Active peptide derived from myosin and its application
A myosin and active peptide technology, applied in the field of protein active peptides, can solve the problems of limited types of umami peptides and different contribution levels, and achieve the effects of high umami intensity, easy absorption and high purity
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Embodiment 1
[0029] Enzymolysis screening of embodiment 1 scallop myosin
[0030] Pepsin (EC 3.4.23.1), trypsin (EC 3.4.21.4) and chymotrypsin (EC 3.4.21.1) were selected to use on muscle by ExPASy PeptideCutter (http: / / web.expasy.org / peptide_cutter / ) The globulin sequence (GenBank: BAB40711.1) was digested to obtain 747 peptide sequences.
[0031] The toxicity of the peptides was analyzed using ToxinPred (http: / / crdd.osdd.net / raghava / / toxinpred / ). The water solubility of the peptides was analyzed by peptide property calculator (http: / / www.innovagen.com / ), and 322 non-toxic active peptides with good water solubility were screened. The taste of the umami peptide is related to the composition of amino acids, the spatial arrangement of amino acids and the molecular weight of the peptide. It contains glutamic acid and aspartic acid, and contains certain hydrophilic amino acids, and the length of the peptide chain skeleton is 2-6. , has good taste effect, and the C-terminal of the dipeptide i...
Embodiment 2
[0036] Embodiment 2 Electronic Tongue Detection
[0037] This embodiment selects monosodium glutamate (MSG) as positive control. Peptides DK, EEK, EDQK, SEGGR, QDSIGS and MSG were formulated as 0.1 mg / ml solutions. Five test sensors, AAE, CT0, CA0, C00, and AE1, and two reference sensors were selected to detect the samples. The taste of the samples was analyzed by interpolation method, and the data processing results are shown in Table 2.
[0038] Table 2 Umami and Richness values of different peptides based on electronic tongue
[0039]
[0040] The results showed that the active peptides DK, EEK, EDQK, SEGGR, and QDSIGS had better freshness effects than monosodium glutamate (MSG). Among them, peptide EDQK had the highest umami intensity. Finally, the present invention provides five umami peptides.
Embodiment 3
[0041] The chemical synthesis of embodiment 3 active peptide DK, EEK, EDQK, SEGGR, QDSIGS
[0042]In this example, peptides DK, EEK, EDQK, SEGGR, and QDSIGS were synthesized in solid phase by FMOC method. At a temperature of 22°C to 28°C and normal pressure, add Fmoc-Tyr(tBu)-Wang Resin to the polypeptide solid-phase synthesis reactor, add 10mL 25% piperidine / DMF solution to remove the amino protecting group Fmoc, and then The peptide chain is extended by reacting with the activated hydroxyl group of the next amino acid with a 2-fold excess. During the condensation process, ninhydrin is used to monitor the reaction process qualitatively by color development until the target peptide required for synthesis is obtained. Select appropriate cleavage conditions to cleave the target polypeptide from the resin and remove the side chain protecting group. After the solution is rotary evaporated, anhydrous ether is added to precipitate the polypeptide. After centrifugation, washing and v...
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