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Spirulina polypeptide P1 with bacteriostatic activity, and application thereof

A technology of spirulina polypeptide and antibacterial activity, applied in the field of protein engineering, can solve problems such as unfavorable processing, unfavorable digestion and absorption, high viscosity, etc., and achieve the effect of broad application prospects.

Active Publication Date: 2015-01-28
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, protein is a macromolecular substance, and its viscosity is relatively large after it swells with water, which is not conducive to processing, and it is also not conducive to the digestion and absorption of it by the human body

Method used

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  • Spirulina polypeptide P1 with bacteriostatic activity, and application thereof
  • Spirulina polypeptide P1 with bacteriostatic activity, and application thereof
  • Spirulina polypeptide P1 with bacteriostatic activity, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] The extraction of embodiment 1 spirulina protein

[0018] (1) Dissolve an appropriate amount of Spirulina platensis dry powder in distilled water to make a Spirulina suspension, and use ultrasound combined with repeated freezing and thawing to break the wall.

[0019] (2) Centrifuge the spirulina liquid obtained in step (1) (-4°C, 8000r / min for 20min) to collect the supernatant, and use 50% saturated NH4SO4 for salting-out purification.

[0020] (3) Refrigerate and centrifuge the spirulina liquid obtained after salting out in step (2) (10000r / min for 20min), collect the precipitate, dissolve it in a phosphate buffer solution with a concentration of 0.005M and a pH of 6.86, and dialyze at 4°C , The end point of dialysis was detected with BaCl2. After desalination, vacuum freeze-drying is carried out to obtain the spirulina protein.

Embodiment 2

[0021] The extraction of embodiment 2 spirulina mixed polypeptides

[0022] (1) Carry out enzymatic hydrolysis of the spirulina protein prepared in Example 1. First, use alkaline protease to hydrolyze the spirulina protein. The enzymatic hydrolysis conditions are: the amount of enzyme added is 4300U / g, the temperature is 55°C, and the pH is 7.0. 160min; after hydrolysis, inactivate the enzyme in a water bath at 85°C for 15-20min.

[0023] (2) The hydrolyzate of step (1) was further hydrolyzed with papain, the enzymolysis conditions were: enzyme-to-substrate ratio 4.5%, temperature 60°C, pH 6.5, enzymolysis 210min. After hydrolysis, inactivate the enzyme in a water bath at 80°C for 15-20 minutes.

[0024] (3) Centrifuge the hydrolyzate obtained in step (2) at 4°C, 5000r / min for 20min, collect the supernatant, freeze-dry to obtain the Spirulina polypeptide.

Embodiment 3

[0025] Example 3 Separation, purification and artificial synthesis of bacteriostatic spirulina polypeptide

[0026] (1) The spirulina mixed polypeptide prepared in Example 2 was subjected to antibacterial experiments, using Escherichia coli and Staphylococcus aureus as the experimental strains to confirm that it has antibacterial activity.

[0027] (2) Separate and purify the mixed polypeptide with antibacterial properties in step (1) using molecular sieves, and use Sephadex G-25 column chromatography to obtain 4 components, and conduct antibacterial experiments on each component , the results showed that component 2 had antibacterial activity.

[0028] (3) The component 2 obtained in step (2) was separated and purified by reverse-phase high-performance liquid chromatography (Zorbax SB-C18 column 4.6mm×250mm), and 4 groups of peaks were obtained. After collection, antibacterial experiments were carried out to obtain Peak 1 has antibacterial activity.

[0029] (4) The peak 1 ...

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Abstract

The invention provides a novel spirulina polypeptide P1 with bacteriostatic activity. An amino acid sequence of the spirulina polypeptide P1 is shown as Seq ID No.1. The spirulina polypeptide P1 with bacteriostatic activity is extracted from spirulina for the first time. Experiments demonstrate that the spirulina polypeptide P1 can effectively suppress gram-positive bacteria and gram-negative bacteria, has no toxic or side effect, can be used as a broad-spectrum antibacterial agent or a bacteria preservative, and has wide application prospects.

Description

technical field [0001] The invention relates to the field of protein engineering, in particular to a spirulina polypeptide P1 with antibacterial activity and its application. Background technique [0002] Spirulina (Spirulina) is a class of single-celled organisms belonging to the cyanophyta Oscillatoriaceae. A large number of studies have shown that spirulina is very rich in nutrients, the protein content is as high as 50%-70%, and the proportion of amino acids is very ideal. It is an excellent source of protein. However, protein is a macromolecular substance, which has a high viscosity after being swollen with water, which is not conducive to processing, and is also not conducive to the digestion and absorption of it by the human body. The hydrolysis of spirulina protein is beneficial to improve the solubility and utilization rate of protein. Spirulina protein is hydrolyzed to form polypeptides and small molecular peptides, which can be quickly absorbed by the human body...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/195A61K38/16A61P31/04A23L3/3526
CPCA23L3/3526A23V2002/00A61K38/00C07K14/195A23V2200/10A23V2250/55
Inventor 李博生孙宜君
Owner BEIJING FORESTRY UNIVERSITY
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