Application of tilapia protein peptide to preparation of immune preparation
A technology of immune preparations and tilapia, which is applied in the direction of hydrolyzed protein components, peptide/protein components, medical preparations containing active ingredients, etc., and can solve problems such as environmental pollution and waste of resources
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Embodiment 1
[0046] The preparation of the protein peptide of embodiment 1 tilapia
[0047] Method for enzymatically hydrolyzing tilapia protein peptides from tilapia leftovers:
[0048] (1) Put the leftovers of tilapia into boiling water and cook for 10 minutes, take it out and cool, and obtain the tilapia material.
[0049] (2) Pulverize the cooled tilapia material, wash and stir with a 40-mesh mesh frame, and repeat 3-4 times.
[0050] (3) Mix according to the ratio of the mass of crushed tilapia material to the volume of water of 1:5, use a colloid mill to refine the pulp, adjust the system temperature to 45°C-60°C, pH 7.0-9.0, add trypsin Mass concentration 0.1-0.3%, stirring enzymolysis for 1h, then adding flavor protease mass concentration 0.1-0.3%, stirring enzymolysis for 3-6h, heating at 80°C-95°C for 10-25min, centrifuging to get supernatant.
[0051] (4) Concentrate the supernatant by nanofiltration, decolorize the obtained nanofiltration concentrate with activated carbon, evap...
Embodiment 2
[0055] In this experiment, the immunopromoting activity of tilapia protein peptide was verified.
[0056] 1. Experimental raw materials and related reagents
[0057] The tilapia protein peptides used in this experiment are from Shanwei Weiming Biotechnology Co., Ltd. (batch number 20180508-1). The preparation method is as shown in Example 1. The content of oligopeptides: more than 95.1%, detected and analyzed by ESI-LCMS mass spectrometry Its molecular weight range: 470-1417Da (such as diagram 2-1 with 2-2 shown); high-glucose DMEM medium (article number SH30022.01), penicillin, and streptomycin were purchased from Hyclone Company of the United States; fetal bovine serum was purchased from BI Company; NO detection kit, CCK-8 kit, and cell lysate were purchased from The iNOS monoclonal antibody and goat rabbit secondary antibody were purchased from CST Company in Germany; the ECL luminescent liquid was purchased from Thermo Fisher Scientific Company; LPS and other commonly u...
Embodiment 3
[0085] Except for the batch of tilapia protein peptide purchased from Shanwei Weiming Biotechnology Co., Ltd. (the content of oligopeptide: more than 95.1%, and the molecular weight range of ESI-LCMS mass spectrometry: 470-1417Da), the rest were Same as Example 1. NO release see Figure 5 .
[0086] Example 3
[0087] Except for the batch of tilapia protein peptide purchased from Shanwei Weiming Biotechnology Co., Ltd. (the content of oligopeptide: more than 95.1%, and the molecular weight range of ESI-LCMS mass spectrometry: 470-1417Da), the rest were Same as Example 1. NO release see Image 6 .
[0088] Such as Image 6 with image 3As shown, the present invention uses different batches of commercially available tilapia protein peptides, all of which have the effect of increasing NO release, which means that the tilapia protein peptide has a good immune regulation effect.
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