Immunoactive peptide of saiteng fruit and preparation method thereof
A technology of immunoactive peptides and fenugreek fruit, applied in the field of immunoactive peptides, can solve the problems of limited development and utilization of protein resources, and achieve the effect of strong function and high purity
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Embodiment 1
[0057] The selection of the optimal process condition of embodiment 1 Uteng fruit albumin
[0058] Pulverize satoshi fruit meal, take satoshi fruit meal powder and place it in a 24L extraction kettle of supercritical extraction equipment, extract for 120min under the conditions of 20MPa and 32°C, CO 2 The flow rate was 60 mL / min. Discard the extracted sachaetra fruit oil, take out the sachaeto fruit cake powder and add a certain amount of first-grade water, ultrasonically treat for 15 minutes, stir and extract at a constant temperature for a certain period of time, and centrifuge at 4000 rpm for 10 minutes to separate the supernatant , the supernatant was placed in distilled water and dialyzed at 4°C for 72 hours, then evaporated in a rotary evaporator for a certain period of time, and then vacuum freeze-dried to obtain lyophilized albumin. According to the simulation experiment, considering the interaction of factors such as solid-liquid ratio, extraction temperature, and ex...
Embodiment 2
[0065] Embodiment 2 Screening of satobumin proteolytic enzymes and selection of optimal hydrolysis conditions
[0066] The cane albumin was prepared according to the ratio of material to liquid of 1:15, and after adjusting the optimum temperature and pH, neutral protease, papain, compound protease, trypsin, gastric Protease and alkaline protease, enzymatic hydrolysis for 3 hours, and the enzyme types were screened with the degree of hydrolysis as an index. According to the simulation experiment, considering the interaction of factors such as enzymatic hydrolysis time, enzymatic hydrolysis temperature, enzyme amount and pH, it is planned to use orthogonal experiments to determine the optimal enzymatic hydrolysis conditions of albumin. In this experiment, L 9 (3 4 ) orthogonal test to complete, the program is shown in Table 5.
[0067] Experimental results such as figure 2 As shown, there is a significant difference in the degree of hydrolysis of the proteolytic hydrolyzate ...
Embodiment 3
[0070] Example 3 Isolation, Purification, Identification and Immunological Activity Evaluation of Immunoactive Peptide
[0071] 1. Ultrafiltration separation
[0072] In order to purify and enrich the active components, the hydrolyzate of saponin fruit albumen is subjected to ultrafiltration to obtain a mixed solution of polypeptides with different molecular weights. Put the enzymatic hydrolysis solution in the ultrafiltration cup, and perform ultrafiltration treatment with 10kDa and 3kDa ultrafiltration membrane modules successively. After closing the ultrafiltration cup, pressurize with nitrogen to keep the pressure at 0.2MPa. Place the obtained peptide solution on ice to ensure its activity. Subsequently, the mouse macrophage model RAW264.7 was used to evaluate the in vitro immune activity of each ultrafiltration fraction at a concentration of 100 μg / mL, and compared with the blank control group, 1 μg / mL lipopolysaccharide (LPS) and the same concentration without ultrafilt...
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