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Exploration of cyclooxygenase-2 inhibitory peptides from walnut dreg proteins based on in silico and in vitro analysis

Zishan Honga,b,c,1, Jing Xiea,b,c,1, Liang Taoa,b,c,1, Jing-Jing Daib,d,e, Tingting Lia,b,e, Li Zhanga,c,e, Yuying Baia,b,e, Xia Hua,c,e, Jinlian Chena,b,e, Jun Shenga,f(), Yang Tiana,b,c,d,e()
College of Food Science and Technology, Yunnan Agricultural University, Kunming 650201, China
Yunnan Provincial Key Laboratory of Precision Nutrition and Personalized Food Manufacturing, Yunnan Agricultural University, Kunming 650201, China
Engineering Research Center of Development and Utilization of Food and Drug Homologous Resources, Ministry of Education, Yunnan Agricultural University, Kunming 650201, China
School of Tea and Coffee, Pu’er University, Pu’er 665000, China
Yunnan Provincial Engineering Research Center for Edible and Medicinal Homologous Functional Food, Yunnan Agricultural University, Kunming 650201, China
Key Laboratory of Pu-er Tea Science, Ministry of Education, Yunnan Agricultural University, Kunming 650201, China

1 These authors contributed equally to this study.

Peer review under responsibility of Tsinghua University Press.

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Highlights

• Peptides in walnut dreg proteins are efficiently released by alcalase.

• Alcalase hydrolysate (Al-WPH) inhibits cyclooxygenase-2 (COX-2) activity.

• Four novel peptides (AGFP, FPGA, LFPD, and VGFP) were identified in Al-WPH.

• AGFP, FPGA, LFPD, and VGFP showed potential COX-2 inhibitory activity.

• Walnut dreg protein is an excellent low-cost source of anti-inflammatory peptides.

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Abstract

Walnut dreg protein hydrolysates (WDPHs) exhibit a variety of biological activities, however, the cyclooxygenase-2 (COX-2) inhibitory peptide of WDPHs remain unclear. The aim of this study was to rapidly screen for such peptides in WDPHs through a combination of in silico and in vitro analysis. In total, 1262 peptide sequences were observed by nano liquid chromatography/tandem mass spectrometry (nano LC-MS/MS) and 4 novel COX-2 inhibitory peptides (AGFP, FPGA, LFPD, and VGFP) were identif ied. Enzyme kinetic data indicated that AGFP, FPGA, and LFPD displayed mixed-type COX-2 inhibition, whereas VGFP was a non-competitive inhibitor. This is mainly because the peptides form hydrogen bonds and hydrophobic interactions with residues in the COX-2 active site. These results demonstrate that computer analysis combined with in vitro evaluation allows for rapid screening of COX-2 inhibitory peptides in walnut protein dregs.

Keywords

Walnut dreg proteinsCyclooxygenase-2 inhibitory peptideIdentificationVirtual screeningMolecular docking

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References

[1]

K. Uchida, HNE as an inducer of COX-2, Free Radic. Biol. Med. 111 (2017) 169-172. https://doi.org/10.1016/j.freeradbiomed.2017.02.004.

Crossref Google Scholar
[2]

M.A.H. Khan, S.H. Hwang, S.D. Barnett, et al., Multitarget molecule, PTUPB, to treat diabetic nephropathy in rats, Br. J. Pharmacol. 178 (2021) 4468-4484. https://doi.org/10.1111/bph.15623.

Crossref Google Scholar
[3]

C. Cozowicz, J. Poeran, N. Zubizarreta, et al., Non-opioid analgesic modes of pain management are associated with reduced postoperative complications and resource utilisation: a retrospective study of obstructive sleep apnoea patients undergoing elective joint arthroplasty, Br. J. Anaesth. 122 (2019) 131-140. https://doi.org/10.1016/j.bja.2018.08.027.

Crossref Google Scholar
[4]

Y.H. Luan, D. Wang, Q. Yu, et al., Action of β-endorphin and nonsteroidal anti-inflammatory drugs, and the possible effects of nonsteroidal antiinflammatory drugs on β-endorphin, J. Clin. Anesth. 37 (2017) 123-128. https://doi.org/10.1016/j.jclinane.2016.12.016.

Crossref Google Scholar
[5]

A. Karateev, A. Tsurgan, N. Gontarenko, AB0987 Recurrence of Nsaid -induced ulcers: Has the situation changed in recent years? Ann. Rheum. Dis. 75 (2016) 1238. https://doi.org/10.1136/annrheumdis-2016-eular.4708.

Crossref Google Scholar
[6]

K.B. Sondergaard, G. Gislason, NSAIDs and cardiac arrest: non-steroidal anti-inflammatory drug use is associated with increased risk of out-ofhospital cardiac arrest: a nationwide case–time–control study, Eur. Heart J. 38 (2017) 1788-1789. https://doi.org/10.1093/ehjcvp/pvw041.

Crossref Google Scholar
[7]

D. Liu, Y. Guo, H. Ma, Production, bioactivities and bioavailability of bioactive peptides derived from walnut origin by-products: a review, Crit. Rev. Food Sci. Nutr. (2022) 1-16. https://doi.org/10.1080/10408398.2022.2054933.

Crossref Google Scholar
[8]

T. Zhi, D. Hong, Z. Zhang, et al., Anti-inflammatory and gut microbiota regulatory effects of walnut protein derived peptide LPF in vivo, Food Res. Int. 152 (2022) 110875. https://doi.org/10.1016/j.foodres.2021.110875.

Crossref Google Scholar
[9]

X. Kong, L. Zhang, W. Song, et al., Separation, identification and molecular binding mechanism of dipeptidyl peptidase IV inhibitory peptides derived from walnut (Juglans regia L.) protein, Food Chem. 347 (2021) 129062. https://doi.org/10.1016/j.foodchem.2021.129062.

Crossref Google Scholar
[10]

D. Liu, Y. Guo, P. Wu, et al., The necessity of walnut proteolysis based on evaluation after in vitro simulated digestion: ACE inhibition and DPPH radical-scavenging activities, Food Chem. 311 (2020) 125960. https://doi.org/10.1016/j.foodchem.2019.125960.

Crossref Google Scholar
[11]

S. Sun, X. Xu, X. Sun, et al., Preparation and identification of ACE inhibitory peptides from the marine macroalga Ulva intestinalis, Mar. Drugs 17 (2019) 179. https://doi.org/10.3390/md17030179.

Crossref Google Scholar
[12]

H. Sun, Q. Chang, L. Liu, et al., High-throughput and rapid screening of novel ACE inhibitory peptides from sericin source and inhibition mechanism by using in silico and in vitro prescriptions, J. Agric. Food Chem. 65 (2017)10020-10028. https://doi.org/10.1021/acs.jafc.7b04043.

Crossref Google Scholar
[13]

C. Yan, Z. Zhou, Walnut pellicle phenolics greatly influence the extraction and structural properties of walnut protein isolates, Food Res. Int. 141 (2021) 110163. https://doi.org/10.1016/j.foodres.2021.110163.

Crossref Google Scholar
[14]

S. Wang, G. Su, J. Fan, et al., Arginine-containing peptides derived from walnut protein against cognitive and memory impairment in scopolamineinduced zebrafish: design, release, and neuroprotection, J. Agric. Food Chem. 70 (2022) 11579-11590. https://doi.org/10.1021/acs.jafc.2c05104.

Crossref Google Scholar
[15]

S. Gupta, P. Kapoor, K. Chaudhary, et al., In silico approach for predicting toxicity of peptides and proteins, PLoS One 8 (2013) e73957. https://doi.org/10.1371/journal.pone.0073957.

Crossref Google Scholar
[16]

Q. Li, C. Zhang, H. Chen, et al., BioPepDB: an integrated data platform for food-derived bioactive peptides, Int. J. Food Sci. Nutr. 69 (2018) 963-968. https://doi.org/10.1080/09637486.2018.1446916.

Crossref Google Scholar
[17]

A.A. Zamyatnin, A.S. Borchikov, M.G. Vladimirov, et al., The EROPMoscow oligopeptide database, Nucleic Acids Res. 34 (2006) D261-D266. https://doi.org/10.1093/nar/gkj008.

Crossref Google Scholar
[18]

H. Zhang, G. Chen, Y. Zhang, et al., Potential hypoglycemic, hypolipidemic, and anti-inflammatory bioactive components in Nelumbo nucifera leaves explored by bioaffinity ultrafiltration with multiple targets, Food Chem. 375 (2022) 131856. https://doi.org/10.1016/j.foodchem.2021.131856.

Crossref Google Scholar
[19]

M. Yan, H. Tao, S. Qin, Effect of enzyme type on the antioxidant activities and functional properties of enzymatic hydrolysates from sea cucumber (Cucumaria frondosa) viscera, J. Aquat. Food Prod. Technol. 25 (2016) 940-952. https://doi.org/10.1080/10498850.2014.994083.

Crossref Google Scholar
[20]

C. Yu, L. Zheng, Y. Cai, et al., Desirable characteristics of casein peptides with simultaneously enhanced emulsion forming ability and antioxidative capacity in O/W emulsion, Food Hydrocoll. 131 (2022) 107812. https://doi.org/10.1016/j.foodhyd.2022.107812.

Crossref Google Scholar
[21]

N. Lapsongphon, J. Yongsawatdigul, Production and purification of antioxidant peptides from a mungbean meal hydrolysate by Virgibacillus sp.SK37 proteinase, Food Chem. 141 (2013) 992-999. https://doi.org/10.1016/j.foodchem.2013.04.054.

Crossref Google Scholar
[22]

M. Merz, T. Eisele, P. Berends, et al., Flavourzyme, an enzyme preparation with industrial relevance: automated nine-step purification and partial characterization of eight enzymes, J. Agric. Food Chem. 63 (2015) 5682-5693. https://doi.org/10.1021/acs.jafc.5b01665.

Crossref Google Scholar
[23]

R. Jin, X. Teng, J. Shang, et al., Identification of novel DPP-IV inhibitory peptides from Atlantic salmon (Salmo salar) skin, Food Res. Int. 133 (2020) 109161. https://doi.org/10.1016/j.foodres.2020.109161.

Crossref Google Scholar
[24]

S. Rawdkuen, N. Rodzi, S. Pinijsuwan, Characterization of sacha inchi protein hydrolysates produced by crude papain and Calotropis proteases, LWT-Food Sci. Technol. 98 (2018) 18-24. https://doi.org/10.1016/j.lwt.2018.08.008.

Crossref Google Scholar
[25]

Z.Y. Li, W. Youravong, A.H. Kittikun, Protein hydrolysis by protease isolated from tuna spleen by membrane filtration: a comparative study with commercial proteases, LWT-Food Sci. Technol. 43 (2010) 166-172. https://doi.org/10.1016/j.lwt.2009.07.002.

Crossref Google Scholar
[26]

S. Wang, L. Zheng, T. Zhao, et al., Inhibitory effects of walnut (Juglans regia) peptides on neuroinflammation and oxidative stress in lipopolysaccharide-induced cognitive impairment mice, J. Agric. Food Chem. 68 (2020) 2381-2392. https://doi.org/10.1021/acs.jafc.9b07670.

Crossref Google Scholar
[27]

S.K. Sharma, B.J. Al-Hourani, M. Wuest, et al., Synthesis and evaluation of fluorobenzoylated di- and tripeptides as inhibitors of cyclooxygenase-2 (COX-2), Bioorg. Med. Chem. 20 (2012) 2221-2226. https://doi.org/10.1016/j.bmc.2012.02.021.

Crossref Google Scholar
[28]

S.A. Nankar, A.H. Pande, Properties of apolipoprotein E derived peptide modulate their lipid-binding capacity and influence their anti-inflammatory function, Biochim. Biophys. Acta 1841 (2014) 620-629. https://doi.org/10.1016/j.bbalip.2014.01.006.

Crossref Google Scholar
[29]

R. Suttisuwan, S. Phunpruch, T. Saisavoey, et al., Isolation and characterization of anti-inflammatory peptides derived from trypsin hydrolysis of microalgae protein (Synechococcus sp. VDW), Food Biotechnol. 33 (2019) 303-324. https://doi.org/10.1080/08905436.2019.1673171.

Crossref Google Scholar
[30]

V.G. Tacias-Pascacio, R. Morellon-Sterling, E.H. Siar, et al., Use of alcalase in the production of bioactive peptides: a review, Int. J. Biol. Macromol. 165 (2020) 2143-2196. https://doi.org/10.1016/j.ijbiomac.2020.10.060.

Crossref Google Scholar
[31]

J. Fernández-Lucas, D. Castañeda, D. Hormigo, New trends for a classical enzyme: papain, a biotechnological success story in the food industry, Trends Food Sci. Technol. 68 (2017) 91-101. https://doi.org/10.1016/j.tifs.2017.08.017.

Crossref Google Scholar
[32]

G. Rajasekaran, R. Kamalakannan, S.Y. Shin, Enhancement of the antiinflammatory activity of temporin-1Tl-derived antimicrobial peptides by tryptophan, arginine and lysine substitutions, J. Pept. Sci. 21 (2015) 779-785. https://doi.org/10.1002/psc.2807.

Crossref Google Scholar
[33]

S.J. Walmsley, P.A. Rudnick, Y. Liang, et al., Comprehensive analysis of protein digestion using six trypsins reveals the origin of trypsin as a significant source of variability in proteomics, J. Proteome Res. 12 (2013) 5666-5680. https://doi.org/10.1021/pr400611h.

Crossref Google Scholar
[34]
L. Mora, M.C. Aristoy, F. Toldrá, Bioactive peptides in foods, in: B. Caballero, P.M. Finglas and F. Toldrá (Eds.), Encyclopedia of Food and Health, Academic Press, Oxford, 2016, pp. 395-400, https://doi.org/10.1016/B978-0-12-384947-2.00067-2.
Crossref
[35]

C. Millan-Linares Mdel, M. Yust Mdel, J.M. Alcaide-Hidalgo, et al., Lupine protein hydrolysates inhibit enzymes involved in the inflammatory pathway, Food Chem. 151 (2014) 141-147. https://doi.org/10.1016/j.foodchem.2013.11.053.

Crossref Google Scholar
[36]

J.L. Zhang, F.F. Zhou, Y.Z. Li, et al., New sesquiterpenoids with COX-2 inhibitory activity from the medical plant Physalis alkekengi L. var. franchetii, Fitoterapia 141 (2020) 104470. https://doi.org/10.1016/j.fitote.2020.104470.

Crossref Google Scholar
[37]

W. Liu, H. Li, Y. Wen, et al., Molecular mechanism for the α-glucosidase inhibitory effect of wheat germ peptides, J. Agric. Food Chem. 69 (2021) 15231-15239. https://doi.org/10.1021/acs.jafc.1c06098.

Crossref Google Scholar
[38]

G. Fricker, C. Bruns, J. Munzer, et al., Intestinal absorption of the octapeptide SMS 201–995 visualized by fluorescence derivatization, Gastroenterology 100 (1991) 1544-1552. https://doi.org/10.1016/0016-5085(91)90651-z.

Crossref Google Scholar
[39]

A.M. Ghribi, A. Sila, R. Przybylski, et al., Purification and identification of novel antioxidant peptides from enzymatic hydrolysate of chickpea (Cicer arietinum L.) protein concentrate, J. Funct. Foods 12 (2015) 516-525. https://doi.org/10.1016/j.jff.2014.12.011.

Crossref Google Scholar
[40]

H. You, T. Wu, W. Wang, et al., Preparation and identification of dipeptidyl peptidase IV inhibitory peptides from quinoa protein, Food Res. Int. 156 (2022) 111176. https://doi.org/10.1016/j.foodres.2022.111176.

Crossref Google Scholar
[41]

J. Feng, Y.L. Ma, P. Sun, et al., Purification and characterisation of α-glucosidase inhibitory peptides from defatted camellia seed cake, Int. J. Food Sci. Technol. 56 (2020) 138-147. https://doi.org/10.1111/ijfs.14613.

Crossref Google Scholar
[42]

R. Jarapula, K. Gangarapu, S. Manda, et al., Synthesis, in vivo antiinflammatory activity, and molecular docking studies of new isatin derivatives, Int. J. Med. Chem. 2016 (2016) 2181027. https://doi.org/10.1155/2016/2181027.

Crossref Google Scholar
[43]

F.A. Ragab, H.I. Heiba, M.G. El-Gazzar, et al., Anti-inflammatory, analgesic and COX-2 inhibitory activity of novel thiadiazoles in irradiated rats, J. Photochem. Photobiol. B: Biol. 166 (2016) 285-300. https://doi.org/10.1016/j.jphotobiol.2016.12.007.

Crossref Google Scholar
[44]

D.K. Yadav, Saloni, P. Sharma, et al., Studies of the benzopyran class of selective COX-2 inhibitors using 3D-QSAR and molecular docking, Arch. Pharm. Res. 41 (2018) 1178-1189. https://doi.org/10.1007/s12272-017-0945-7.

Crossref Google Scholar
[45]

T. Sumaryada, A. Astarina, L. Ambarsari, Molecular docking and physicochemical analysis of the active compounds of soursop (Annona muricata Linn) for an anti-breast cancer agent, Biointerface Res. Appl. Chem. 11 (2021) 11380-11389. https://doi.org/10.33263/BRIAC114.1138011389.

Crossref Google Scholar
[46]

M.A. Ahmaditaba, S. Shahosseini, B. Daraei, et al., Design, synthesis, and biological evaluation of new peptide analogues as selective COX-2 inhibitors, Arch. Pharm. 350 (2017) 1700158. https://doi.org/10.1002/ardp.201700158.

Crossref Google Scholar
[47]

S. Gupta, A.K. Sharma, V. Shastri, et al., Prediction of anti-inflammatory proteins/peptides: an insilico approach, J. Transl. Med. 15 (2017) 7. https://doi.org/10.1186/s12967-016-1103-6.

Crossref Google Scholar
[48]

S.R.C.K. Rajendran, B. Mason, C.C. Udenigwe, Peptidomics of peptic digest of selected potato tuber proteins: Post-translational modifications and limited cleavage specificity, J. Agric. Food Chem. 64 (2016) 2432-2437. https://doi.org/10.1021/acs.jafc.6b00418.

Crossref Google Scholar
[49]

A.B. Nongonierma, S. Paolella, P. Mudgil, et al., Identification of novel dipeptidyl peptidase IV (DPP-IV) inhibitory peptides in camel milk protein hydrolysates, Food Chem. 244 (2017) 340-348. https://doi.org/10.1016/j.foodchem.2017.10.033.

Crossref Google Scholar
[50]

M.M. Wang, Y.N. Li, W.K. Ming, et al., Bioassay-guided isolation of human carboxylesterase 2 inhibitory and antioxidant constituents from Laportea bulbifera: inhibition interactions and molecular mechanism, Arab. J. Chem. 15 (2022) 103723. https://doi.org/10.1016/j.arabjc.2022.103723.

Crossref Google Scholar
Food Science and Human Wellness
Volume 13 Issue 3,
May 2024
Pages 1636-1644
DOI: 10.26599/FSHW.2022.9250143
Cite this article:
Hong Z, Xie J, Tao L, et al. Exploration of cyclooxygenase-2 inhibitory peptides from walnut dreg proteins based on in silico and in vitro analysis. Food Science and Human Wellness, 2024, 13(3): 1636-1644. https://doi.org/10.26599/FSHW.2022.9250143
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COX-2 binding sites of the peptides determined by virtual screening.

Peptide Score (kcal/mol) Binding sites with hydrogen bonds and their length Binding sites with hydrophobic interactions
AGFP −9.1 Tyr341 (3.30, 3.30 Å), Tyr371 (3.01 Å), Ser516 (3.00 Å) Leu78, Arg106, Gly512, Ser339, His75, Trp373, Val509, Leu338, Phe504, Val335, Val102, Ala513, Leu345
FPGA −8.1 Tyr341 (3.27 Å), Arg106 (3.54, 2.81, 2.86 Å), Ser105 (2.70 Å) Ala513, Ser339, Leu338, Val509, Phe504, Val335, Leu345, Leu517, Tyr101, Val74, Leu78, Val102
LFPD −8.4 Tyr341 (3.20, 2.74, 3.19 Å), Val509 (3.37 Å) Leu78, Val74, Val102, Ser516, Leu345, Ala513, Leu517, Leu370, Tyr371, Phe367, Gly512, Val335, Trp373, Leu338, Ser339, Phe504
VGFP −9.2 Tyr371 (3.07 Å), Ser516 (3.06, 2.73 Å), His75 (3.42 Å) Ala502, Leu517, Leu345, Val102, Val335, Arg106, Tyr341, Ser339, Val509, Phe504, Leu338, Ala513, Gly512, Gln178, Trp373