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Preparation method of lactic acid bacteria fermented wheat germ extract and its antitumor effect

A lactic acid bacteria fermentation and extract technology, applied in the preparation of functional food or medicine, and the field of fermented wheat germ extract, can solve the problem that the anti-tumor extract has not been patented or reported in other literatures, etc., and achieve significant anti-tumor effect, production Low-cost, wide-ranging effects

Active Publication Date: 2015-08-05
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The Japanese patent mentioned above is an anti-tumor mixed fermentation extract obtained by fermenting various germs with Aspergillus wheat, while the Hungarian and US patents are anti-tumor fermented extracts obtained by fermenting wheat germ with baker’s yeast, and anti-tumor extracts obtained by fermenting wheat germ with lactic acid bacteria There are no patents or other literature reports yet

Method used

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  • Preparation method of lactic acid bacteria fermented wheat germ extract and its antitumor effect
  • Preparation method of lactic acid bacteria fermented wheat germ extract and its antitumor effect
  • Preparation method of lactic acid bacteria fermented wheat germ extract and its antitumor effect

Examples

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Effect test

Embodiment 1

[0042] Weigh 20.0g of crushed fresh wheat germ powder into a 500mL conical flask, add 100ml of distilled water to it, then add 0.4g of Lactobacillus plantarum Dy-1, and mix well; seal the conical flask with gauze , placed in a constant temperature incubator and fermented at 30° C. for 24 hours, centrifuged at 12,000 rpm / min for 20 minutes to collect the supernatant and freeze-dry.

[0043] Weigh 0.5g of lyophilized powder and dissolve it in 50ml of deionized water, pass through a 0.22um filter membrane, and filter the filtrate according to the following 0.125mg / mL, 0.25mg / mL, 0.5mg / mL, 1.0mg / mL, 2.0mg / ml , 4.0mg / ml were added to HT-29 colon cancer cells, and the cells were cultured for 24h, 48h, 72h, and the inhibition rate of LFWGE on colon cancer HT-29 cells was determined by MTT method ( figure 1 ). The results showed that the concentration of the extract was positively correlated with the inhibition rate of tumor cell proliferation, and the maximum inhibition rate reached...

Embodiment 2

[0046] The filtrate in Example 1 was also added to HT-29 colon cancer cells at 0.125 mg / ml, 0.25 mg / ml, 0.5 mg / ml, 1 mg / ml, 2 mg / ml, and 4 mg / ml, respectively, and the cells were cultured for 24 h, 48h, 72h, and the inhibitory rate of LFWGE to colon cancer HT-29 cells was determined by MTT method. The results showed that the concentration of the extract was positively correlated with the inhibition rate of tumor cell proliferation, and the maximum inhibition rate reached 89.6%.

[0047] 1 mg / ml LFWGE was added to SGC-7901 cells, and after culturing the cells for 48 hours, the cells were collected, and the apoptosis of the cells was detected by flow cytometry. Early apoptosis was 48.5%, and late apoptosis was 22.4%. Add 0.2mg / ml, 0.4mg / ml, 0.8mg / ml LFWGE to gastric cancer cells, culture the cells for 48 hours, collect the cells, and detect the cell cycle by flow cytometry. The results showed that with the increase of concentration, G 1 The S phase is also prolonged, while th...

Embodiment 3

[0049] 100 g of crushed fresh wheat germ was mixed with 1 L of distilled water and 4 g of Lactobacillus plantarum, and the mixture was fermented at 30° C. for 48 hours at a speed of 250 rpm. The obtained fermentation product was centrifuged at 10000 rpm for 15 min, and the supernatant was collected and freeze-dried to obtain LFWGE freeze-dried powder. The distribution of water-soluble proteins in LFWGE was detected by SDS-PAGE ( Figure 4 ). In order to separate the protein components in LFWGE, (NH4) 2 SO 4 The precipitation method precipitates the protein in the LFWGE aqueous solution, dissolves the precipitate in PBS, and passes the resulting solution through SephadexG-50 and SurpedexS-200 gel filtration chromatography to collect peak 1 and peak 2 components respectively ( Figure 5 ) and freeze-dried. The inhibitory rate of peak 1 and peak 2 components to HT-29 cells was measured by MTT method. The results showed that Peak 1 had a significant inhibitory rate on HT-29 c...

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Abstract

The invention discloses a preparation method of a lactobacillus fermented wheat germ extract and anti-tumor effects of the lactobacillus fermented wheat germ extract, and belongs to the field of food biotechnology. According to the preparation method, LactobacillusplantarumDy-1 (CGMCC No.6016) screened from pickled vegetable is used as a fermentation strain; and fermentation conditions are optimized so as to obtain the LFWGE with anti-tumor effect and relatively high yield. Experiments on a plurality of kinds of human tumor cells and tumor-bearing animals of the human tumor cells are performed by using the LFWGE, and the results show that the LFWGE possesses significant inhibition effects on tumor cell proliferation; and the composition of the LFWGE is also analyzed, and the LFWGE is rich in ingredients such as quinones, saccharides, proteins and polypeptides. According to the preparation method, wheat germ is subjected to simple pretreatment (non-high temperature heating process), production cost is low, direct adding fermentation method is employed. The preparation method is convenient, fast, safe and stable, and is convenient for quality control.

Description

technical field [0001] The invention relates to the field of food biotechnology, specifically refers to the preparation of fermented wheat germ extract (LFWGE) which can inhibit tumor cell proliferation and tumor growth by using wheat germ (wheat germ for short) as raw material to realize the biotransformation of nutrients through lactic acid bacteria fermentation , the product of the invention can be used in the preparation of functional food or medicine. Background technique [0002] At present, the number of deaths caused by malignant tumors in China accounts for 1 / 4 of the world's total, and the number is still rising. The treatment of cancer mainly includes surgery, chemotherapy, radiotherapy, and targeted therapy (immunotherapy, gene therapy, angiogenesis inhibitors, and targeted therapy), and the side effects still bring a lot of pain to patients. Therefore, it is the desire of cancer patients to develop anti-tumor active substances and functional foods that are natu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A23L1/172A23L1/30A23L33/10
Inventor 董英张家艳徐斌周兴华肖香
Owner JIANGSU UNIV
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