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Preparation method of small water turtle anti-tumor polypeptide

An anti-tumor and money technology, which is applied in the field of preparation of anti-tumor polypeptides of the tortoise, can solve the problems of large toxic and side effects and unsatisfactory treatment effect.

Active Publication Date: 2015-05-20
广东省几何细胞生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Due to the continuous increase in the incidence of malignant tumors in recent years, and the traditional chemotherapeutic drugs not only have large toxic and side effects but also have unsatisfactory therapeutic effects, bioactive peptides have been recognized at home and abroad for their advantages of high activity, small side effects, and strong pertinence. Research and concern of scientific researchers

Method used

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  • Preparation method of small water turtle anti-tumor polypeptide
  • Preparation method of small water turtle anti-tumor polypeptide
  • Preparation method of small water turtle anti-tumor polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) Take 5 g of golden turtle meat and ultrapure water, mix and stir to prepare a mixed solution with a concentration of 10% (w / v), add trypsin for enzymatic hydrolysis, the hydrolysis conditions are: PH=7, enzymatic hydrolysis temperature 35℃, enzymatic hydrolysis time is 5 hours, and the concentration ratio of enzyme to substrate is 0.2% (w / w). In this process, 0.05 mol / L NaOH and HCl are used to adjust the pH value of the reaction system, and control the pH value between pH±0.05. After the enzymolysis is completed, the enzyme is inactivated in 80℃ water for 10 minutes, cooled to room temperature, and the enzymatic hydrolysis The solution was centrifuged at 7000 r / min for 30 min, and the supernatant was taken.

[0027] (2) Take the enzymatic hydrolysate obtained in step (1) and use the ultrafiltration membranes with molecular weight cutoffs of 10 KD, 5 KD and 3 KD to filter (CO 2 The pressure is 0.1 Mpa at room temperature), so as to obtain the enzymatic hydrolysate of P...

Embodiment 2

[0031] (1) Take 10 g of golden turtle meat and ultrapure water, mix and stir to prepare a mixed solution with a concentration of 15% (w / v), add trypsin for enzymatic hydrolysis, the hydrolysis conditions are: PH=8, enzymatic hydrolysis temperature 30℃, enzymolysis time is 10 h, the concentration ratio of enzyme and substrate is 0.1% (w / w). During this process, 0.1 mol / L NaOH and HCl were used to adjust the pH value of the reaction system, and the pH value was controlled between pH ± 0.05. After enzymolysis, the enzyme was inactivated in 95℃ water for 15 min, cooled to room temperature, and the enzymatic hydrolysis The solution was centrifuged at 6000 r / min for 35 min, and the supernatant was taken.

[0032] (2) Take the enzymatic hydrolysate obtained in step (1) and use the ultrafiltration membranes with molecular weight cutoffs of 10 KD, 5 KD and 3 KD to filter (CO 2 The pressure is 0.22 Mpa at room temperature), so as to obtain the enzymatic hydrolysate of P. vulgaris polypepti...

Embodiment 3

[0035] (1) Mix 18 g of golden turtle meat and ultrapure water and stir to prepare a mixed solution with a concentration of 30% (w / v). Add trypsin for enzymatic hydrolysis. The hydrolysis conditions are: PH=9, enzymatic hydrolysis temperature 60°C, enzymolysis time 15 h, concentration ratio of enzyme to substrate is 0.5% (w / w). During the process, 0.2 mol / L NaOH and HCl were used to adjust the pH value of the reaction system, and the pH value was controlled between pH ± 0.05. After enzymolysis, the enzyme was inactivated in water at 100°C for 20 min, cooled to room temperature, and the enzymatic hydrolysis The solution was centrifuged at 8000 r / min for 60 min, and the supernatant was taken.

[0036] (2) Take the enzymatic hydrolysate obtained in step (1) and use the ultrafiltration membranes with molecular weight cutoffs of 10 KD, 5 KD and 3 KD to filter (CO 2 The pressure is 0.25 Mpa at room temperature), so as to obtain the enzymatic hydrolysate of the golden tortoise polypeptid...

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Abstract

The invention provides a preparation method of a small water turtle anti-tumor polypeptide. The method comprises the following steps: mixing 3 to 20g of meat paste of small water turtle and ultrapure water; agitating to obtain a mixed solution at concentration of 10 to 50% (w / v); adding trypsin to perform enzymolysis; deactivating enzyme after enzymolysis; cooling until reaching room temperature; centrifuging enzymatic hydrolysate; collecting supernate; sequentially filtering through ultrafiltration membranes which respectively have molecular cut off of 10KD, 5KD and 3KD, so as to obtain three enzymatic hydrolysates; performing sephadex G-15 column chromatography for 0-3K enzymatic hydrolysate; eluting with water; collecting four polypeptide components under a certain detection wavelength so as to obtain small water turtle anti-tumor polypeptide. The prepared small water turtle anti-tumor polypeptide is beneficial for the development and utilization of anti-tumor drugs and health foods.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method of a money turtle anti-tumor polypeptide. Background technique [0002] The money turtle (Cuora Trifasciata), also known as the three-lined box turtle, has extremely high medicinal value. In ancient Eastern Han Dynasty "Shen Nong's Materia Medica" and "Shan Hai Jing" in the Warring States Period, there are records of the edible and medicinal use of turtles; the golden turtle has obvious effects on detoxification, anti-inflammatory, liver and kidney, and is often used for health care and beauty, prevention and treatment of tumors and longevity. One of the main medicines for fitness. The active factor contained in the money turtle is relatively high, which has a certain effect on the treatment of thrombocytopenia and cell death, and can inhibit the growth of cancer cells and promote blood circulation in the human body. [0003] Biologically active pep...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K1/34A61K38/01A23L1/29A61P35/00A23L33/00
Inventor 张学武何胜洁
Owner 广东省几何细胞生物科技有限公司
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