Preparation method for phycocyanin polypeptide

A technology of phycocyanin and papain, which is applied in the field of protein enzymatic hydrolysis, can solve the problems of singleness and low efficiency of proteolysis, and achieve the effect of improving enzymatic hydrolysis efficiency and solubility

Active Publication Date: 2017-05-24
YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

However, the existing phycocyanin enzymatic hydrolysis methods usually use a single enzymatic hydrolysis technology to enzymatically hydrolyze phycocyanin, which has the disadvantage of low proteolysis efficiency

Method used

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  • Preparation method for phycocyanin polypeptide
  • Preparation method for phycocyanin polypeptide

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preparation example Construction

[0023] Please refer to figure 1 , the preparation method of the phycocyanin polypeptide of one embodiment, comprises the following steps:

[0024] S10, adding water into the phycocyanin, stirring to fully dissolve it, and obtaining a phycocyanin solution.

[0025] Among them, the purity of phycocyanin A 620 / A 280 >2. The solid-liquid ratio of phycocyanin and water can be 1:10-1:50.

[0026] In the operation of S10, the phycocyanin is dissolved by magnetic stirring at a constant temperature, and the temperature may be 50° C. to 55° C. The stirring time may be 20-40 minutes.

[0027] S20. Add papain or neutral protease to the phycocyanin solution, stir evenly, and ultrasonically treat for 30-50 minutes to obtain an enzymatic hydrolysis product.

[0028] Wherein, the ultrasonic treatment can be carried out in a water bath ultrasonic device.

[0029] Wherein, the enzyme activity of papain can be 5×10 5 ~10×10 5 U / g. The mass ratio of papain and phycocyanin may be 1:100-...

Embodiment 1

[0041] (1) Weigh 5g phycocyanin (A 620 / A 280 >2) Put it into a container, add 200mL of distilled water, and stir on a constant temperature magnetic stirrer for 30 minutes to fully dissolve it to obtain a phycocyanin solution;

[0042] (2) The phycocyanin solution obtained in step (1) is adjusted to pH 6.0, and then 1.5g of papain (enzyme activity 8×10 5 U / g), after stirring evenly, ultrasonically treat in a water bath ultrasonic device, the ultrasonic power is 600W, the reaction temperature is 55°C, and the reaction time is 50min, to obtain the enzymatic hydrolysis product;

[0043] (3) Rapidly raise the temperature of the enzymolysis reaction product to 90°C for 10 minutes, cool to 30°C after inactivating the enzyme, centrifuge at 10,000 r / min for 10 minutes, take the supernatant, which is the enzymolysis solution, and the degree of hydrolysis of phycocyanin is 13.68 %;

[0044] (4) The enzymolysis solution obtained in step (3) is subjected to ultrafiltration treatment by...

Embodiment 2

[0047] (1) Weigh 5g phycocyanin (A 620 / A 280>2) Put it into a container, add 50mL of distilled water, and stir on a constant temperature magnetic stirrer until fully dissolved to obtain a phycocyanin solution;

[0048] (2) the pH of the phycocyanin solution obtained in step (1) is adjusted to 5.0, then add papain by 5% of the phycocyanin weight (enzyme activity 10 × 10 5 U / g), after stirring evenly, ultrasonic treatment in a water bath ultrasonic device, the ultrasonic power is 300W, the reaction temperature is 50°C, and the reaction time is 30min, to obtain the enzymatic hydrolysis product;

[0049] (3) Rapidly raise the temperature of the enzymolysis reaction product to 85°C and keep it warm for 15 minutes, cool it down to 30°C after inactivating the enzyme, centrifuge at 10,000r / min for 10 minutes, take the supernatant, which is the enzymolysis solution, and the hydrolysis degree of phycocyanin is measured as 12.97 %;

[0050] (4) using a molecular weight cut-off of 10k...

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Abstract

The invention relates to a preparation method for a phycocyanin polypeptide, which includes the following steps: phycocyanin is added with water for being stirred to be sufficiently dissolved, so that a phycocyanin solution is obtained; the phycocyanin solution is added with papain or neutral protease, uniform stirring is carried out and then ultrasonical treatment is carried out for 30 to 50 minutes, so that an enzymolysis product is obtained; the enzymolysis product is heated to 85 DEG C to 95 DEG C which is kept for 5 to 15 minutes, and after enzyme deactivation, cooling and centrifugation, supernatant, i.e. enzymolysis liquid, is extracted; ultrafiltration treatment is carried out on the enzymolysis liquid, so that an ultrafiltrate is obtained; the ultrafiltrate is concentrated under vacuum, and after spray-drying treatment, the phycocyanin polypeptide is obtained. The preparation method for the phycocyanin polypeptide adopts ultrasound-assisted enzymolysis, ultrasonic waves have a cavitation effect, produced air bubbles are squeezed to break to instantly produce extremely strong mechanical shear force, so that protein is degraded or protein conformation is changed, consequently, more hydrophilic groups are promoted to expose, the solubility of phycocyanin in water is increased, the binding between enzyme and phycocyanin substrate is benefited, and the efficiency of enzymolysis of phycocyanin can be remarkably increased.

Description

technical field [0001] The invention relates to the technical field of protein enzymatic hydrolysis, in particular to a preparation method of phycocyanin polypeptide. Background technique [0002] Phycocyanin (also known as PC) is a photosynthetic accessory pigment present in the cells of red algae and cyanobacteria, which can efficiently capture light energy. In recent years, studies have found that phycocyanin has various biological activities such as anti-oxidation, anti-cancer, anti-inflammation and neuroprotection, and its research has become a hot spot in the study of natural marine medicines. my country's research on phycocyanin began in the 1970s. After more than 30 years of research and development, although great progress has been made, most of them are at the laboratory level. At the same time, there are not many functional foods based on phycocyanin, and there is still a gap in the research and development of medicines. Enzymatic hydrolysis can improve the func...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K14/795C07K1/34
CPCC12P21/06C07K14/795Y02P20/10
Inventor 李文军唐志红秦松
Owner YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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