjacket (Navodon septentrionalis) heads. J Funct Foods 12:1–10CrossRefGoogle Scholar
  25. 25.
    Kim S, Byun H, Park P, Shahidi F (2001) Angiotensin I converting enzyme inhibitory peptides purified from bovine skin gelatin hydrolysate. J Agric Food Chem 49:2992–2996CrossRefGoogle Scholar
  26. 26.
    Chen J, Wang Y, Zhong Q, Wu Y, Xia W (2012) Purification and characterization of a novel angiotensin-I converting enzyme (ACE) inhibitory peptide derived from enzymatic hydrolysate of grass carp protein. Peptides 33:52–58CrossRefGoogle Scholar
  27. 27.
    Vermeirssen V, Van Camp J, Verstraete W (2005) Fractionation of angiotensin I converting enzyme inhibitory activity from pea and whey proteinin vitro gastrointestinal digests. J Sci Food Agric 85:399–405CrossRefGoogle Scholar
  28. 28.
    Megias C, Yust MD, Pedroche J, Lquari H, Giron-Calle J, Alaiz M, Millan F, Vioque J (2004) Purification of an ACE inhibitory peptide after hydrolysis of sunflower (Helianthus annuus L.) protein isolates. J Agric Food Chem 52:1928–1932CrossRefGoogle Scholar
  29. 29.
    Zhang M, Mu T-H, Sun M-J (2014) Purification and identification of antioxidant peptides from sweet potato protein hydrolysates by Alcalase. J Funct Foods 7:191–200CrossRefGoogle Scholar
  30. 30.
    Tao M, Wang C, Liao D, Liu H, Zhao Z, Zhao Z (2017) Purification, modification and inhibition mechanism of angiotensin I-converting enzyme inhibitory peptide from silkworm pupa (Bombyx mori) protein hydrolysate. Process Biochem 54:172–179CrossRefGoogle Scholar
  31. 31.
    Tsai J-S, Chen J-L, Pan BS (2008) ACE-inhibitory peptides identified from the muscle protein hydrolysate of hard clam (Meretrix lusoria). Process Biochem 43:743–747CrossRefGoogle Scholar
  32. 32.
    Lau CC, Abdullah N, Shuib AS, Aminudin N (2014) Novel angiotensin I-converting enzyme inhibitory peptides derived from edible mushroom Agaricus bisporus (J.E. Lange) Imbach identified by LC–MS/MS. Food Chem 148:396–401CrossRefGoogle Scholar
  33. 33.
    Nakamura Y, Yamamoto N, Sakai K, Okubo A, Yamazaki S, Takano T (1995) Purification and characterization of angiotensin I-converting enzyme-inhibitors from sour milk. J Dairy Sci 78:777–783CrossRefGoogle Scholar
  34. 34.
    Garcia-Mora P, Martin-Martinez M, Angeles Bonache M, Gonzalez-Muniz R, Penas E, Frias J, Martinez-Villaluenga C (2017) Identification, functional gastrointestinal stability and molecular docking studies of lentil peptides with dual antioxidant and angiotensin I converting enzyme inhibitory activities. Food Chem 221:464–472CrossRefGoogle Scholar
  35. 35.
    Byun H, Kim S (2001) Purification and characterization of angiotensin I converting enzyme (ACE) inhibitory peptides from Alaska pollack (Theragra chalcogramma) skin. Process Biochem 36:1155–1162CrossRefGoogle Scholar
  36. 36.
    Toopcham T, Mes JJ, Wichers HJ, Roytrakul S, Yongsawatdigul J (2017) Bioavailability of angiotensin I-converting enzyme (ACE) inhibitory peptides derived from Virgibacillus halodenitrificans SK1-3-7 proteinases hydrolyzed tilapia muscle proteins. Food Chem 220:190–197CrossRefGoogle Scholar
  37. 37.
    Fu Y, Young JF, Løkke MM, Lametsch R, Aluko RE, Therkildsen M (2016) Revalorisation of bovine collagen as a potential precursor of angiotensin I-converting enzyme (ACE) inhibitory peptides based on in silico and in vitro protein digestions. J Funct Foods 24:196–206CrossRefGoogle Scholar
  38. 38.
    Udenigwe CC (2014) Bioinformatics approaches, prospects and challenges of food bioactive peptide research. Trends Food Sci Technol 36:137–143CrossRefGoogle Scholar

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

Personalised recommendations