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Open Access

Virtual screening and directional preparation of xanthine oxidase inhibitory peptides derived from hemp seed protein

Xiaoni Huang1, Jiajun Liao1, Ping Shi, Xiaodong Pei, Chenghua Wang()
College of Light Industry and Food Engineering, Guangxi University, Nanning 530004, China

1 These authors contributed equally to this work.

Peer review under responsibility of Tsinghua University Press.

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Abstract

The traditional nutritional and medical hemp (Cannabis sativa L.) seed protein were explored for the discovery and directional preparation of new xanthine oxidase inhibitory (XOI) peptides by structure-based virtual screening, compound synthesis, in vitro bioassay and proteolysis. Six subtypes of hemp seed edestin and albumin were in silico hydrolyzed by 29 proteases, and 192 encrypted bioactive peptides were screened out. Six peptides showed to be XOI peptides, of which four (about 67%) were released by elastase hydrolysis. The peptide DDNPRRFY displayed the highest XOI activity (IC50 = (2.10 ± 0.06) mg/mL), acting as a mixed inhibitor. The pancreatic elastase directionally prepared XOI hemp seed protein hydrolysates, from which 6 high-abundance XOI peptides encrypted 3 virtually-screened ones including the DDNPRRFY. The novel outstanding hemp seed protein-derived XOI peptides and their virtual screening and directed preparation methods provide a promising and applicable approach to conveniently and efficiently explore food-derived bioactive peptides.

Keywords

Hemp seed proteinAnti-goutBioactive peptideVirtual screeningProteolysis

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Food Science and Human Wellness
Volume 13 Issue 6,
November 2024
Pages 3652-3660
DOI: 10.26599/FSHW.2023.9250047
Cite this article:
Huang X, Liao J, Shi P, et al. Virtual screening and directional preparation of xanthine oxidase inhibitory peptides derived from hemp seed protein. Food Science and Human Wellness, 2024, 13(6): 3652-3660. https://doi.org/10.26599/FSHW.2023.9250047
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Interaction between peptide inhibitor and XOD and molecular docking score results.

Peptide sequence Hydrogen bond interaction Hydrophobic interaction π-π stacking interaction Affinity score Corresponding protease
Related amino acids Bond length (Å) Related amino acids Related amino acids
Notes: “-” indicates there is no such type of interaction between peptide inhibitor and XOD.
PRRFYI Glu1143, Asp872, Met770, Glu802, Leu873 3.28, 3.40, 4.27, 3.20, 3.23 Leu1014 – –8.64 Elastase protease
Leu648
YWCNNDG Glu802, Leu873, Glu879, Ser876 2.85, 3.38, 2.66, 3.00 Lys771 Phe914 –9.22 Bromelain
Leu1014
DDNPRRFY Leu1138, Glu879, Thr1010, Lys771, Glu802, Arg1134, Leu873, Ser1141 2.86, 3.33, 2.94, 2.82, 2.47, 2.94, 3.47 Leu1014 Phe914 –9.44 Elastase protease
FCFFL Glu802, Glu879, Ser876 3.00, 2.67, 2.60 Phe649 Phe914 –9.07 Elastase protease
EGFEWVSFK Glu802, Glu879, Lys771 2.73, 3.08, 2.52 Phe1013 Phe914 –9.37 Trypsin
QSCHFRTP Glu802, Glu879, Arg880, His875, Val1011, Ser876, Thr1010 3.28, 2.75, 3.48 Phe649Leu1014 – –7.19 Prolyl oligopeptidase
SADDEDNYYYGG Glu1143, Glu802 3.12, 2.78 – – –7.94 Prolyl oligopeptidase
PYY Ser876, Phe1009, Val1011, Glu802 2.55, 3.00, 3.02 Leu1014 Phe914 –7.33 Elastase protease

Inhibitory kinetic parameters of DDNPRRFY peptide.

Polypeptideinhibitor concentration (mg/mL) Apparent Vmax(min–1) Apparent Km (mmol/L) Doublereciprocal curve equation
0.0 0.043 ± 0.002 0.090 ± 0.012 y =0.59x + 31.00, R2=0.98
0.8 0.030 ± 0.001 0.058 ± 0.010 y =1.57x + 35.62, R2=0.95
1.6 0.028 ± 0.001 0.068 ± 0.013 y =2.22x + 36.43, R2=0.95
2.4 0.026 ± 0.002 0.093 ± 0.012 y =3.56x + 43.93, R2=0.99

In silico evaluation of bioactive peptides screened from directionally prepared HPHs by pancreatic elastase.

Peptide sequence ADMET properties Water solubility Toxicity Carcinogenicity Coverage in hemp seed polypeptide library (%)
Note: B indicates the blood-brain barrier penetration ability, H indicates the absorption ability of small intestine, in which the expression with “+” is better, and the expression with “–” is weak. G indicates good water solubility, while P indicates poor, and Non indicates no toxicity or carcinogenicity.
INNQNQLDMSPRRFYI H+/B– G Non Non 3.6
KMAEKEGFEWVSFKTNDRA H+/B– G Non Non 14.9
RENIGDPSRADV H+/B– G Non Non 2.8
RQPNRRVECEAGV H+/B– G Non Non 1.6
QQNGLHLPSYTNTPQL H+/B– P Non Non 4.8
RQGQⅣTVPQNHA H+/B– P Non Non 2.4