Bioactive peptides with apn inhibitory effect
A technology of bioactive peptides and inhibition, which is applied in the direction of peptides, drug combinations, allergic diseases, etc., can solve the problems of high price, long research cycle of separation and purification of bioactive peptides, etc., to achieve good immunity activity of the body, promote anti-inflammatory Effect of tumor function, ability to improve
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Embodiment 1
[0025] Example 1. Screening of Active Peptides with APN Inhibition
[0026] Virtual enzymolysis of egg proteins (ovomucin, avidin, ovotransferrin) by pepsin and trypsin using the online tool ExPASy PeptideCutter (http: / / web.expasy.org / peptide_cutter / ) The peptide sequences generated after the solution were compared with the known APN inhibitory peptides in the BIOPEP (http: / / www.uwm.edu.pl / biochemia / index.php / en / biopep) database, and three unreported peptides were screened. Peptide CDR, SDF, ADF, FYQ, GEF, NTF, LWK, MIR, NAF, AQF, CNR. Water solubility, biological activity and ADMET (absorption, distribution , metabolism, excretion, toxicity) properties prediction. The screened tripeptides CDR, SDF, GEF, ADF, LWK, and CNR have good solubility, activity score higher than 0.5, and non-toxicity.
[0027]The crystal structure (4FYR) of APN was obtained from the PDB database (http: / / www.rcsb.org / ), and it was used as a protein target, and the tight binding ability of the peptide...
Embodiment 2
[0028] Example 2. Identification of APN inhibitory activity in vitro
[0029] The APN inhibitory activities of peptides CNR, CNR and GEF were determined by microplate reader. Take 20 μL of active peptide, add 20 μL of a standard product with a concentration of 60 U / L and mix well, preheat in a 37°C constant temperature water bath for 10 minutes, and then take 20 μL of the mixture. Add 30 μL of sample diluent to the enzyme-labeled plate, mix well with the mixture, add 100 μL of enzyme-labeled reagent, seal the plate with a sealing film, and incubate at 37°C for 60 minutes. Dilute the concentrated washing solution 20 times with distilled water before use. Peel off the sealing film, discard the liquid, shake dry, fill each well with washing solution, let it stand for 30 seconds and then discard, repeat this 5 times, and pat dry. First add 50 μL of chromogenic reagent A to each well, then add 50 μL of chromogenic reagent B, shake and mix well, and develop color at 37°C in the da...
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