Two kinds of ACE inhibitory peptides derived from Larimichthys crocea connetin

A technology for inhibiting peptides and large yellow croakers, applied in peptide/protein ingredients, protein-containing food ingredients, medical preparations containing active ingredients, etc., can solve the problems of long time-consuming and high cost, and achieve reduced quantity and high adjuvant treatment Blood pressure, the effect of increasing the probability of successful screening

Inactive Publication Date: 2018-05-15
BOHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But its disadvantage is that it takes a long time and hi...

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  • Two kinds of ACE inhibitory peptides derived from Larimichthys crocea connetin
  • Two kinds of ACE inhibitory peptides derived from Larimichthys crocea connetin
  • Two kinds of ACE inhibitory peptides derived from Larimichthys crocea connetin

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Example 1 Selection of raw materials

[0024] Search for protein sequences related to large yellow croaker from the NCBI database for analysis, and the selection is based on the availability of sequence information of the parent protein and an important component of the large yellow croaker muscle protein. The final selection of the large yellow croaker Titin amino acid (ACCESSION: KKF25250), the amino acid number is 17943.

Embodiment 2 3

[0025] Example 2 Screening and activity determination of tripeptides

[0026] Pepsin (pH 1.3) and trypsin were used to virtually hydrolyze the troponin by ExPASy PeptideCutter, all tripeptides were selected, and the activity score was performed in PeptideRanker, and the 50 tripeptides with high scores were selected.

[0027] Use online tools Innovagen, ToxinPred and admetSAR to predict water solubility, toxicity and ADMET properties, respectively. A non-toxic, good water-soluble tripeptide was selected. In ADMET prediction, it focuses on the determination of good intestinal absorption (HIA+) and the tripeptide (could penetrate the Blood-Brain Barrier, BBB+) that can penetrate the Blood-Brain Barrier (BBB+) as potential high-efficiency ACE inhibitor peptide candidates. Finally selected 8 required tripeptide sequences, docked with ACE (PDB ID: 1A1B) in SwissDock, and selected the two tripeptides WAR and WQR with the lowest docking energy value, with ΔG of -10.40 and -9.57kcal, respe...

Embodiment 3

[0033] Example 3 Verification of in vitro activity

[0034] The ACE inhibitory activity of WAR and WQR was verified by high performance liquid chromatography. Take the hippuryl histidine leucine (HHL) substrate solution, add the inhibitor and mix well, preheat it in a constant temperature water bath at 37°C for 3 to 5 minutes, then add the ACE solution to mix well, keep it at 37°C for 30 minutes, then add The reaction was terminated with 1mol / L HCl to obtain a reaction solution. At the same time, boric acid buffer was used instead of inhibitor solution as a blank control group. The reaction solution was directly analyzed by the HPLC system.

[0035] Chromatographic conditions: column temperature 25°C, flow rate 0.5mL / min, mobile phase acetonitrile: water 25:75 isocratic elution, detection wavelength 228nm.

[0036] The in vitro ACE inhibitory activities of WAR and WQR were verified through experiments, which were 31.17±0.79 and 231.33±0.02μM, respectively.

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Abstract

The invention mainly relates to Larimichthys crocea connetin source ACE inhibitory peptide sequences and applications thereof. The two kinds of peptide sequences respectively are WAR and WQR, and establish a basis for prevention and treatment of hypertension. Different on-line databases are used, and after multiple times of screening, the inhibitory peptides with high efficient ACE inhibition activity are obtained. WAR and WQR are artificially synthesized, in order to verify external ACE inhibition activity of the peptides by using a high performance liquid chromatography method. The inventionrelates to two kinds of active peptides with good external ACE inhibition activity. The two kinds of peptide sequences are used for developing health food with blood pressure adjusting effects, and the purpose for prevention, control, alleviation and auxiliary treatment of hypertension is realized by long-term usage.

Description

Technical field [0001] The present invention belongs to the field of biologically active peptides, and mainly relates to two ACE inhibitory peptides derived from large yellow croaker Titin and the application of the peptides. Background technique [0002] Hypertension is a symptom of chronic arterial hypertension and causes heart failure, vision loss, and heart palpitations. In the human body, angiotensin-I converting enzyme (ACE) converts the inactive ACE-I into the vasoconstrictor ACE-II with pressor activity, and inactivates vasodilator kinin, leading to arterial Contraction, blood pressure rises. Therefore, inhibiting the activity of ACE is one of the methods to treat hypertension. Chemically synthesized ACE inhibitors such as captopril, lisinopril, enalapril and fosinopril are drugs used to treat hypertension. However, these drugs can cause certain side effects, including cough, rash, and angioedema, and ACE inhibitory peptides derived from food proteins can improve this ...

Claims

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Application Information

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IPC IPC(8): C07K5/097A61K38/06A61P9/12A23L33/18
CPCA23V2200/00A61K38/00A23L33/18C07K5/0821A23V2200/326A23V2250/55
Inventor 于志鹏樊玥赵文竹
Owner BOHAI UNIV
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