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Dual-enzyme hydrolysis preparation method of anti-tumor polypeptides of spirulina

A spirulina, anti-tumor technology, applied in the biological field, can solve the problem that the anti-tumor peptide is in the primary stage

Active Publication Date: 2015-04-29
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, it has been reported that antioxidant peptides, ACE inhibitory peptides, and bacteriostatic peptides have been successfully extracted from spirulina protein hydrolyzate, but the research on anti-tumor peptides is still in its infancy.

Method used

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  • Dual-enzyme hydrolysis preparation method of anti-tumor polypeptides of spirulina
  • Dual-enzyme hydrolysis preparation method of anti-tumor polypeptides of spirulina
  • Dual-enzyme hydrolysis preparation method of anti-tumor polypeptides of spirulina

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] (1) Take 10 g of spirulina powder, prepare a solution with a concentration of 5% (w / v) with ultrapure water, freeze it in a refrigerator at -20 °C for 4 hours, and then thaw it in a water bath at 37 °C for 2 hours, and freeze it repeatedly Melt 5 times. Homogenize the sample in an ice bath for 2 minutes at 5000rpm for 30s, then 10000rpm for 1min, and finally 5000rpm for 30s. Then in an ice bath, sonicate at a power of 460W for 20min (each sonication is 6 s, with an interval of 9 s). 4°C, 8000r / min, centrifuge for 30 min.

[0031](2) Prepare the spirulina protein obtained in step (1) into a 2% (w / w) protein solution, add alkaline protease Alcalase2.4 to make the enzyme-substrate concentration ratio 4% (w / w), adjust Temperature to 40°C, pH 7.0, enzymatic hydrolysis for 8 h. Then the pH was adjusted to 6.0, and papain was added for enzymolysis according to the ratio of 3% (w / w) of enzyme to substrate for 4 hours. In this process, 0.05 mol / L NaOH and HCl are used to adj...

Embodiment 2

[0036] (1) Take 20 g of spirulina powder, prepare a solution with a concentration of 5% (w / v) with ultrapure water, freeze it in a -20 ℃ refrigerator for 5 hours, and then thaw it in a 37 ℃ water bath for 2.5 hours, and repeat this process Freeze and thaw 4 times. The sample was homogenized in an ice bath for 70s, with a rotation speed of 5000rpm for 20s, then 12000rpm for 30s, and finally 5000rpm for 20s. Then in an ice bath with a power of 500 W ultrasonic 20min (every ultrasonic 4 s, interval 6 s). 4°C, 8000r / min, centrifuge for 40 min.

[0037] (2) Prepare the spirulina protein obtained in step (1) into a 3% (w / w) protein solution, add alkaline protease Alcalase2.4 to make the enzyme-substrate concentration ratio 5% (w / w), adjust Temperature to 50°C, pH 8.0, enzymatic hydrolysis for 6 h. Then adjust the pH to 6.5, and add papain to hydrolyze for 3 hours according to the enzyme-to-substrate ratio of 3.5% (w / w). In this process, 0.05 mol / L NaOH and HCl are used to adjust...

Embodiment 3

[0041] (1) Take 25 g of spirulina powder, prepare a solution with a concentration of 5% (w / v) with ultrapure water, freeze it in a -20 °C refrigerator for 6 hours, and then thaw it in a 37 °C water bath for 3 hours, and freeze it repeatedly Melt 3 times. The samples were homogenized in an ice bath for 150s, first at 4000rpm for 50s, then at 8000rpm for 50s, and finally at 4000rpm for 50s. Then ultrasound was performed at 600 W for 15 min in an ice bath (every ultrasound was 5 s, with an interval of 7 s). 4°C, 8000r / min, centrifuge for 45 min.

[0042] (2) Prepare the spirulina protein obtained in step (1) into a 4% (w / w) protein solution, add alkaline protease Alcalase2.4 to make the enzyme-substrate concentration ratio 5% (w / w), adjust Temperature to 55°C, pH 8.5, enzymatic hydrolysis for 5 hours. Then adjust the pH to 6.5, and add papain to hydrolyze for 3 hours according to the enzyme-substrate ratio of 4% (w / w). In this process, 0.05 mol / L NaOH and HCl are used to adju...

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Abstract

The invention provides a dual-enzyme hydrolysis preparation method of anti-tumor polypeptides of spirulina. The method comprises the following steps: preparing spirulina holoprotein with pure water by means of repeated freezing and thawing, homogenizing and ultrasonic treatment; preparing spirulina polypeptides by means of an alkaline protease Alcalase 2.4 and papain dual-enzyme hydrolysis method; separating and purifying by utilizing an ultrafiltration membrane to acquire spirulina polypeptides having molecular weight ranges of 0-3K, 3-5K, 5-10K and more than 10K. The obtained spirulina polypeptide and monopeptide ingredients can be used for establishing a theoretical basis for development and utilization of anti-tumor medicines and health foods.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a double enzymatic hydrolysis preparation method of spirulina anti-tumor polypeptide. Background technique [0002] Bioactive peptides are special protein fragments that have a positive effect on the function or state of the body and ultimately affect the health of the body. Compared with proteins, the advantages of small-molecule peptide fragments are mainly reflected in: easier to be absorbed and utilized by the human body; high activity, which can exert its unique physiological effects at a small concentration; small molecular weight, easy to modify and transform, can By artificial chemical synthesis etc. Compared with single amino acids, small molecule peptides not only have special physiological activities, but also have incomparable advantages of amino acids in absorption channels and absorption speed. Studies have confirmed that there are special oligopeptide abs...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K7/06C07K1/36C07K1/34C07K1/16A61P35/00
Inventor 张学武王竹君
Owner SOUTH CHINA UNIV OF TECH
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