Saccharomyces polypeptide derived from saccharomyces cerevisiae var. boulardii, preparation method for saccharomyces polypeptide and application of saccharomyces polypeptide

A technology of Brady's yeast and yeast polypeptide, which is applied in the field of biomedicine and can solve problems such as little research

Active Publication Date: 2021-05-18
杭州吾尾科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is little research on Brady's polypeptide

Method used

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  • Saccharomyces polypeptide derived from saccharomyces cerevisiae var. boulardii, preparation method for saccharomyces polypeptide and application of saccharomyces polypeptide
  • Saccharomyces polypeptide derived from saccharomyces cerevisiae var. boulardii, preparation method for saccharomyces polypeptide and application of saccharomyces polypeptide
  • Saccharomyces polypeptide derived from saccharomyces cerevisiae var. boulardii, preparation method for saccharomyces polypeptide and application of saccharomyces polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1. Preparation of yeast fermentation product and extraction and identification of polypeptide FTP-21

[0020] Brady's yeast CNCM I-1079 was selected as the fermentation microorganism, corn and pomegranate peel extracts were selected as the fermentation substrate, and the fermentation parameters were adjusted as follows: temperature 25°C, humidity 75%, and pH value of neutral to acidic fermentation tank for 24 -48 hours. The detection results of the fermentation products are as follows in Table 1:

[0021]

[0022] Yeast powder was obtained by freeze-drying in a vacuum freeze-drying machine. The yeast cells are crushed by mechanical grinding, followed by heat treatment and ethanol precipitation to extract yeast protein substances. After collecting the yeast samples, use anion exchange to purify the polypeptide fermented by yeast powder. The specific steps are as follows:

[0023] Pack the column, add 10 volumes of low-salt buffer until the UV absorption val...

Embodiment 2

[0026] Example 2. Antibacterial test (MIC) of yeast polypeptide FTP-21

[0027] Bacteria (Escherichia coli E. coli K 12, Escherichia coli E. coli K 88, Salmonella Salmonella , Staphylococcus aureus S. aureus , Clostridium perfringens C. perfringens , Bifidobacterium Bifidobacterium longum and Lactobacillus acidophilus Lactobacillus acidophilus ) was inoculated on MH broth solid medium, cultured until a single colony grew, picked a single colony and inoculated in 3mL MH broth medium, put it into a shaker and cultured until the bacterial solution was turbid, and transferred 30 μL of the bacterial suspension to In 3mL of fresh MH broth medium, shake culture at constant temperature until OD 600 =0.5 or so; take 10 μL of the bacterial suspension and transfer it to 10 mL of fresh MH broth medium, vortex and mix well, and control the concentration at 1×10 5 —5×10 5 CFU / mL, used for the determination of the minimum inhibitory concentration (MIC) and TEM; the antibact...

Embodiment 3

[0038] Example 3, Cytotoxicity of Yeast Polypeptide FTP-21

[0039] Dilute the isolated and purified peripheral blood mononuclear cells (PBMCs) to 1×10 with medium RMPI1640 6 cells / mL, inoculate a 96-well plate at 90 uL per well, and set up background control (100 μL RMPI1640 medium), low-level control wells (90 μL cell suspension + 10 μL PBS) and high-level control wells (90 μL cell suspension + 10 μL 20 % TritonX-100). Incubate the 96-well plate at 37°C, 5% CO 2 Incubate in the incubator for 2 hours; add 10uL of FTP-21 and LL-37 of the corresponding concentration, the final concentrations are 512μg / mL, 256μg / mL, 128μg / mL, 64μg / mL, 32μg / mL, 16μg / mL, 8μg / mL, 6 replicates per treatment. After culturing in the incubator for 24 hours, add 10 μL of 20% TritonX-100 to the positive control well, blow and mix with a pipette, and continue to culture for 15 minutes; after the culture, centrifuge at 1500 rpm for 10 minutes; after centrifugation, carefully draw 60uL of cells from eac...

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Abstract

The invention discloses a saccharomyces polypeptide derived from saccharomyces cerevisiae var. boulardii, a preparation method for the saccharomyces polypeptide and application of the saccharomyces polypeptide. The saccharomyces polypeptide derived from the saccharomyces cerevisiae var. boulardii is named after FTP-21 and has a complete sequence: SWLSGGFKLKKSVVKRFKLCYCRRFCVF, SEQ ID NO: 1. The preparation method for the polypeptide comprises the step of carrying out chemical artificial synthesis or extracting a fermentation product from the saccharomyces cerevisiae var. boulardii CNCM I-1079. According to use of the polypeptide, the polypeptide is used for selectively killing or inhibiting bacteria including E.coli K 88, E.coli K 12, Salmonella ATCC13076, S.aureus ATCC25923 and C.perfringens ATCC13124; and the polypeptide is used for selectively reserving probiotics including Lactobacillus acidophilus ATCC4356 and Bifidobacterium longum ATCC27533. According to the use of the polypeptide, the polypeptide is used for preparing feed additives for conditioning intestinal florae of dogs and cats and promoting nutrient absorption. According to the use of the polypeptide, the polypeptide is used for preparing immunoregulatory feed additives for improving inflammations of dogs and cats. The saccharomyces polypeptide disclosed by the invention is low in molecular weight, high in net charge, good in stability, high in solubility and easy in digestive absorption and has the characteristics of selective antibacterial activity and high safety.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a yeast polypeptide derived from Saccharomyces bradyi and its preparation method and application. Background technique [0002] Brady's yeast is a tropical yeast isolated from litchi and mangosteen for the first time by French scientist Henri Brad in 1923. It is also called Brady's yeast. kind. Bradiga yeast is one of the few yeast probiotics. Compared with Saccharomyces cerevisiae, Bradiga yeast has a higher survival rate at 37°C, 39°C, 45°C and gastric acid environment for 1 hour, and it has a higher survival rate in bile salt environment The tolerance of Saccharomyces cerevisiae was also significantly higher than that of Saccharomyces cerevisiae, and the antioxidant function of Saccharomyces cerevisiae was also stronger. Brady's yeast has rich metabolites, and the polyamines it secretes help to increase the activity of digestive enzymes, thereby promoting the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/395C12P21/02A23K50/40A23K20/147C12R1/865
CPCC07K14/395C12P21/02A23K50/40A23K20/147Y02P60/87
Inventor 胡汪洋连姝凝刘波静
Owner 杭州吾尾科技有限公司
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