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Bitter gourd polypeptide for inhibiting tumor cell proliferation as well as preparation method and application thereof

A technology of tumor cell proliferation and bitter gourd polypeptide, applied in the biological field, can solve the problems of poor control of peptides, uncontrollable safety, severe reaction conditions, etc., achieve anti-tumor biological activity and strong effect, no toxic side effects, The effect of high biological potency

Active Publication Date: 2018-10-26
苏秀兰
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a preparation method of bitter gourd polypeptide that inhibits the proliferation of tumor cells, to solve the problem that the existing preparation method has severe reaction conditions, the obtained peptide cannot be well controlled, the hydrolysis time is long or the safety is uncontrollable The problem

Method used

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  • Bitter gourd polypeptide for inhibiting tumor cell proliferation as well as preparation method and application thereof
  • Bitter gourd polypeptide for inhibiting tumor cell proliferation as well as preparation method and application thereof
  • Bitter gourd polypeptide for inhibiting tumor cell proliferation as well as preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] This embodiment provides a method for preparing a bitter gourd polypeptide that inhibits tumor cell proliferation, such as figure 1 As shown, the method includes the following steps:

[0044] Step 1: Wash the fresh bitter gourd with clean water, remove the stalks, and cut into small pieces for later use.

[0045] Step 2: Weigh, add the material liquid into RETSCH GM300 knife grinder at a ratio of 1:2, 2000rpm, 3min, to obtain a coarse grinding material liquid (Ultrapure water pH=7).

[0046] Step 3: Freeze the coarse ground feed liquid at -60°C for 12 hours.

[0047] Step 4: Thaw the coarse grinding material liquid, GM300 grinder, 2000rpm, 3min, filter with a 400-mesh sieve to obtain a fine grinding material liquid.

[0048] Step 5: Finely grind the material liquid, centrifuge at 4000 rpm for 15 minutes in a refrigerated centrifuge, and keep the supernatant.

[0049] Step 6: The supernatant was centrifuged in a high-speed refrigerated centrifuge at 9000 rpm for 15 mi...

Embodiment 2

[0054] This embodiment provides a method for preparing a bitter gourd polypeptide that inhibits the proliferation of tumor cells, the method comprising the following steps:

[0055] Step 1: Wash the fresh bitter gourd with clean water, remove the stalks, and cut into small pieces for later use.

[0056] Step 2: Weigh, add the material liquid into RETSCH GM300 knife grinder at a ratio of 1:3, 2000rpm, 2min, to obtain a coarse grinding material liquid (Ultrapure water pH=7).

[0057] Step 3: Freeze the coarse ground feed liquid at -40°C for 8 hours.

[0058] Step 4: Thaw the coarse grinding material liquid, GM300 grinder, 2000rpm, 2min, filter with 300 mesh screen to obtain fine grinding material liquid.

[0059] Step 5: Finely grind the material liquid, centrifuge at 4000 rpm for 10 minutes in a refrigerated centrifuge, and keep the supernatant.

[0060] Step 6: The supernatant was centrifuged in a high-speed refrigerated centrifuge at 9000 rpm for 10 minutes, and the superna...

Embodiment 3

[0065] This embodiment provides a method for preparing a bitter gourd polypeptide that inhibits the proliferation of tumor cells, the method comprising the following steps:

[0066] Step 1: Wash the fresh bitter gourd with clean water, remove the stalks, and cut into small pieces for later use.

[0067] Step 2: Weigh, add the feed liquid into RETSCH GM300 knife grinder at a ratio of 1:1, 2000rpm, 5min, to obtain a coarse grinding feed liquid (Ultrapure water pH=7).

[0068] Step 3: Freeze the coarse ground material liquid at -80°C for 24 hours.

[0069] Step 4: Thaw the coarse grinding material liquid, GM300 grinder, 2000rpm, 5min, filter with a 500-mesh sieve to obtain a fine grinding material liquid.

[0070] Step 5: Finely grind the material liquid, centrifuge at 5000 rpm for 20 minutes in a refrigerated centrifuge, and keep the supernatant.

[0071] Step 6: The supernatant was centrifuged at 10,000 rpm for 20 minutes in a high-speed refrigerated centrifuge, and the super...

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Abstract

The invention discloses bitter gourd polypeptide for inhibiting tumor cell proliferation as well as a preparation method and application thereof. The preparation method comprises the following steps:step 1: chopping bitter gourds for later use; step 2: preparing a rough grinding material solution; step 3: freezing the rough grinding material solution at -40 to -80 DEG C; step 4: unfreezing; grinding and filtering through a screen mesh to prepare a fine grinding material solution; step 5: centrifuging in a freezing centrifugal machine and taking supernatant; step 6: centrifuging in a high-speed freezing centrifugal machine and taking supernatant; step 7: carrying out ultra-filtration, wherein the film covering diameter is 10 KD and filtrate with the diameter smaller than 10 KD is the bitter gourd polypeptide. According to the method provided by the invention, cell walls can be damaged to a certain extent and the dissolution rate of active substances is high; the activity of bitter gourd bioactive peptide obtained by combining an ultra-filtration technology is relatively high and a technology is easily industrialized. Meanwhile, the raw material bitter gourd has the advantages of safety for eating, no toxic side effect, high nutrient value and the like. Furthermore, the prepared bitter gourd polypeptide has the effect of inhibiting the tumor cell proliferation.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a bitter gourd polypeptide for inhibiting tumor cell proliferation, a preparation method and application thereof. Background technique [0002] Bitter gourd is a traditional Chinese herbal medicine, which has the functions of clearing away heat and relieving heat, nourishing blood and Qi, nourishing kidney and spleen, nourishing liver and improving eyesight. For the fruit of Cucurbitaceae bitter gourd. Traditionally used in folk medicine to treat peptic ulcer, kidney stones, eczema, type 2 diabetes, and secondly, bitter melon has a high vitamin C content, which can prevent scurvy, protect cell membranes, prevent atherosclerosis, and improve the body's stress ability , protect the heart, and the active ingredients in bitter melon can inhibit the canceration of normal cells and promote the restoration of anti-cancer properties of mutant cells. Since ancient times, China has used bitt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/14C07K1/34A61P35/00A23L33/185
CPCA23V2002/00A23L33/185A61P35/00C07K1/145C07K1/34A23V2200/308
Inventor 苏秀兰韩汶延温宇旗张志惠
Owner 苏秀兰
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