A kind of active peptide with blood lipid-lowering function derived from Chinese wood beetle and its preparation method and application
A technology of Jitu soft-shelled turtle and active peptides, applied in the field of active peptides, can solve the problems of allergies and low protein absorption efficiency, and achieve the effects of increasing absorption efficiency, high purity, and eliminating immunogenic reactions.
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Embodiment 1
[0037] Embodiment 1 preparation of active peptide
[0038] (1) Treatment of ground beetle medicinal materials
[0039] Add 10 times the weight of deionized water or distilled water to the fresh body of the wood beetle, and then shear it for 15 minutes at a shear rate of 5000 rpm and at room temperature to prepare a wood beetle slurry.
[0040] (2) Protease hydrolysis
[0041] Heat the ground beetle slurry to 100°C to sterilize for 15 minutes, then let it cool to 37°C, use 0.1mol / L HCl to adjust the pH of the solution to 2.5, add 1.5% by weight of pepsin and set it in a constant temperature enzymolysis instrument for enzymolysis, the enzymolysis time 60min, and the enzymatic hydrolysis temperature is 37°C. Then use 2.0% sodium hydroxide solution to adjust the pH to 8.5, add 1.5% by weight of trypsin and place it in a constant temperature enzymolysis instrument for enzymolysis, the enzymolysis time is 180min, and the enzymolysis temperature is 37°C to obtain the bionic ground ...
Embodiment 2
[0052] Based on the amino acid sequences of the LL-8 and LM-9 active peptides identified in Example 1, experimental samples were obtained by solid-phase synthesis for verification of blood lipid-lowering activity. Solid-phase synthetic peptide company: Shanghai Gil Biotechnology Co., Ltd.
[0053] 80 SD rats, male, were adaptively fed for 7 days, and then divided into nine groups, namely blank group, model group, Xueshuantong capsule group, ground beetle medicinal material group, LL-8 high-dose group (0.5g / kg), LL-8 low-dose group (0.25g / kg), LM-9 high-dose group (0.5g / kg), LM-9 low-dose group (0.25g / kg), except the blank group, every mouse in other groups Quantitatively administer 50 g of high-fat feed, and at the same time remove the rats in the blank group, give each rat 4.0 mL of high-fat emulsion at 9:00 am every day, feeding method of high-fat emulsion: gavage orally, rats in the blank group continue to be given ordinary feed, continuous For 45 days, after giving high-f...
Embodiment 3
[0059] Based on the amino acid sequence of the LL-8 active peptide identified in Example 1, experimental samples were obtained by solid-phase synthesis for safety experimental verification. Solid-phase synthetic peptide company: Shanghai Gil Biotechnology Co., Ltd.
[0060] Healthy SD rats, 250-300g, 40, were adaptively fed for 7 days, and then set up as blank group, normal saline group, LL-8 group, and LL-8 group were given a large dose of 20.0g / 200g of drugs, administration Method: subcutaneous injection; the normal saline group was also given the same volume of normal saline by subcutaneous injection for 12 days. body weight, body temperature. At the same time, the weight of immune organs such as thymus and spleen of rats in each group was measured to measure the safety of LL-8.
[0061] Table 2 Short-term safety evaluation results of body temperature changes in rats in each group
[0062]
[0063]
[0064] Table 3 Organ weight results of rats in each group
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